MUTATED NON-PRIMATE LENTIVIRAL ENV PROTEINS AND THEIR USE AS DRUGS

Information

  • Patent Application
  • 20160129107
  • Publication Number
    20160129107
  • Date Filed
    June 06, 2014
    10 years ago
  • Date Published
    May 12, 2016
    8 years ago
Abstract
A pharmaceutical composition includes, as active substance a mutated non-primate lentiviral Env protein having decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties, or a variant of the mutated lentiviral Env protein, or a fragment of the above proteins, in association with a pharmaceutically acceptable carrier.
Description

The present invention relates to mutated non-primate lentiviral ENV proteins and their use as drugs.


Lentivirus is a genus of slow viruses of the Retroviridae family, characterized by a long incubation period. Lentiviruses have the unique ability among retroviruses of being able to integrate into the DNA of the host and replicate in non-dividing cells. They are in most cases pathogenic, infection resulting in a severe immunodeficiency syndrome (AIDS).


Lentiviruses can be found in primates including humans (HIV1, HIV2) and monkeys (SIV), as well as in other animals.


As examples of animal lentiviruses, one may cite feline lentiviruses such as Feline Imunodeficiency Virus (FIV) which can be found in cats, as well as in other felidae (e.g. lion, cougar).


Other examples of animal lentiviruses are bovine immunodeficiency virus (BIV), Jembrana disease virus (JDV, BIV from Indonesia), equine infectious anaemia virus (EIAV), caprine arthritis encephalitis virus (CAEV), Visna/Maedi virus (VMV) and ovine Visna/Maedi virus (OMVV).


Feline imunodeficiency virus (FIV) causes an AIDS-like syndrome in the domestic cat, with marked similarity to AIDS caused by human immunodeficiency virus (HIV) in humans. FIV in cats typically manifests itself as a transient acute-phase syndrome, characterized by febrile episodes, lymphadenopathy, neutropenia, and weight loss. This initial phase is followed by a protracted asymptomatic period with progressive loss of CD4+T lymphocytes and a terminal AIDS phase characterized by succumbing to opportunistic infections. Variability exists among FIV strains, not only in sequence relatedness but also in pathogenicity in vivo. FIV has been reported worldwide with a prevalence rate ranging from 1 to 28%. It affects 2-4% of cats in the US.


Following criteria similar to those for HIV subgroups distinction, FIV has been classified into subgroups according to the envelop protein (Env) diversity. Sodora et al. defined the original Petaluma strain as a prototype subgroup A and the divergent Japanese TM2 strain as prototype subgroup B. A majority of FIV sequences obtained worldwide were categorized in the A and B subgroups. A subtype C was also defined and is the least prevalent FIV subgroup, as well as two other subtypes D and E.


As in the case of HIV, the development of an effective vaccine against FIV is difficult because of the variations of the virus strains, most probably associated with the high intrinsic mutation rate of its replicative machinery. A dual-subtype vaccine for FIV released in 2002 called Fel-O-Vax was shown to have some protective effect against subtypes A and B FIV, but a more effective vaccine is still needed for an efficient protection of cats.


A key feature that could result in higher efficacy of a FIV dedicated vaccine is to increase the immunogenicity of the FIV antigens that are introduced in the vaccine. In this respect, it is essential to ascertain that the antigens do not carry an immunosuppressive activity, which would severely decrease the immunogenicity of the vaccine. In fact, most vaccines against retroviruses contain the Env protein as it is the first exposed viral protein to be <<seen>> by the immune system of an infected animal, and it is important for a vaccinated animal to be able to mount an efficient line of defence against this protein. Yet, it has been shown in the case of the non-lentiviral retroviruses—including oncoretroviruses such as the feline FeLV or the human HTLV retroviruses—that their Env protein carries an immunosuppressive activity which drastically decreases the immunogenicity of the corresponding proteins, and severely impairs the efficacy of a vaccine expressing the Env protein.


Consequently for a vaccine, there is a need to provide proteins as antigens having lost, or substantially lost, their immunosuppressive functions, in order to generate an efficient response. This will enable the animals once infected by the virus to allow the immune system to destroy the infected cells and prevent/cure the infection.


One aim of the invention is to provide new mutated Env proteins devoid of immunosuppressive properties.


Another aim of the invention is to provide new mutated animal Env proteins.


Another aim of the invention is to provide a new pharmaceutical composition efficient for preventing/treating lentiviral infection.


Another aim of the invention is to provide an efficient vaccine against animal lentiviruses, in particular feline lentiviruses.


The invention relates to a pharmaceutical composition comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,


said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein,
    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K or deleted, and/or
    • X3 is any amino acid different from F or deleted, and/or
    • X4 is any amino acid different from L or deleted, and/or
    • X5 is any amino acid different from Y or deleted,


      in association with a pharmaceutically acceptable carrier.


In particular, the invention relates to a pharmaceutical composition comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


in particular said fragment of said isolated mutated feline lentiviral ENV protein comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,


said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein,
    • X1 is A, F, G, L or R, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is A, F, G, L or R, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is A, G, L or R, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is A, F, G or R, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is A, F, G, L or R, and X1, X2, X3 and X4 are any amino acid,


      in association with a pharmaceutically acceptable carrier.


The present application is based on the unexpected observation made by the Inventors that some specific amino acids of the immunosuppressive domain (ISU) of a lentiviral Env protein can be mutated conferring to said lentiviral Env protein decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties, while retaining its antigenicity, the three-dimensional structure of the immunosuppressive domain, and its expression at the plasma membrane.


The expression “having decreased immunosuppressive properties” means that the mutated Env protein has lost at least 50% of its immunosuppressive activity in comparison to the immunosuppressive activity of the wild type Env protein.


The expression “having substantially no immunosuppressive properties” means that the mutated Env protein has lost at least 75% of its immunosuppressive activity in comparison to the immunosuppressive activity of the wild type Env protein.


The expression “having no immunosuppressive properties” means that the mutated Env protein has lost 100% of its immunosuppressive activity in comparison to the immunosuppressive activity of the wild type Env protein.


Moreover, the mutated lentiviral Env protein according to the invention has retained a part or the totality of its fusogenic activity.


In the invention, the expression “mutated feline lentiviral ENV proteins” means that the ENV proteins derive from the expression of an env gene of a lentivirus of a feline.


Feline lentiviruses according to the invention encompass the FIV which can be found in cats, as well as in other felidae (e.g. lion, cougar).


Because of the natural variability of feline lentiviruses, the “mutated Env protein”, as defined in the invention, encompasses two meanings.


According to the first meaning, the said “mutated ENV protein” is the unnatural result of the intervention of human beings.


According to the second meaning, the mutated Env protein also encompasses naturally occurring variants for which up to now the non immunosuppressive properties remain unknown.


This second meaning takes into consideration the natural variability of FIV variants inside a same infected animal, wherein the said “mutated Env protein” might be non immunosuppressive but its property is undetectable because an FIV infected animal carries many FIV variants, the majority of which is immunosuppressive.


The following protein (Accession No. P16090) corresponds to the wild type sequence of the Env protein of FIV (Petaluma strain). In the invention, it is considered as a reference sequence of the wild type ENV protein.









wild type FIV Env


(Petaluma strain)







SEQ ID NO: 4







MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGVNPFRVPGI





TEKEKQNYCNILQPKLQDLRNEIQEVKLEEGNAGKFRRARFLRYSDE





RVLSLVHAFIGYCIYLGNRNKLGSLRHDIDIEAPQEECYNNREKGTT





DNIKYGRRCCLGTVTLYLILFTGVIVYSQTAGAQVVWRLPPLVVPVE





ESEIIFWDCWAPEEPACQDFLGAMIHLKAKTNISIREGPTLGNWARE





IWATLFKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIVPDY





QCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLSYCTDPLQIPLI





NYTFGPNQTCMWNTSQIQDPEIPKCGWWNQMAYYNSCKWEEAKVKFH





CQRTQSQPGSWFRAISSWKQRNRWEWRPDFKSKKVKISLPCNSTKNL





TFAMRSSGDYGEVTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDT





SLIDTCGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDLIVHF





NMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTNSSSSYSGTKMACP





SNRGILRNWYNPVAGLRQSLEQYQVVKQPDYLLVPEEVMEYKPRRKR





AAIHVMLALATVLSIAGAGTGATAIGMVTQYHQVLATHQEAIEKVTG





ALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQNQFF





CKIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEIIMD





IEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLKGLLGGILGIGLGVL





LLILCLPTLVDCIRNCIHKILGYTVIAMPEVEGEEIQPQMELRRNGR





QCGMSEKEEE






Variants of the FIV mutated Env proteins according to the invention have at least 70%, preferably at least 80%, more preferably at least 90% of identity with the wild type amino acid sequence of the FIV Env protein, and comprise the mutations as described above, and harbour a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity.


A region of the feline lentiviral Env protein containing the amino acids which may be mutated according to the invention can be delimited by the sequence SEQ ID NO: 2:









(SEQ ID NO: 2)









G-L-[K/T/R]-[V/I]-[E/R]-A-[I/M/T/L]-E-K-[F/P]-






[L/V/I]-Y-T-A-[F/L]-A-M.






SEQ ID NO: 2 corresponds to the consensus sequence of a specific region of the ISU domain. In the invention, the ISU domain comprises SEQ ID NO: 2.


When applying the mutations as defined above, the ISU domain of the Env protein loses partially or totally its immunosuppressive properties.


Examples of fragments of the wild type ISU domains of FIV:












Examples of fragments of the wild type


ISU domains of FIV:















(SEQ ID NO: 6)


AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKIPLELWTR





(SEQ ID NO: 7)


AVEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKIPPELWTR





(SEQ ID NO: 8)


AIEKVTEALKITNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKVPPELWRR





(SEQ ID NO: 9)


AIEKVTEALKINNLRLVTLEHQVLVIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCKVPLELWRR





(SEQ ID NO: 10)


AIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKVPSALWER





(SEQ ID NO: 11)


AIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKIPPGLWTR





(SEQ ID NO: 12)


AIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKVPPQLWKR





(SEQ ID NO: 13)


AIEKVTEALKVNNLRLITLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKVPPELWKR





(SEQ ID NO: 14)


TIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKVPPELWKR





(SEQ ID NO: 15)


AIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCNQ





NQFFCKVPPVLWER





(SEQ ID NO: 16)


TIEKITEALKVNNLRLVTLEHQVLVIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCKVPPELWQR





(SEQ ID NO: 17)


AIDQITEALKINNLRLVTLEHQVLVIGLKVEAIEKFIYTAFAMQELGCNQ





NQFFCKIPPELWIR





(SEQ ID NO: 18)


ALDKITEALKINNLRLITLEHQVLVIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCKIPPSLWSM





(SEQ ID NO: 19)


ALDKITEALKINNLRLVTLEHQVLVIGLKVEATEKFLYTAFAMQELGCNQ





NQFFCKIPCELWMR





(SEQ ID NO: 20)


ALEKITEALKINNLRLVTLEHQVLMIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCKIPLNLWTM





(SEQ ID NO: 21)


ALDKITEALKINNLRLITLVHQVLVIGLKVRAIEKPLYTAFAMQELGCNQ





NQFFCKIPPSLWSM





(SEQ ID NO: 22)


ALEKITEALKINNLRLITLEHQVLVIGLRVEAIEKFLYTAFAMQELGCNQ





NQFFCKIPPSLWSM





(SEQ ID NO: 23)


ALEKITEALKINNLRLITLEHQVLVIGLKVEAIEKFLYTAFAMQELGCHQ





NQFFCKIPPSLWSM





(SEQ ID NO: 24)


ALDKITEALKINNLRLITLEHQVLVIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCKIPPSLWSM





(SEQ ID NO: 25)


ALDKITQALKINNLRLITLEHQVLVIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCKIPPSLWSM





(SEQ ID NO: 26)


ALDKITEALKINNLRLITLEHQVLVIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCKIPLNLWNM





(SEQ ID NO: 27)


ALDKITEALKINNLRLVTLEHQMLVIGLKVEAIEKFLYTAFAMQELGCNQ





NQFFCEIPKELWLR









The localization of an ISU domain can be determined in all Env proteins of viruses as described in Benit et al. 2001, Journal of Virology, Vol. 75, No. 23, p. 11709-11719. In a broad meaning, the ISU domain is defined by its structure and its localization, irrespective of the fact that it possesses or not an immunosuppressive activity.


In the invention, the ISU domain refers to a specific domain in which a mutation can affect the immunosuppressive property of the ENV protein.


As an example of a mutated feline lentiviral Env protein, SEQ ID NO: 5 corresponds to the sequence of a mutated Env protein of the FIV Petaluma strain. In the invention, it is considered as a reference sequence of the mutated Env protein.


More specifically, SEQ ID NO: 5 corresponds to the SEQ ID NO: 4 in which the amino acid residue F in position 5 (X3) of the sequence A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A (SEQ ID NO: 1) has been substituted by R.











mutated FIV Env







SEQ ID NO: 5









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGVNPFRV






PGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGNAGKFRRARF






LRYSDERVLSLVHAFIGYCIYLGNRNKLGSLRHDIDIEAPQEEC






YNNREKGTTDNIKYGRRCCLGTVTLYLILFTGVIVYSQTAGAQV






VWRLPPLVVPVEESEIIFWDCWAPEEPACQDFLGAMIHLKAKTN






ISIREGPTLGNWAREIWATLFKKATRQCRRGRIWKRWNETITGP






SGCANNTCYNVSVIVPDYQCYLDRVDTWLQGKINISLCLTGGKM






LYNKVTKQLSYCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIP






KCGWWNQMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGEVTGAW






IEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDTCGNTPNVSG






ANPVDCTMYSNKMYNCSLQNGFTMKVDDLIVHFNMTKAVEMYNI






AGNWSCTSDLPSSWGYMNCNCTNSSSSYSGTKMACPSNRGILRN






WYNPVAGLRQSLEQYQVVKQPDYLLVPEEVMEYKPRRKRAAIHV






MLALATVLSIAGAGTGATAIGMVTQYHQVLATHQEAIEKVTGAL






KINNLRLVTLEHQVLVIGLKVEAMEKRLYTAFAMQELGCNQNQF






FCKIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYE






IIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLKGLLGGIL






GIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAMPEVEGEEIQ






PQMELRRNGRQCGMSEKEEE






The invention also encompasses the variants of the “mutated feline lentiviral Env protein”, harbouring the above mentioned mutations, and conferring a decrease or a lack of immunosuppressive properties to said variant.


Variants of the FIV mutated Env proteins according to the invention have at least 70%, preferably at least 80%, more preferably at least 90% of identity with the reference mutated sequence of FIV env protein (SEQ ID NO: 5), and comprise the mutations as described above, and harbour a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity.


Variants of the FIV mutated Env proteins according to the invention have at least 90% of identity with the transmembrane region of SEQ ID NO 5


According to a particular embodiment of the invention, variants of the FIV mutated Env proteins according to the invention have a transmembrane region which has at least 90% of identity with the transmembrane region of the FIV mutated Env protein, said variants comprising a mutated immunosuppressive domain (ISU) containing the sequence SEQ ID NO: 1.


According to another particular embodiment of the invention, variants of the FIV mutated Env proteins according to the invention have a 64 amino acid sequence, corresponding to a fragment of the ISU domain, which has at least 95% of identity with a 64 amino acid sequence of the FIV mutated Env protein comprising the sequence SEQ ID NO: 1.


In a particular embodiment, the invention relates to a pharmaceutical composition as defined above comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof, said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,


said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 3)


[V/I]-[E/R]-A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A-[F/L]-





A-M,







wherein,
    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K or deleted, and/or
    • X3 is any amino acid different from F or deleted, and/or
    • X4 is any amino acid different from L or deleted, and/or
    • X5 is any amino acid different from Y or deleted,


      in particular, wherein,
    • X1 is A, F, G, L or R, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is A, F, G, L or R, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is A, G, L or R, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is A, F, G or R, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is A, F, G, L or R, and X1, X2, X3 and X4 are any amino acid,


      in association with a pharmaceutically acceptable carrier.


SEQ ID NO: 3 (14 amino acid long) is a consensus sequence containing SEQ ID NO: 1 (9 amino acid long).


The invention relates to a pharmaceutical composition as defined above comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein, said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,


said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein,
    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K or deleted, and/or
    • X3 is any amino acid different from F or deleted, and/or
    • X4 is any amino acid different from L or deleted, and/or
    • X5 is any amino acid different from Y or deleted,


      in particular, wherein,
    • X1 is A, F, G, L or R, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is A, F, G, L or R, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is A, G, L or R, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is A, F, G or R, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is A, F, G, L or R, and X1, X2, X3 and X5 are any amino acid,


      in association with a pharmaceutically acceptable carrier,


said decrease, substantial absence or absence of immunosuppressive activity of the above mentioned mutated feline lentiviral ENV protein or of the above defined fragment being liable to be assessed by the fact that in an in vivo assay involving engrafted tumor cells rejection, in animals excluding human beings,


said tumor cells being transduced either so as to express said mutated ENV protein or said fragment (mutated ENV tumor cells),


or said tumor cells being transduced so as to express said wild type ENV protein or a fragment thereof (wild type ENV tumor cells),


or said tumor cells being not transduced (normal tumor cells), the following ratio:


immunosuppression index of said mutated ENV protein or of said fragment (imutated env)/immunosuppression index of wild type ENV protein (iwild type env) is less than 0.5, or even less than 0.25,


imutated env being defined by: (maximum area reached by mutated ENV tumor cells−maximum area reached by normal tumor cells)/(maximum area reached by normal tumor cells), and


iwild type env being defined by: (maximum area reached by wild type ENV tumor cells−maximum area reached by normal tumor cells)/(maximum area reached by normal tumor cells).


The immunosuppressive property of the ENV protein is preferably measured using in vivo procedures, which are representative of the physiological environment.


The immunosuppressive property of the mutated ENV proteins according to the invention is measured according to an in vivo procedure to assay the immunosuppressive activity of a ENV protein disclosed previously [Mangeney and Heidmann Proc Natl Acad Sci USA 1998; 95: 14920-14925; Mangeney et al. Proc Natl Acad Sci USA, 2007, 104(51):20534-9].


As a physiological test, this in vivo procedure is performed using ENV proteins, or fragment thereof, which are not associated to another component or carrier proteins, such as BSA.


Briefly, a wild-type (wild type lentiviral ENV protein) or modified nucleic acid expressing the protein to be tested (mutated lentiviral ENV protein) is transduced in tumour cell lines such as MCA 205 or C18.1 cell lines by known transduction methods. The tumour cells expressing the protein to be tested are then injected especially subcutaneous (s.c.) injection to a host, generally mice. Following said injection, the establishment of tumour or, to the contrary, its rejection, is determined and the tumour area is measured. Tumour establishment is determined by palpation and tumour area (mm2) is determined by measuring perpendicular tumour diameters. Immunosuppression index is defined as i=(Senv−Snone)/Snone, wherein Senv is the maximum area reached by a tumour expressing an envelope protein and Snone is the maximum area reached by a tumour not expressing ENV protein (negative control). According to an embodiment of the invention, the above defined ratio relative to the immunosuppression index (imutated env/iwild type env) can be less than 0.25, and can even have a negative value (see FIG. 3).


In vitro assay could be carried out, using high doses of synthetic peptides but they are indirect and less convincing, since the expression “immunosuppressive” is relevant when applied to animals possessing a complete immune system and not to cell lines. An additional difficulty for the functional characterization of an ISU domain relies on the fact that the ISU carried by the retroviral Env proteins is a highly structured proteic domain, with trimer formation within the complete Env proteins (Caffrey M., Biochimica et Biophysica Acta, 1536:116-122, 2001; Caffrey et al., The EMBO Journal, Vol. 17, No. 16, p. 4572-4584, 1998). Such structures are not naturally formed with ISU peptides of limited length, and this is most probably why most studies carried out with peptides provide irrelevant results and/or are dependent on specific coupling of the peptides to carrier proteins (such as BSA, e.g. Denner et al., Current Science, AIDS 1994, 8:1063-1072).


As mentioned above, the Env proteins according to the invention are mutated. This mutation is made in vitro. Thus, the mutated Env proteins according to the invention are isolated, and do not correspond to naturally occurring counterpart.


As mentioned above, the lentiviral mutated Env proteins have decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties. This means that the mutated Env proteins according to the invention have no, or have reduced immunosuppressive properties with respect to the natural non mutated Env proteins from a virus of the same species. For instance, a mutated FIV Env protein according to the invention has reduced immunosuppressive properties with respect to the wild type FIV Env protein.


In the invention, the terms “a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity” means that the mutated Env proteins according to the invention have an immunosuppressive index less than about 50 or 25% of that of the wild type Env protein [Mangeney and Heidmann Proc Natl Acad Sci USA 1998; 95: 14920-14925; Mangeney et al. Proc Natl Acad Sci USA, 2007, 104(51):20534-9].


In the invention, structures responsible for the antigenicity of the mutated lentiviral ENV protein are essentially preserved.


As intended herein, the expression “structures responsible for antigenicity” relates to structures of the protein which are liable to interact with components of the immune system such as antibodies or membrane receptors of immune cells, in particular T cells.


The mutation(s) within the immunosuppressive domain of the lentiviral Env proteins is (are) sufficient to decrease the immunosuppressive activity of the mutated lentiviral Env protein with respect to the corresponding wild type Env. However, it might be advantageous that another amino acid be also mutated because it ensures that the structure of the mutated Env protein is essentially conserved with respect to the corresponding wild type Env protein.


The mutated lentiviral Env protein has substantially retained the structure, especially the antigenic structure, e.g., immunogenic determinants, of the original determined lentiviral Env protein, i.e. the wild type non mutated lentiviral Env protein.


These properties can be evaluated by testing the ability of the Env proteins to be normally expressed at the cell membrane under conditions which preserve the surface and transmembrane subunits (SU:TM) interactions and recognition by specific anti-Env antibodies, and by measuring the functional fusogenic activity of said mutated lentiviral Env with respect to the same properties in the wild type non mutated lentiviral Env protein (see examples).


Generally speaking, the mutated ENV protein involved in the present invention has an average length of about 750 to about 1000 amino acids.


The invention encompasses fragments of the mutated ENV protein as defined above, provided that said fragment:

    • comprises at least the sequence SEQ ID NO: 1, as defined above,
    • comprises at least 40 amino acids, preferably comprises at least 50 amino acids,
    • has decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties, as defined above,
    • preferably, comprises the extracellular parts of the ENV protein,
    • retains the structure of the ENV protein from which it derives,
    • harbours the same epitopes as the corresponding fragment in the wild type ENV protein.


According to a particular embodiment, the fragment of the mutated ENV protein of the invention can comprise about 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 amino acids. These values are given only in an illustrative way, as the man skilled in the art will understand that the fragment can comprise any number of amino acids comprised from about 40 to about 950 amino acids.


Advantageously, the fragments according to the invention are such that, while retaining the antigenic structure of the full length mutated ENV protein, and thus of the wild type ENV protein, they have lost major antigenic regions that are responsible for antigenicity in another region than the region corresponding to the immunosuppressive domain, because said regions could be detrimental for targeting an immune response against the immunosuppressive domain.


The invention also relates to a pharmaceutical composition wherein the active substance is an isolated non naturally occurring feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, as above defined, or fragments thereof.


In a particular embodiment, the invention relates to a pharmaceutical composition as defined above wherein:

    • X1 is any amino acid different from E or deleted, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is any amino acid different from F or deleted, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is any amino acid different from L or deleted, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is any amino acid different from Y or deleted, and X1, X2, X3 and X4 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, and X3, X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, and X2, X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X4 is any amino acid different from L or deleted, and X2, X3 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X5 is any amino acid different from Y or deleted, and X2, X3 and X4 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, and X1, X4 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, and X2, X4 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X5 is any amino acid different from Y or deleted, and X2, X3 and X4 are any amino acid, or
    • X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X2, X4 and X5 are any amino acid, or
    • X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X2, X3 and X4 are any amino acid, or
    • X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X2, X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, and X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, and X3 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X5 is any amino acid different from Y or deleted, and X3 and X4 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X2 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X2 and X4 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X2 and X3 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X1 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X1 and X4 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X1 and X3 are any amino acid, or
    • X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X1 and X2 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X5 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X4 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X3 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X2 is any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted and X1 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted and X5 is any amino acid different from Y or deleted.


In a particular embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated immunosuppressive domain contains the amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 3, wherein:

    • X1 is A, F, G, L or R, X2 is A, F, G, L or R, and X3, X4 and X5 are any amino acid, or
    • X1 is A, F, G, L or R, X3 is A, G, L or R, and X2, X4 and X5 are any amino acid, or
    • X1 is A, F, G, L or R, X4 is A, F, G or R, and X2, X3 and X5 are any amino acid, or
    • X1 is A, F, G, L or R, X5 is A, F, G, L or R, and X2, X3 and X4 are any amino acid, or
    • X2 is A, F, G, L or R, X3 is A, G, L or R, and X1, X4 and X5 are any amino acid, or
    • X2 is A, F, G, L or R, X4 is A, F, G or R, and X1, X3 and X5 are any amino acid, or
    • X2 is A, F, G, L or R, X5 is A, F, G, L or R, and X1, X3 and X4 are any amino acid, or
    • X3 is A, G, L or R, X4 is A, F, G or R, and X1, X2 and X5 are any amino acid, or
    • X3 is A, G, L or R, X5 is A, F, G, L or R, and X1, X2 and X4 are any amino acid, or
    • X4 is A, F, G or R, X5 is A, F, G, L or R, and X1, X2 and X3 are any amino acid.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:


X1 is any amino acid different from E or deleted, and/or

    • X2 is any amino acid different from K, or deleted, and/or
    • X3 is any amino acid different from F, P or deleted, and/or
    • X4 is any amino acid different from L, V, I or deleted, and/or
    • X5 is any amino acid different from Y or deleted.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K, A, N, Y, R or deleted, and/or
    • X3 is any amino acid different from F, P or deleted, and/or
    • X4 is any amino acid different from L, V, I or deleted, and/or
    • X5 is any amino acid different from Y, K, E, Q, R, A or deleted.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X1 is R, G, L, A, F or deleted, and/or
    • X2 is R, G, L, A, F or deleted, and/or
    • X3 is R, G, L, A, or deleted, and/or
    • X4 is R, G, A, F or deleted, and/or
    • X5 is R, G, L, A, F or deleted.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X1 is R, G, L, A, F or deleted, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is R, G, L, A, F or deleted, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is R, G, L, A or deleted, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is R, G, A, F or deleted, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is R, G, L, A, F or deleted, and X1, X2, X3 and X4 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X2 is R, G, L, A, F or deleted, and X3, X4 and X5 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, and X2, X4 and X5 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X4 is R, G, A, F or deleted, and X2, X3 and X5 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X5 is R, G, L, A, F or deleted, and X2, X3 and X4 are any amino acid, or
    • X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, and X1, X4 and X5 are any amino acid, or
    • X2 is R, G, L, A, F or deleted, X4 is R, G, A, F or deleted, and X2, X4 and X5 are any amino acid, or
    • X2 is R, G, L, A, F or deleted, X5 is R, G, L, A, F or deleted, and X2, X3 and X4 are any amino acid, or
    • X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, and X2, X4 and X5 are any amino acid, or
    • X3 is R, G, L, A or deleted, X5 is R, G, L, A, F or deleted, and X2, X3 and X4 are any amino acid, or
    • X4 is R, G, A, F or deleted, X5 is R, G, L, A, F or deleted and X2, X4 and X5 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, and X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is R, G, L, A, F or deleted, X4 is R, G, A, F or deleted, and X3 and X5 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X2 is R, G, L, A, F or deleted, X5 is R, G, L, A, F or deleted and X3 and X4 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, and X2 and X5 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X5 is R, G, L, A, F or deleted, and X2 and X4 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X4 is R, G, A, F or deleted, X5 is R, G, L, A, F or deleted, and X2 and X3 are any amino acid, or
    • X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, and X1 and X5 are any amino acid, or
    • X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X5 is R, G, L, A, F or deleted, and X1 and X4 are any amino acid, or
    • X2 is R, G, L, A, F or deleted, X4 is R, G, A, F or deleted, X5 is R, G, L, A, F or deleted, and X1 and X3 are any amino acid, or
    • X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, X5 is R, G, L, A, F or deleted, and X1 and X2 are any amino acid, or
    • X1 is R, G, L, A, F or deleted, X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, and X5 is any amino acid, or
    • X1 is R, G, L, A, F or deleted, X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X5 is R, G, L, A, F or deleted, and X4 is any amino acid, or
    • X1 is R, G, L, A, F or deleted, X2 is R, G, L, A, F or deleted, X4 is R, G, A, F or deleted, X5 is R, G, L, A, F or deleted, and X3 is any amino acid, or
    • X1 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, X5 is R, G, L, A, F or deleted, and X2 is any amino acid, or
    • X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, X5 is R, G, L, A, F or deleted, and X1 is any amino acid, or
    • X1 is R, G, L, A, F or deleted, X2 is R, G, L, A, F or deleted, X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted and X5 is R, G, L, A, F or deleted.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated immunosuppressive domain contains the amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 3, wherein:

    • X1 is A, G or R, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is A, G or R, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is A, G or R, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is A, G or R, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is A, G or R, and X1, X2, X3 and X4 are any amino acid, in particular:
    • X1 is R, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is R, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is R, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is R, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is R, and X1, X2, X3 and X4 are any amino acid.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X1 is R, G, L, A, F or deleted, and/or
    • X2 is G, L, F or deleted, and/or
    • X3 is R, G, L, A, or deleted, and/or
    • X4 is R, G, A, F or deleted, and/or
    • X5 is G, L, F or deleted.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X3 is A, G, L, R, C, D, E, H, K, M, N, Q, S, T, V, W, Y or deleted, X4 is C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X4 is A, F, G, R, C, D, E, H, K, M, N, P, Q, S, T, W, Y or deleted, X3 is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X3 is A, G, L, R, C, D, E, H, K, M, N, Q, S, T, V, W, Y or deleted, X4 is A, F, G, R, C, D, E, H, K, M, N, P, Q, S, T, W, Y or deleted, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, 5, T, V, W or Y.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X3 is R, G, L, A or deleted, X4 is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X4 is R, G, L, A, F or deleted, X3 is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, 5, T, V, W or Y.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X3 is R, A or deleted, X4 is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X4 is R, A or deleted, X3 is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X3 is R, A or deleted, X4 is R, A or deleted, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, 5, T, V, W or Y.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated immunosuppressive domain contains the amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 3, wherein:

    • X1 is A, G or R, X2 is A, G or R, and X3, X4 and X5 are any amino acid, or
    • X1 is A, G or R, X3 is A, G or R, and X2, X4 and X5 are any amino acid, or
    • X1 is A, G or R, X4 is A, G or R, and X2, X3 and X5 are any amino acid, or
    • X1 is A, G or R, X5 is A, G or R, and X2, X3 and X4 are any amino acid, or
    • X2 is A, G or R, X3 is A, G or R, and X1, X4 and X5 are any amino acid, or
    • X2 is A, G or R, X4 is A, G or R, and X1, X3 and X5 are any amino acid, or
    • X2 is A, G or R, X5 is A, G or R, and X1, X3 and X4 are any amino acid, or
    • X3 is A, G or R, X4 is A, G or R, and X1, X2 and X5 are any amino acid, or
    • X3 is A, G or R, X5 is A, G or R, and X1, X2 and X4 are any amino acid, or
    • X4 is A, G or R, X5 is A, G or R, and X1, X2 and X3 are any amino acid,


      in particular:
    • X1 is R, X2 is R, and X3, X4 and X5 are any amino acid, or
    • X1 is R, X3 is R, and X2, X4 and X5 are any amino acid, or
    • X1 is R, X4 is R, and X2, X3 and X5 are any amino acid, or
    • X1 is R, X5 is R, and X2, X3 and X4 are any amino acid, or
    • X2 is R, X3 is R, and X1, X4 and X5 are any amino acid, or
    • X2 is R, X4 is R, and X1, X3 and X5 are any amino acid, or
    • X2 is R, X5 is R, and X1, X3 and X4 are any amino acid, or
    • X3 is R, X4 is R, and X1, X2 and X5 are any amino acid, or
    • X3 is R, X5 is R, and X1, X2 and X4 are any amino acid, or
    • X4 is R, X5 is R, and X1, X2 and X3 are any amino acid.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated immunosuppressive domain contains the amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 3, wherein: X1 is A, G or R, and X2, X3, X4 and X5 are any amino acid different from A, G or R, or

    • X2 is A, G or R, and X1, X3, X4 and X5 are any amino acid different from A, G or R, or
    • X3 is A, G or R, and X1, X2, X4 and X5 are any amino acid different from A, G or R, or
    • X4 is A, G or R, and X1, X2, X3 and X5 are any amino acid different from A, G or R, or
    • X5 is A, G or R, and X1, X2, X3 and X4 are any amino acid different from A, G or R,


      in particular:
    • X1 is R, and X2, X3, X4 and X5 are any amino acid different from A, G or R, or
    • X2 is R, and X1, X3, X4 and X5 are any amino acid different from A, G or R, or
    • X3 is R, and X1, X2, X4 and X5 are any amino acid different from A, G or R, or
    • X4 is R, and X1, X2, X3 and X5 are any amino acid different from A, G or R, or
    • X5 is R, and X1, X2, X3 and X4 are any amino acid different from A, G or R.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated immunosuppressive domain contains the amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 3, wherein:

    • X1 is A, G or R, X2 is A, G or R, and X3, X4 and X5 are any amino acid different from A, G or R, or
    • X1 is A, G or R, X3 is A, G or R, and X2, X4 and X5 are any amino acid different from A, G or R, or
    • X1 is A, G or R, X4 is A, G or R, and X2, X3 and X5 are any amino acid different from A, G or R, or
    • X1 is A, G or R, X5 is A, G or R, and X2, X3 and X4 are any amino acid different from A, G or R, or
    • X2 is A, G or R, X3 is A, G or R, and X1, X4 and X5 are any amino acid different from A, G or R, or
    • X2 is A, G or R, X4 is A, G or R, and X1, X3 and X5 are any amino acid different from A, G or R, or
    • X2 is A, G or R, X5 is A, G or R, and X1, X3 and X4 are any amino acid different from A, G or R, or
    • X3 is A, G or R, X4 is A, G or R, and X1, X2 and X5 are any amino acid different from A, G or R, or
    • X3 is A, G or R, X5 is A, G or R, and X1, X2 and X4 are any amino acid different from A, G or R, or
    • X4 is A, G or R, X5 is A, G or R, and X1, X2 and X3 are any amino acid different from A, G or R,


      in particular:
    • X1 is R, X2 is R, and X3, X4 and X5 are any amino acid different from A, G or R, or
    • X1 is R, X3 is R, and X2, X4 and X5 are any amino acid different from A, G or R, or
    • X1 is R, X4 is R, and X2, X3 and X5 are any amino acid different from A, G or R, or
    • X1 is R, X5 is R, and X2, X3 and X4 are any amino acid different from A, G or R, or
    • X2 is R, X3 is R, and X1, X4 and X5 are any amino acid different from A, G or R, or
    • X2 is R, X4 is R, and X1, X3 and X5 are any amino acid different from A, G or R, or
    • X2 is R, X5 is R, and X1, X3 and X4 are any amino acid different from A, G or R, or
    • X3 is R, X4 is R, and X1, X2 and X5 are any amino acid different from A, G or R, or
    • X3 is R, X5 is R, and X1, X2 and X4 are any amino acid, different from A, G or R or
    • X4 is R, X5 is R, and X1, X2 and X3 are any amino acid different from A, G or R.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X3 is R, X4 is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X4 is R, X3 is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X3 is R, X4 is R, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X3 is R, and X1 is E, X2 is K, X4 is L, X5 is Y, or
    • X4 is R, and X1 is E, X2 is K, X3 is F, X5 is Y, or
    • X3 is R, X4 is R, and X1 is E, X2 is K, X5 is Y.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X3 is R, and X1, X2, X4, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein:

    • X3 is R, and X1 is E, X2 is K, X4 is L, X5 is Y.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) contains the following amino acid sequence:









(SEQ ID NO: 489)









A-[I/M/T/L]-[A/G/R]-K-[F/P]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 490)









A-[I/M/T/L]-E-[A/G/R]-[F/P]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 491)









A-[I/M/T/L]-E-K-[A/G/R]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 492)









A-[I/M/T/L]-E-K-[F/P]-[A/G/R]-Y-T-A,



or










(SEQ ID NO: 493)









A-[I/M/T/L]-E-K-[F/P]-[L/V/I]-[A/G/R]-T-A,



or










(SEQ ID NO: 494)









A-[I/M/T/L]-E-K-[A/G/R]-[A/G/R]-Y-T-A,







in particular:









(SEQ ID NO: 489)









A-[I/M/T/L]-[A/G/R]-K-[F/P]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 495)









A-[I/M/T/L]-E-G-[F/P]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 491)









A-[I/M/T/L]-E-K-[A/G/R]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 492)









A-[I/M/T/L]-E-K-[F/P]-[A/G/R]-Y-T-A,



or










(SEQ ID NO: 496)









A-[I/M/T/L]-E-K-[F/P]-[L/V/I]-G-T-A,



or










(SEQ ID NO: 494)









A-[I/M/T/L]-E-K-[A/G/R]-[A/G/R]-Y-T-A.






In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) contains the following amino acid sequence:









(SEQ ID NO: 497)









A-[I/M/T/L]-R-K-[F/P]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 498)









A-[I/M/T/L]-E-R-[F/P]-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 499)









A-[I/M/T/L]-E-K-R-[L/V/I]-Y-T-A,



or










(SEQ ID NO: 500)









A-[I/M/T/L]-E-K-[F/P]-R-Y-T-A,



or










(SEQ ID NO: 501)









A-[I/M/T/L]-E-K-[F/P]-[L/V/I]-R-T-A,



or










(SEQ ID NO: 502)









A-[I/M/T/L]-E-K-R-R-Y-T-A.






In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said isolated mutated feline lentiviral ENV protein or said fragment thereof, comprises one of the amino acid sequences SEQ ID NO: 28 to 171.


In the invention “SEQ ID NO: 28 to 171” encompasses SEQ ID NO: 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170 and 171.


The correspondence is the following one:









SEQ ID NO: 28









AIEKRLYTA










SEQ ID NO: 29









AMEKRLYTA










SEQ ID NO: 30









ATEKRLYTA










SEQ ID NO: 31









ALEKRLYTA










SEQ ID NO: 32









AIEKRVYTA










SEQ ID NO: 33









AMEKRVYTA










SEQ ID NO: 34









ATEKRVYTA










SEQ ID NO: 35









ALEKRVYTA










SEQ ID NO: 36









AIEKRIYTA










SEQ ID NO: 37









AMEKRIYTA










SEQ ID NO: 38









ATEKRIYTA










SEQ ID NO: 39









ALEKRIYTA










SEQ ID NO: 40









AIEKGLYTA










SEQ ID NO: 41









AMEKGLYTA










SEQ ID NO: 42









ATEKGLYTA










SEQ ID NO: 43









ALEKGLYTA










SEQ ID NO: 44









AIEKGVYTA










SEQ ID NO: 45









AMEKGVYTA










SEQ ID NO: 46









ATEKGVYTA










SEQ ID NO: 47









ALEKGVYTA










SEQ ID NO: 48









AIEKGIYTA










SEQ ID NO: 49









AMEKGIYTA










SEQ ID NO: 50









ATEKGIYTA










SEQ ID NO: 51









ALEKGIYTA










SEQ ID NO: 52









AIEKLLYTA










SEQ ID NO: 53









AMEKLLYTA










SEQ ID NO: 54









ATEKLLYTA










SEQ ID NO: 55









ALEKLLYTA










SEQ ID NO: 56









AIEKLVYTA










SEQ ID NO: 57









AMEKLVYTA










SEQ ID NO: 58









ATEKLVYTA










SEQ ID NO: 59









ALEKLVYTA










SEQ ID NO: 60









AIEKLIYTA










SEQ ID NO: 61









AMEKLIYTA










SEQ ID NO: 62









ATEKLIYTA










SEQ ID NO: 63









ALEKLIYTA










SEQ ID NO: 64









AIEKALYTA










SEQ ID NO: 65









AMEKALYTA










SEQ ID NO: 66









ATEKALYTA










SEQ ID NO: 67









ALEKALYTA










SEQ ID NO: 68









AIEKAVYTA










SEQ ID NO: 69









AMEKAVYTA










SEQ ID NO: 70









ATEKAVYTA










SEQ ID NO: 71









ALEKAVYTA










SEQ ID NO: 72









AIEKLIYTA










SEQ ID NO: 73









AMEKAIYTA










SEQ ID NO: 74









ATEKAIYTA










SEQ ID NO: 75









ALEKAIYTA










SEQ ID NO: 76









AIEKFRYTA










SEQ ID NO: 77









AMEKFRYTA










SEQ ID NO: 78









ATEKFRYTA










SEQ ID NO: 79









ALEKFRYTA










SEQ ID NO: 80









AIEKPRYTA










SEQ ID NO: 81









AMEKPRYTA










SEQ ID NO: 82









ATEKPRYTA










SEQ ID NO: 83









ALEKPRYTA










SEQ ID NO: 84









AIEKFGYTA










SEQ ID NO: 85









AMEKFGYTA










SEQ ID NO: 86









ATEKFGYTA










SEQ ID NO: 87









ALEKFGYTA










SEQ ID NO: 88









AIEKPGYTA










SEQ ID NO: 89









AMEKPGYTA










SEQ ID NO: 90









ATEKPGYTA










SEQ ID NO: 91









ALEKPGYTA










SEQ ID NO: 92









AIEKFFYTA










SEQ ID NO: 93









AMEKFFYTA










SEQ ID NO: 94









ATEKFFYTA










SEQ ID NO: 95









ALEKFFYTA










SEQ ID NO: 96









AIEKPFYTA










SEQ ID NO: 97









AMEKPFYTA










SEQ ID NO: 98









ATEKPFYTA










SEQ ID NO: 99









ALEKPFYTA










SEQ ID NO: 100









AIEKFAYTA










SEQ ID NO: 101









AMEKFAYTA










SEQ ID NO: 102









ATEKFAYTA










SEQ ID NO: 103









ALEKFAYTA










SEQ ID NO: 104









AIEKPAYTA










SEQ ID NO: 105









AMEKPAYTA










SEQ ID NO: 106









ATEKPAYTA










SEQ ID NO: 107









ALEKPAYTA










SEQ ID NO: 108









AIEKRRYTA










SEQ ID NO: 109









AMEKRRYTA










SEQ ID NO: 110









ATEKRRYTA










SEQ ID NO: 111









ALEKRRYTA










SEQ ID NO: 112









AIEKRAYTA










SEQ ID NO: 113









AMEKRAYTA










SEQ ID NO: 114









ATEKRAYTA










SEQ ID NO: 115









ALEKRAYTA










SEQ ID NO: 116









AIEKRFYTA










SEQ ID NO: 117









AMEKRFYTA










SEQ ID NO: 118









ATEKRFYTA










SEQ ID NO: 119









ALEKRFYTA










SEQ ID NO: 120









AIEKRGYTA










SEQ ID NO: 121









AMEKRGYTA










SEQ ID NO: 122









ATEKRGYTA










SEQ ID NO: 123









ALEKRGYTA










SEQ ID NO: 124









AIEKARYTA










SEQ ID NO: 125









AMEKARYTA










SEQ ID NO: 126









ATEKARYTA










SEQ ID NO: 127









ALEKARYTA










SEQ ID NO: 128









AIEKAAYTA










SEQ ID NO: 129









AMEKAAYTA










SEQ ID NO: 130









ATEKAAYTA










SEQ ID NO: 131









ALEKAAYTA










SEQ ID NO: 132









AIEKAFYTA










SEQ ID NO: 133









AMEKAFYTA










SEQ ID NO: 134









ATEKAFYTA










SEQ ID NO: 135









ALEKAFYTA










SEQ ID NO: 136









AIEKAGYTA










SEQ ID NO: 137









AMEKAGYTA










SEQ ID NO: 138









ATEKAGYTA










SEQ ID NO: 139









ALEKAGYTA










SEQ ID NO: 140









AIEKGRYTA










SEQ ID NO: 141









AMEKGRYTA










SEQ ID NO: 142









ATEKGRYTA










SEQ ID NO: 143









ALEKGRYTA










SEQ ID NO: 144









AIEKGAYTA










SEQ ID NO: 145









AMEKGAYTA










SEQ ID NO: 146









ATEKGAYTA










SEQ ID NO: 147









ALEKGAYTA










SEQ ID NO: 148









AIEKGFYTA










SEQ ID NO: 149









AMEKGFYTA










SEQ ID NO: 150









ATEKGFYTA










SEQ ID NO: 151









ALEKGFYTA










SEQ ID NO: 152









AIEKGGYTA










SEQ ID NO: 153









AMEKGGYTA










SEQ ID NO: 154









ATEKGGYTA










SEQ ID NO: 155









ALEKGGYTA










SEQ ID NO: 156









AIEKFRYTA










SEQ ID NO: 157









AMEKFRYTA










SEQ ID NO: 158









ATEKFRYTA










SEQ ID NO: 159









ALEKFRYTA










SEQ ID NO: 160









AIEKFAYTA










SEQ ID NO: 161









AMEKFAYTA










SEQ ID NO: 162









ATEKFAYTA










SEQ ID NO: 163









ALEKFAYTA










SEQ ID NO: 164









AIEKFFYTA










SEQ ID NO: 165









AMEKFFYTA










SEQ ID NO: 166









ATEKFFYTA










SEQ ID NO: 167









ALEKFFYTA










SEQ ID NO: 168









AIEKFGYTA










SEQ ID NO: 169









AMEKFGYTA










SEQ ID NO: 170









ATEKFGYTA










SEQ ID NO: 171









ALEKFGYTA






In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said fragment of said isolated mutated feline lentiviral ENV protein comprises at least 40 amino acids, in particular at least 60 amino acids.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said isolated mutated feline lentiviral ENV protein or said fragment thereof, comprises one of the amino acid sequences SEQ ID NO: 172 to 315.


In the invention “SEQ ID NO: 172 to 315” encompasses SEQ ID NO: 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314 and 315.


The correspondence is the following one:









SEQ ID NO: 172









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKRLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 173









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKRLY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 174









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKRLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 175









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKRLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 176









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKRVYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 177









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKRVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 178









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKRVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 179









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKRVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 180









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKRIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 181









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKRIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 182









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKRIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 183









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKRIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 184









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKGLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 185









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKGLY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 186









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKGLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 187









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKGLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 188









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKGVYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 189









AAIEKVTGALKINNLRLVTLEHQVLVIGLKVEMEKGVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 190









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKGVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 191









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKGVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 192









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKGIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 193









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKGIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 194









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKGIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 195









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKGIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 196









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKLLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 197









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKLLY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 198









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKLLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 199









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKLLYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 200









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKLVYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 201









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKLVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 202









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKLVYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 203









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKLVYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 204









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKLIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 205









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKLIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 206









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKLIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 207









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKLIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 208









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKALYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 209









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKALY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 210









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKALYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 211









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKALYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 212









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKAVYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 213









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKAVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 214









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKAVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 215









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKAVY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 216









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKAIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 217









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKAIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 218









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKAIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 219









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKAIYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 220









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 221









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFRY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 222









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 223









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 224









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKPRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 225









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKPRY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 226









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKPRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 227









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKPRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 228









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 229









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 230









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 231









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 232









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKPGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 233









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKPGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 234









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKPGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 235









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKPGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 236









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 237









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFFY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 238









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 239









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 240









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKPFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 241









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKPFY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 242









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKPFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 243









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKPFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 244









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 245









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 246









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 247









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 248









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKPAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 249









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKPAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 250









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKPAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 251









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKPAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 252









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKRRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 253









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKRRY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 254









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKRRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 255









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKRRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 256









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKRAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 257









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKRAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 258









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKRAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 259









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKRAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 260









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKRFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 261









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKRFY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 262









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKRFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 263









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKRFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 264









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKRGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 265









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKRGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 266









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKRGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 267









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKRGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 268









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKARYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 269









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKARY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 270









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKARY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 271









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKARY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 272









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKAAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 273









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKAAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 274









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKAAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 275









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKAAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 276









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKAFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 277









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKAFY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 278









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKAFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 279









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKAFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 280









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKAGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 281









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKAGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 282









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKAGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 283









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKAGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 284









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKGRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 285









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKGRY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 286









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKGRY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 287









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKGRY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 288









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKGAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 289









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKGAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 290









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKGAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 291









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKGAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 292









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKGFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 293









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKGFY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 294









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKGFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 295









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKGFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 296









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKGGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 297









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKGGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 298









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKGGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 299









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKGGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 300









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 301









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFRY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 302









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 303









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFRYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 304









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 305









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFAY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 306









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 307









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFAYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 308









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 309









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFFY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 310









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 311









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFFYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 312









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAIEKFGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 313









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFGY






TAFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 314









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEATEKFGYT






AFAMQELGCNQNQFFCKIPLELWTR










SEQ ID NO: 315









AIEKVTGALKINNLRLVTLEHQVLVIGLKVEALEKFGYT






AFAMQELGCNQNQFFCKIPLELWTR






As mentioned above, the previous ENV proteins having their ISU comprising the above sequence have decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties.


Thus, in other words, any feline lentiviral Env protein having in their ISU an amino acid sequence comprising the sequences SEQ ID NO: 28 to 315, have decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties.


In other words, a feline lentiviral ENV protein comprising, within its ISU domain, an amino acid sequence selected from SEQ ID NO: 28 to 315 have decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties.


In the invention, the fragments of the mutated ENV proteins according to the invention have decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties.


However, these fragments retain the structure and the antigenicity of the corresponding immunosuppressive domain that is not mutated, i.e. the wild type immunosuppressive domain.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein mutated feline lentiviral ENV protein or fragment thereof further comprises an additional mutation of one at least of the amino acids X1*, X2*,


X3*, X4* and X5* in the following sequence:









(SEQ ID NO: 316)









X1*-X2*-X3*-X4*-X5*-[V/I]-[E/R]-A-[I/M/T/L]-X1-






X2-X3-X4-X5-T-A







wherein
    • X1, X2, X3, X4, X5 are defined as above, and
    • X1* is either deleted, or is A, C, D, E, F, G, H, I, K, L, N, P, Q, R, S, Y or W, in particular A, F, G, L or R, and/or
    • X2* is either deleted, or is A, C, D, E, F, G, H, K, M, N, P, Q, R, S, T, V, Y or W, in particular A, F, G, or R, and/or
    • X3* is either deleted, or is A, C, D, E, F, L, H, I, K, M, N, P, Q, R, S, T, V,


Y or W, in particular A, F, L or R, and/or

    • X4* is either deleted, or is A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, Y or W, in particular A, F, G, or R, and/or
    • X5* is either deleted, or is A, C, D, E, F, G, L, H, I, M, N, P, Q, S, V, Y or W, in particular A, F, G or L,


      in association with a pharmaceutically acceptable carrier.


In such a pharmaceutical composition, the mutated feline lentiviral ENV protein, or fragment thereof, comprises a mutation of one at least of the amino acids X1, X2, X3, X4 and X5 in association with an additional mutation of one at least of the amino acids X1*, X2*, X3*, X4*and X5*.


In the invention, “a mutation of one at least of the amino acids X1*, X2*, X3*, X4* and X5*” means that said mutated acids are:

    • X1* or X2* or X3* or X4* or X5* or
    • X1*/X2* or X1*/X3* or X1*/X4* or X1*/X5* or X2*/X3* or X2*/X4* or X2*/X5* or X3*/X4* or X3*/X5* or X4*/X5* or
    • X1*/X2*/X3* or X1*/X2*/X4* or X1*/X2*/X5* or X1*/X3*/X4* or X1*/X3*/X5* or
    • X1*/X4*/X5* or X2*/X3*/X4* or X2*/X3*/X5* or X2*/X4*/X5* or X3*/X4*/X5* or
    • X1*/X2*/X3*/X4* or X1*/X2*/X3*/X5* or X1*/X2*/X4*/X5* or X1*/X3*/X4*/X5* or X2*/X3*/X4*/X5* or
    • X1*/X2*/X3*/X4*/X5*.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein mutated feline lentiviral ENV protein or fragment thereof further comprises an additional mutation of the amino acids X3* in the following sequence:









(SEQ ID NO: 434)









[T/V/M]-[L/I]-X3*-L-[K/T/R]-[V/I]-[E/R]-A-






[I/M/T/L]-X1-X2-X3-X4-X5-T-A







wherein
    • X1, X2, X3, X4, X5 are defined as above, and
    • X3* is either deleted, or is A, C, D, E, F, L, H, I, K, M, N, P, Q, R, S, T, V, Y or W, in particular A, F, L or R,


      in association with a pharmaceutically acceptable carrier.


In such a pharmaceutical composition, the mutated feline lentiviral ENV protein, or fragment thereof, comprises a mutation of one at least of the amino acids X1, X2, X3, X4 and X5 in association with an additional mutation of X3*.


The sequences SEQ ID NO: 316 and SEQ ID NO: 434 contain the following amino acid sequence A[I/M/T/L]X1X2X3X4X5TA (SEQ ID NO: 1) elongated on its N-terminal end, in which an additional mutation is present.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance an isolated mutated feline lentiviral ENV protein having substantially no immunosuppressive activity, or a fragment thereof,


said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,


said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 467)









X1*X2*-X3*-X4*-X5*-[V/I]-[E/R]-A-[I/M/T/L]-E-K-






[F/P]-[L/V/I]-Y-T-A 







wherein
    • X1* is either deleted, or is A, C, D, E, F, G, H, I, K, L, N, P, Q, R, S, T V Y or W, in particular A, F, G, L or R, and/or
    • X2* is either deleted, or is A, C, D, E, F, G, H, K, M, N, P, Q, R, S, T, V, Y or W, in particular A, F, G or R, and/or
    • X3* is either deleted, or is A, C, D, E, F, L, H, I, K, M, N, P, Q, R, S, T, V, Y or W, in particular A, F, L or R, and/or
    • X4* is either deleted, or is A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, Y or W, in particular A, F, G, or R, and/or
    • X5* is either deleted, or is A, C, D, E, F, G, L, H, I, M, N, P, Q, S, V, Y or W, in particular A, F, G, or L.


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance an isolated mutated feline lentiviral ENV protein having substantially no immunosuppressive activity, or a fragment thereof,


said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,


said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 468)









[T/V/M]-[L/I]-X3*-L-[K/T/R]-[V/I]-[E/R]-A-






[I/M/T/L]-E-K-[F/P]-[L/V/I]-Y-T-A







wherein
    • X3* is either deleted, or is A, C, D, E, F, L, H, I, K, M, N, P, Q, R, S, T, V, Y or W, in particular A, F, L or R,


      in association with a pharmaceutically acceptable carrier.


In another advantageous embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated feline lentiviral protein, said variant or said fragment harbour a three-dimensional structure similar to the structure of the natural non mutated feline lentiviral ENV protein, non mutated variant or non fragment thereof.


The skilled person knows how to measure the antigenicity, by using standard proceedings.


In another advantageous embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated feline lentiviral protein or said fragment thereof are expressed at the plasma membrane at a level substantially identical to the expression at the plasma membrane of the natural non mutated lentiviral ENV protein or non mutated fragment thereof.


The membrane expression of the lentiviral ENV protein according to the invention can be measured by any techniques allowing determination of a plasma membrane protein. For instance, cells can be transfected with an expression vector allowing the expression of the mutated lentiviral ENV protein according to the invention. Cells are then incubated with an antibody recognizing specifically the extracellular part of said lentiviral mutated ENV protein. The complex (antibody/ENV protein) is detected by another antibody, and the complex can be quantified by flow cytometry (see examples).


If no complex is detected, the mutated ENV protein is not expressed at the plasma membrane. On the contrary, if the complex is detected, this means that the mutated ENV protein is expressed at the plasma membrane and in an appropriate conformation so as to be detected by the antibody recognizing the extracellular part of the protein.


In another advantageous embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated feline lentiviral ENV protein is such that it has conserved, totally or partially, its fusogenic activity, which is responsible for virus-cell or cell-cell membrane fusion and can be measured by appropriate assays.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said isolated mutated feline lentiviral ENV protein consists of one of the amino acid sequences: SEQ ID NO: 5 and SEQ ID NO: 317 to 342 and 374 to 419.









SEQ ID NO: 317









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKGLYTAFAMQELGCNQNQFFC






KIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQK






FYEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYL






KGLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIA






MPEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 318









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKLLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 319









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAPP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKALYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 320









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFRYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 321









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFGYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 322









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFAYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 323









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFFYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 324









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMRKFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 325









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMGKFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 326









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMLKFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 327









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMAKFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 328









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMFKFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 329









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMERFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 330









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEGFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 331









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMELFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 332









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEAFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 333









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEFFLYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 334









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFLRTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 335









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFLGTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 336









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFLLTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 337









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFLATAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 338









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKFLFTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 339









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKRRYTAFAMQELGCNQNQFFC






KIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQK






FYEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYL






KGLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIA






MPEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 340









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKRGYTAFAMQELGCNQNQFFC






KIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQK






FYEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYL






KGLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIA






MPEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 341









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKRAYTAFAMQELGCNQNQFFC






KIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQK






FYEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYL






KGLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIA






MPEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 342









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDERVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFTGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






EEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQ






RNRWEWRPDFKSKKVKISLPCNSTKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSNLHTEARFRIRCRWNVGSDTSLIDT






CGNTPNVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDL






IVHFNMTKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTN






SSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVV






KQPDYLLVPEEVMEYKPRRKRAAIHVMLALATVLSIAGA






GTGATAIGMVTQYHQVLATHQEAIEKVTGALKINNLRLV






TLEHQVLVIGLKVEAMEKRFYTAFAMQELGCNQNQFFCK






IPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKF






YEIIMDIEQNNVQGKTGIQQLQKWEDWVRWIGNIPQYLK






GLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAM






PEVEGEEIQPQMELRRNGRQCGMSEKEE










SEQ ID NO: 374









MAEGFAANRQWIGLEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDESVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFIGIIIYSQTTNAQVVWRLPPLVVPVEESEIIFWDCWAPEE






PACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATLFK






KATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIVPD






YQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLSYC






TDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQM






AYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQRN






RWEWRPDFESKKVKISLQCNSTKNLTFAMRSSGDYGEVT






GAWIEFGCHRNKSKLHAEARFRIRCRWNVGSNTSLIDTCG






NTQKVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDLIM






HFNMKKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTNSS






SSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVVKQ






PDYLVVPEEVMEYKPRRKRAAIHVMLALAAVLSIAGAGT






GATAIGMVTQYHQVLATHQEAVEKVTEALKINNLRLVTL






EHQVLVIGLKVEAMEKRLYTAFAMQELGCNQNQFFCKIP






PELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYE






IIMDIEQNNVQGKKGIQQLQKWEDWVGWIGNIPQYLKGL






LGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAMPE






VEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 375









MAEGFAANRQWIGLEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRNEIQEVKLEEGN






AGKFRRARFLRYSDESVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEECYNNREKGTTDNIKYGRRCCLGTVTLYLI






LFIGIIIYSQTTNAQVVWRLPPLVVPVEESEIIFWDCWAPEE






PACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATLFK






KATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIVPD






YQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLSYC






TDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQM






AYYNSCKWEEAKVKFHCQRTQSQPGSWFRAISSWKQRN






RWEWRPDFESKKVKISLQCNSTKNLTFAMRSSGDYGEVT






GAWIEFGCHRNKSKLHAEARFRIRCRWNVGSNTSLIDTCG






NTQKVSGANPVDCTMYSNKMYNCSLQNGFTMKVDDLIM






HFNMKKAVEMYNIAGNWSCTSDLPSSWGYMNCNCTNSS






SSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQVVKQ






PDYLVVPEEVMEYKPRRKRAAIHVMLALAAVLSIAGAGT






GATAIGMVTQYHQVLATHQEAVEKVTEALKINNLRLVTL






EHQVLVIGLKVEAMEKFRYTAFAMQELGCNQNQFFCKIP






PELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYE






IIMDIEQNNVQGKKGIQQLQKWEDWVGWIGNIPQYLKGL






LGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAMPE






VEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 376









MAEGFVANGQWIGPEEAEELVDFEIATQMNEEGPLNPGIN






PFRVPGITKQEKQEYCSTMQPKLQALRNEIQEVKLEEGNA






GKFRRARFLRYSDETILSLIYLFIGYFRYLVDRKRFGSLRH






DIDIEAPQEECYNNKEKGMTENIKYGKRCLVGTAALYLIL






AIGIIIIIRTTDAQVVWRLPPLVVPVEESEIIFWDCWAPEEPA






CQDFLGAMIHLKASTNISNTEGPTLGNWAREIWATLFKKA






TRQCRRGRIWKRWNETITGPIGCANNTCYNISVIVPDYQC






YIDRVDTWLQGKVNISLCLTGGKMLYNKETKQLSYCTDP






LQIPLINYTFGPNQTCMWNISQIQDPEIPKCGWWNQQAYY






NNCKWERTDVKFQCQRTQSQPGSWIRAISSWKQGNRWE






WRPDFESERVKVSLQCNSTRNLTFAMRSSGDYGEITGAWI






EFGCHRNKSIRHNAARFRIRCRWNEGDNNSLIDTCGETQN






VSGANPVDCTMYANKMYNCSLQDGFTMKVDDLIMHFN






MTKAVEMYNIAGNWSCMSDLPTEWGYMNCNCTNDTSN






NNTRKMKCPKENGILRNWYNPVAGLRQSLEKYQVVKQP






DYLLVPEEVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATQQEAIEKVTEALKITNLRLVTLE






HQVLVIGLKVEAMEKRLYTAFAMQELGCNQNQFFCKVPP






ELWRRYNMTINQTIWNHGNITLGEWYNQTKDLQKKFYGI






IMDIEQNNVQGKKGLQQLQKWEDWVGWIGNIPQYLKGL






LGSIVGIGLGILLLILCLPTLVDCIRNCIHKILGYTVIAMPEV






DGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 377









MAEGFVANGQWIGPEEAEELVDFEIATQMNEEGPLNPGIN






PFRVPGITKQEKQEYCSTMQPKLQALRNEIQEVKLEEGNA






GKFRRARFLRYSDETILSLIYLFIGYFRYLVDRKRFGSLRH






DIDIEAPQEECYNNKEKGMTENIKYGKRCLVGTAALYLIL






AIGIIIIIRTTDAQVVWRLPPLVVPVEESEIIFWDCWAPEEPA






CQDFLGAMIHLKASTNISNTEGPTLGNWAREIWATLFKKA






TRQCRRGRIWKRWNETITGPIGCANNTCYNISVIVPDYQC






YIDRVDTWLQGKVNISLCLTGGKMLYNKETKQLSYCTDP






LQIPLINYTFGPNQTCMWNISQIQDPEIPKCGWWNQQAYY






NNCKWERTDVKFQCQRTQSQPGSWIRAISSWKQGNRWE






WRPDFESERVKVSLQCNSTRNLTFAMRSSGDYGEITGAWI






EFGCHRNKSIRHNAARFRIRCRWNEGDNNSLIDTCGETQN






VSGANPVDCTMYANKMYNCSLQDGFTMKVDDLIMHFN






MTKAVEMYNIAGNWSCMSDLPTEWGYMNCNCTNDTSN






NNTRKMKCPKENGILRNWYNPVAGLRQSLEKYQVVKQP






DYLLVPEEVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATQQEAIEKVTEALKITNLRLVTLE






HQVLVIGLKVEAMEKFRYTAFAMQELGCNQNQFFCKVPP






ELWRRYNMTINQTIWNHGNITLGEWYNQTKDLQKKFYGI






IMDIEQNNVQGKKGLQQLQKWEDWVGWIGNIPQYLKGL






LGSIVGIGLGILLLILCLPTLVDCIRNCIHKILGYTVIAMPEV






DGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 378









MAEGFVANGQWIGPEEAEELLDFDIATQLNEEGPLNPGIN






PFRVPGITQKEKQEYCNTLQPKLQALRNEIQEVKLEERNA






GKFRRARFLRYSDETILSLIYSFVGYFRYLVDRKKFGSLRH






DIDIEAPQEEYYNNKEKGMTDNVKYGKRCLVGTAAFYLL






LAIGIIIIIRTVDAQVVWRLPPLVVPVEESEIIFWDCWAPEEP






ACQDFLGAMIHLKASTNIRIQEGPTLGNWAREIWATLFKK






ATRQCRRGRIWKRWNETITGPIGCANNTCYNISVIIPDYQC






YIDRVDTWLQGKVNISICLTGGKMLYNKETKQLSYCTDP






LQIPLINYTFGPQQTCMWNTSQIQDPEIPKCGWWNQKAY






YNQCSWEQTDVKFQCQRTQSQPGSWIRAISSWRQRNRWE






WRPDFESERVKVSLQCNSTQNLTFAMRSSGDYGEITGAWI






EFGCHRNKSKHHNEARFRIRCRWNEGNNNSLIDTCGKTQ






NVLGANPVDCTMYANRMYNCSLQDGFTMKVDDLIMHF






NMTKAVEMYNIAGNWSCMSDLPTNWGYMKCNCTNDTS






NNHTIKMECPEEKGILRNWYNPVAGLRQSLEKYQVVKQP






DYLVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATHQEAIEKVTEALKINNLRLVTLE






HQVLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKVPL






ELWRRYNMTINQTIWNHGNITLGEWYNQTKALQHKFYEI






IMDIEQNNVQGKKGLQQLQKWEDWVGWIGNIPQYLKGL






LGGILGIGLGILLLILCLPTLVDCIRNCIHKILGYTVIAMPDV






DEEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 379









MAEGFVANGQWIGPEEAEELLDFDIATQLNEEGPLNPGIN






PFRVPGITQKEKQEYCNTLQPKLQALRNEIQEVKLEERNA






GKFRRARFLRYSDETILSLIYSFVGYFRYLVDRKKFGSLRH






DIDIEAPQEEYYNNKEKGMTDNVKYGKRCLVGTAAFYLL






LAIGIIIIIRTVDAQVVWRLPPLVVPVEESEIIFWDCWAPEEP






ACQDFLGAMIHLKASTNIRIQEGPTLGNWAREIWATLFKK






ATRQCRRGRIWKRWNETITGPIGCANNTCYNISVIIPDYQC






YIDRVDTWLQGKVNISICLTGGKMLYNKETKQLSYCTDP






LQIPLINYTFGPQQTCMWNTSQIQDPEIPKCGWWNQKAY






YNQCSWEQTDVKFQCQRTQSQPGSWIRAISSWRQRNRWE






WRPDFESERVKVSLQCNSTQNLTFAMRSSGDYGEITGAWI






EFGCHRNKSKHHNEARFRIRCRWNEGNNNSLIDTCGKTQ






NVLGANPVDCTMYANRMYNCSLQDGFTMKVDDLIMHF






NMTKAVEMYNIAGNWSCMSDLPTNWGYMKCNCTNDTS






NNHTIKMECPEEKGILRNWYNPVAGLRQSLEKYQVVKQP






DYLVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATHQEAIEKVTEALKINNLRLVTLE






HQVLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKVPL






ELWRRYNMTINQTIWNHGNITLGEWYNQTKALQHKFYEI






IMDIEQNNVQGKKGLQQLQKWEDWVGWIGNIPQYLKGL






LGGILGIGLGILLLILCLPTLVDCIRNCIHKILGYTVIAMPDV






DEEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 380









MAEGFAANRQWIGPEEAEELLDFDIAIQMNEEGPLNPGVN






PFRVPGITEAEKQEYCNILQPKLQDLKGKIQEVKLEEGNA






GKFRRARFLRYSDETVLSLIHLFIGYCPHLCRRHELGSLRH






DIDIEALQEERYNDREKGITDNIKYGKRCLIGTAVLYLLLS






LGIIIHTCKAQVVWRLPPLVVPVEESEIIFWDCWAPEEPAC






QDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKAT






RQCRRGRIWRRWNETITGPLGCANNTCYNISVIVPDYQCY






LDRVDTWLQGKVNISLCLTGGKMLYNKETKQLSYCTDPL






QIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQNAYY






NSCRWEHTDVQFQCQRTQSQPGSWIRAISSWKQRNRWE






WRPDFESEKVKVSLQCNSTKNLTFAMRSSGDYGEVTGA






WIEFGCHRTKSKYHTEARFRIRCRWNVGDNTSLIDTCGET






QNVSRANPVDCTMYANRMYNCSLQNGFTMKVDDLIMHF






NKTKAVEMYNIAGNWSCKSDLPPTWGYMNCNCTNSTNS






GTGIRMACPRNQGILRNWYNPVAGLRQSLEKYQVVKQPD






YLVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGTG






ATAIGMVTQYQQVLATHQEAIEKVTEALKINNLRLVTLEH






QVLVIGLKVEAMEKRLYTAFAMQELGCNQNQFFCKVPSA






LWERYNMTINQTIWNHGNITLGEWYNQTKDLQQRFYEII






MDIEQNNVQGKKGLQQLQEWEDWVGWIGNIPQYLKGLL






GGILGIGLGMLLLILCLPTLVDCIRNCIHKILGYTVIAMPEV






EEEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 381









MAEGFAANRQWIGPEEAEELLDFDIAIQMNEEGPLNPGVN






PFRVPGITEAEKQEYCNILQPKLQDLKGKIQEVKLEEGNA






GKFRRARFLRYSDETVLSLIHLFIGYCPHLCRRHELGSLRH






DIDIEALQEERYNDREKGITDNIKYGKRCLIGTAVLYLLLS






LGIIIHTCKAQVVWRLPPLVVPVEESEIIFWDCWAPEEPAC






QDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKAT






RQCRRGRIWRRWNETITGPLGCANNTCYNISVIVPDYQCY






LDRVDTWLQGKVNISLCLTGGKMLYNKETKQLSYCTDPL






QIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQNAYY






NSCRWEHTDVQFQCQRTQSQPGSWIRAISSWKQRNRWE






WRPDFESEKVKVSLQCNSTKNLTFAMRSSGDYGEVTGA






WIEFGCHRTKSKYHTEARFRIRCRWNVGDNTSLIDTCGET






QNVSRANPVDCTMYANRMYNCSLQNGFTMKVDDLIMHF






NKTKAVEMYNIAGNWSCKSDLPPTWGYMNCNCTNSTNS






GTGIRMACPRNQGILRNWYNPVAGLRQSLEKYQVVKQPD






YLVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGTG






ATAIGMVTQYQQVLATHQEAIEKVTEALKINNLRLVTLEH






QVLVIGLKVEAMEKFRYTAFAMQELGCNQNQFFCKVPSA






LWERYNMTINQTIWNHGNITLGEWYNQTKDLQQRFYEII






MDIEQNNVQGKKGLQQLQEWEDWVGWIGNIPQYLKGLL






GGILGIGLGMLLLILCLPTLVDCIRNCIHKILGYTVIAMPEV






EEEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 382









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRDEIQEVKLEEGN






AGKFRRTRFLRYSDEHVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEEYYNNREKGTTDNIKYGRRCCLGTVTLYL






ILFIGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






DEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNRCKWEEAKVKFHCQRTQSQPGSWRRAISSWKQ






RNRWEWRPDLESEKVKISLQCNSKKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSKHHSEARFRIRCRWNVGSNTSLIDT






CGNTQDVSGANPVDCTMYSNKMYNCSLQNGFTMKVDD






LIVHFSMTKAVKMYNIAGNWSCTSDLPSSWGYMNCNCT






NSSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQV






VKQPDYLVVPEEVMEYKPRRKRAAIHVMLALATVLSIAG






AGTGATAIGMVTQYHQVLATHQEAIEKVTEALKINNLRL






VTLEHQVLVIGLKVEAMEKRLYTAFAMQELGCNQNQFFC






KIPPGLWTRYNMTINQTIWNHGNITLGEWYNKTKDLQQK






FYEIIMDIEQNNVQGKTGIQQLQKWEDWVGWIGNIPQYL






KGLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIA






MPEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 383









MAEGFAANRQWIGPEEAEELLDFDIATQMSEEGPLNPGV






NPFRVPGITEKEKQNYCNILQPKLQDLRDEIQEVKLEEGN






AGKFRRTRFLRYSDEHVLSLVHAFIGYCIYLGNRNKLGSL






RHDIDIEAPQEEYYNNREKGTTDNIKYGRRCCLGTVTLYL






ILFIGVIVYSQTAGAQVVWRLPPLVVPVEESEIIFWDCWAP






DEPACQDFLGAMIHLKAKTNISIREGPTLGNWAREIWATL






FKKATRQCRRGRIWKRWNETITGPSGCANNTCYNVSVIV






PDYQCYLDRVDTWLQGKINISLCLTGGKMLYNKVTKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWN






QMAYYNRCKWEEAKVKFHCQRTQSQPGSWRRAISSWKQ






RNRWEWRPDLESEKVKISLQCNSKKNLTFAMRSSGDYGE






VTGAWIEFGCHRNKSKHHSEARFRIRCRWNVGSNTSLIDT






CGNTQDVSGANPVDCTMYSNKMYNCSLQNGFTMKVDD






LIVHFSMTKAVKMYNIAGNWSCTSDLPSSWGYMNCNCT






NSSSSYSGTKMACPSNRGILRNWYNPVAGLRQSLEQYQV






VKQPDYLVVPEEVMEYKPRRKRAAIHVMLALATVLSIAG






AGTGATAIGMVTQYHQVLATHQEAIEKVTEALKINNLRL






VTLEHQVLVIGLKVEAMEKFRYTAFAMQELGCNQNQFFC






KIPPGLWTRYNMTINQTIWNHGNITLGEWYNKTKDLQQK






FYEIIMDIEQNNVQGKTGIQQLQKWEDWVGWIGNIPQYL






KGLLGGILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIA






MPEVEGEEIQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 384









MAKGFAANRQWIGPEEAEELLDFDIATQMNEEGPLNPGV






NPFRVPGITEQEKQDYCNILQPKLQELRNEIQEVKLEEGNA






GKFRRARFLRYSDETILSLIHLFIGYCTYLCKRNELGSLRH






DIDIEAPQEECYNNKEKGTTNNIKYGGRCFIGTMIMYLLIFI






GIIIYIQTTEAQVVWRLPPLIVPVKESEIIFWDCWAPEEPAC






QDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKAT






RQCRRGRIWKRWNETITGPLGCANNTCYNISVIVPDYQCY






LDRVDTWLQGKVNISLCLTGGKMLYNKDTKQLSYCTDPL






QIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQIAYYN






SCKWEKTDVKFHCQRTQSQPGSWIRAISSWRQRNRWEW






RPDFESEKVKVSLQCNSTKNLTFAMRSSGDYGEVTGAWI






EFGCHRKKSKLHTEARFRIRCRWNVGNNASLIDTCGNTPD






VSGANPVNCTMYANKMYNCWLQNGFTIKVDDLIMHFN






MTKAVEMYNIAGNWSCTSDLPPTWGYMKCNCTNNSDDT






RGKMACPRTQGILRNWYNPVAGLRQSLEKYQVVKQPDY






LVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGTGA






TAIGMVTQYHQVLATHQEAIEKVTEALKINNLRLVTLEHQ






VLVIGLKVEAMEKRLYTAFAMQELGCNQNQFFCKVPPQL






WKRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEIIM






DIEQNNVQGKTGIQQLQRWEDWVGWIGNIPQYLKGLLGG






ILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAMPEVEEE






EIQQQMELRRNGRQCGMSEKEEE










SEQ ID NO: 385









MAKGFAANRQWIGPEEAEELLDFDIATQMNEEGPLNPGV






NPFRVPGITEQEKQDYCNILQPKLQELRNEIQEVKLEEGNA






GKFRRARFLRYSDETILSLIHLFIGYCTYLCKRNELGSLRH






DIDIEAPQEECYNNKEKGTTNNIKYGGRCFIGTMIMYLLIFI






GIIIYIQTTEAQVVWRLPPLIVPVKESEIIFWDCWAPEEPAC






QDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKAT






RQCRRGRIWKRWNETITGPLGCANNTCYNISVIVPDYQCY






LDRVDTWLQGKVNISLCLTGGKMLYNKDTKQLSYCTDPL






QIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQIAYYN






SCKWEKTDVKFHCQRTQSQPGSWIRAISSWRQRNRWEW






RPDFESEKVKVSLQCNSTKNLTFAMRSSGDYGEVTGAWI






EFGCHRKKSKLHTEARFRIRCRWNVGNNASLIDTCGNTPD






VSGANPVNCTMYANKMYNCWLQNGFTIKVDDLIMHFN






MTKAVEMYNIAGNWSCTSDLPPTWGYMKCNCTNNSDDT






RGKMACPRTQGILRNWYNPVAGLRQSLEKYQVVKQPDY






LVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGTGA






TAIGMVTQYHQVLATHQEAIEKVTEALKINNLRLVTLEHQ






VLVIGLKVEAMEKFRYTAFAMQELGCNQNQFFCKVPPQL






WKRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEIIM






DIEQNNVQGKTGIQQLQRWEDWVGWIGNIPQYLKGLLGG






ILGIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAMPEVEEE






EIQQQMELRRNGRQCGMSEKEEE










SEQ ID NO: 386









MAEGFAANRQWIGPEEAEELLDFDIATQMNEEGPLNPGIN






PFRVPGITEIEKQAYCKILQPRLQALRNEIQEVKLEEGNAG






KFRRARFLRYSDVAILSLIHVFIGYCTYLCNQQKLGSLRHD






IDIEAPQEEYYSNNEKGTTDNIKYGRRCVIGTVALYLLICT






GIIIYTRTATAQVVWRLPPLVVPVEESEVIFWDCWAPEEPA






CQDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKA






TRQCRRGRIWRRWNETITGPLGCANNTCYNISVIVPDYQC






YLDRVDTWLQGKINISLCLTGGKMLYNRYTKQLSYCTDP






LQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQRAY






YNSCRWESTDVKFHCQRTQSQPGSWLRAISSWRQRNRW






EWRPDFESEKVKVSLQCNSTKNLTFAMRSSGDYGDVTGA






WIEFGCHRNKSRLHTEARFRIRCRWNVGDNTSLIDTCGKT






QNIAGANPVDCTMYVNRMYNCSLQNGFTMKVDDLIMHF






NMTKAVEMYNIAGNWSCTSNLPPTWGYINCNCTNSSDSN






KMACPSSQGILRNWYNPVAGLRQSLEKYQVVKQPDYLV






VPGEVMEYKPRRKRAAIHVMLALATILSMAGAGTGATAI






GMVTQYHQVLATHQEAIEKVTEALKVNNLRLITLEHQVL






VIGLKVEAMEKRLYTAFAMQELGCNQNQFFCKVPPELW






KRYNMTINQTIWNHGNITLGEWYNQTKGLQQKFYEIIMDI






EQNSVQGKKGIQQLQEWEDWIGWIGNIPQYLKGLLGGIL






GIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAMPEVEGEE






IQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 387









MAEGFAANRQWIGPEEAEELLDFDIATQMNEEGPLNPGIN






PFRVPGITEIEKQAYCKILQPRLQALRNEIQEVKLEEGNAG






KFRRARFLRYSDVAILSLIHVFIGYCTYLCNQQKLGSLRHD






IDIEAPQEEYYSNNEKGTTDNIKYGRRCVIGTVALYLLICT






GIIIYTRTATAQVVWRLPPLVVPVEESEVIFWDCWAPEEPA






CQDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKA






TRQCRRGRIWRRWNETITGPLGCANNTCYNISVIVPDYQC






YLDRVDTWLQGKINISLCLTGGKMLYNRYTKQLSYCTDP






LQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQRAY






YNSCRWESTDVKFHCQRTQSQPGSWLRAISSWRQRNRW






EWRPDFESEKVKVSLQCNSTKNLTFAMRSSGDYGDVTGA






WIEFGCHRNKSRLHTEARFRIRCRWNVGDNTSLIDTCGKT






QNIAGANPVDCTMYVNRMYNCSLQNGFTMKVDDLIMHF






NMTKAVEMYNIAGNWSCTSNLPPTWGYINCNCTNSSDSN






KMACPSSQGILRNWYNPVAGLRQSLEKYQVVKQPDYLV






VPGEVMEYKPRRKRAAIHVMLALATILSMAGAGTGATAI






GMVTQYHQVLATHQEAIEKVTEALKVNNLRLITLEHQVL






VIGLKVEAMEKFRYTAFAMQELGCNQNQFFCKVPPELW






KRYNMTINQTIWNHGNITLGEWYNQTKGLQQKFYEIIMDI






EQNSVQGKKGIQQLQEWEDWIGWIGNIPQYLKGLLGGIL






GIGLGVLLLILCLPTLVDCIRNCIHKILGYTVIAMPEVEGEE






IQPQMELRRNGRQCGMSEKEEE










SEQ ID NO: 388









MNEEGPLNPGINPFRVPGITETEKQDYCNMLQPKLQALRN






EIQEVKLEEGNAGKFRRARFLRYSDETILSLIHLFIGYCTYL






LNRKELGSLRHDIDIEAPQEECYSSREQSITDNIKYGKRCFI






GTAGLYLLLFIGVGIYLGTAKAQVVWRLPPLVVPVEESEII






FWDCWAPEEPACQDFLGAMIHLKASTNISIQEGPTLGNW






AKEIWGTLFKKATRQCRRGRIWKRWNETISGPLGCANNT






CYNISVIVPDYQCYLDRVDTWLQGKVNVSLCLTGGKILY






NKYTKQLSYCTDPLQIPLISYTFGPNQTCMWDTSQIQDPEI






PKCGWWNQIAYYNSCRWESTDVKFHCQRTQSQPGLWLR






AISSWKQRNRWEWRPDFESEKAKVSLQCNSTKNLTFAMR






SSGDYGEVTGAWIEFGCHRNKSKLHTEARFRIRCRWNVG






DNTSLIDTCGETQNVSGANPVDCTMYANRMYNCSLQNGF






TMKVDDLIMHFNMTKAVEMYDIAGNWSCTSDLPPTWGY






MNCNCTNSSSTNSVKMACPKNQGILRNWYNPVAGLRQS






LEKYQVVKQPDYLVVPGEVMEYKPRRKRAAIHVMLALA






TVLSMAGAGTGATAIGMVTQYHQVLATHQETIEKVTEAL






KINNLRLVTLEHQVLVIGLKVEAMEKRLYTAFAMQELGC






NQNQFFCKVPPELWKRYNMTINQTIWNHGNITLGEWYNQ






TKELQQKFYEIIMNIEQNNVQVKKGLQQLQEWEDWVGWI






GNIPQYLKGLLGGILGIGIGVLLLILCLPTLVDCIRNCISKVL






GYTVIAMPEIGDEEETVQMELRKNGRQCGMSEKEEE










SEQ ID NO: 389









MNEEGPLNPGINPFRVPGITETEKQDYCNMLQPKLQALRN






EIQEVKLEEGNAGKFRRARFLRYSDETILSLIHLFIGYCTYL






LNRKELGSLRHDIDIEAPQEECYSSREQSITDNIKYGKRCFI






GTAGLYLLLFIGVGIYLGTAKAQVVWRLPPLVVPVEESEII






FWDCWAPEEPACQDFLGAMIHLKASTNISIQEGPTLGNW






AKEIWGTLFKKATRQCRRGRIWKRWNETISGPLGCANNT






CYNISVIVPDYQCYLDRVDTWLQGKVNVSLCLTGGKILY






NKYTKQLSYCTDPLQIPLISYTFGPNQTCMWDTSQIQDPEI






PKCGWWNQIAYYNSCRWESTDVKFHCQRTQSQPGLWLR






AISSWKQRNRWEWRPDFESEKAKVSLQCNSTKNLTFAMR






SSGDYGEVTGAWIEFGCHRNKSKLHTEARFRIRCRWNVG






DNTSLIDTCGETQNVSGANPVDCTMYANRMYNCSLQNGF






TMKVDDLIMHFNMTKAVEMYDIAGNWSCTSDLPPTWGY






MNCNCTNSSSTNSVKMACPKNQGILRNWYNPVAGLRQS






LEKYQVVKQPDYLVVPGEVMEYKPRRKRAAIHVMLALA






TVLSMAGAGTGATAIGMVTQYHQVLATHQETIEKVTEAL






KINNLRLVTLEHQVLVIGLKVEAMEKFRYTAFAMQELGC






NQNQFFCKVPPELWKRYNMTINQTIWNHGNITLGEWYNQ






TKELQQKFYEIIMNIEQNNVQVKKGLQQLQEWEDWVGWI






GNIPQYLKGLLGGILGIGIGVLLLILCLPTLVDCIRNCISKVL






GYTVIAMPEIGDEEETVQMELRKNGRQCGMSEKEEE










SEQ ID NO: 390









<SEQ ID NO: 390; PRT; Artificial sequence>






MAEGFAANRQWIGPEEAEELLDFDIATQMNEEGPLNPGIN






PFRVPGITEIEKRDYCKILQPKLQDLKNEIQEVKLEEGNAG






KFRRARFLRYSDENILSLIHLFIGYCTYLCRKNELGSLRHDI






DIDEHQEEYYTNIEKGTTANIKYGRRCLIGTAALYLLFIGIII






YTQTTKAQVVWRLPPFVVPVEESEIIFWDCWAPEEPACQD






FLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKATRQ






CRRGRVWRRWNETITGPSGCANNTCYNISVIVPDYQCYL






DRVDTWLQGKVNISLCLTGGKMLYNKYTKQLSYCTDPL






QIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQAAYY






NSCRWESTDVKFHCQRTQSLPGTWLRTISSWRPKNRWEW






RPDFESEKVKVSLQCNSTNNLTFAMRSSGDYGEVTGAWI






EFGCHRKKSKLHSEARFRIRCRWDKGDNTSLIDTCGKTQN






VLGANPVDCTMYANRMYNCSLQNGFTMKIDDLVMHFN






MTKAVEMYNIAGNWSCTSDLPPTWGYMNCNCTNSSSTSS






SSGNKMACPGDKGILRNWYNPVAGLRQSLEKYQVVKQP






DYLVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATHQEAIEKVTEALKINNLRLVTLE






HQVLVIGLKVEAMEKRLYTAFAMQELGCNQNQFFCKVPP






VLWERYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEI






IMDIEQNNVQGKKGLQQLQKWEDWVGWIGNIPKYLKGL






LGGILGIGLGVILLILCLPTLVDCVRNCIHKILGYTVIAMPE






VEEEEIQPQMELRRNGRQCGISEKEEE










SEQ ID NO: 391









MAEGFAANRQWIGPEEAEELLDFDIATQMNEEGPLNPGIN






PFRVPGITEIEKRDYCKILQPKLQDLKNEIQEVKLEEGNAG






KFRRARFLRYSDENILSLIHLFIGYCTYLCRKNELGSLRHDI






DIDEHQEEYYTNIEKGTTANIKYGRRCLIGTAALYLLFIGIII






YTQTTKAQVVWRLPPFVVPVEESEIIFWDCWAPEEPACQD






FLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKKATRQ






CRRGRVWRRWNETITGPSGCANNTCYNISVIVPDYQCYL






DRVDTWLQGKVNISLCLTGGKMLYNKYTKQLSYCTDPL






QIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQAAYY






NSCRWESTDVKFHCQRTQSLPGTWLRTISSWRPKNRWEW






RPDFESEKVKVSLQCNSTNNLTFAMRSSGDYGEVTGAWI






EFGCHRKKSKLHSEARFRIRCRWDKGDNTSLIDTCGKTQN






VLGANPVDCTMYANRMYNCSLQNGFTMKIDDLVMHFN






MTKAVEMYNIAGNWSCTSDLPPTWGYMNCNCTNSSSTSS






SSGNKMACPGDKGILRNWYNPVAGLRQSLEKYQVVKQP






DYLVVPGEVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATHQEAIEKVTEALKINNLRLVTLE






HQVLVIGLKVEAMEKFRYTAFAMQELGCNQNQFFCKVPP






VLWERYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEI






IMDIEQNNVQGKKGLQQLQKWEDWVGWIGNIPKYLKGL






LGGILGIGLGVILLILCLPTLVDCVRNCIHKILGYTVIAMPE






VEEEEIQPQMELRRNGRQCGISEKEEE










SEQ ID NO: 392









MAEGFAVNRQWIGPEEAEELLDFDIATQMNEEGPLNPGIN






LFRVPGITETEKQEYCNILQPKLQDLRNEIQEVKLEEGNAG






KFRRARFLRYSDETILSLIHLFIGYCTYLCKRNKLGTLVHN






IDIEAPQEECYSNRERGTTVNIKYSRRCCIGTTALYLLLLT






GIIIYTQTTQAQVVWRLPPLVVPVEESEIIFWDCWAPEEPA






CQDFLGAMIYLKASTNISIQEGPTLGNWAREIWGTLFKKA






TRQCRRGRIWRRWNETITGPSGCANNTCYNISVIVPDYQC






YLDRVDTWLQGKVNISLCLTGGKMLYNKDTKQLSYCTD






PLQIPLINYTFGPNQTCMWNTSQIQDSDIPKCGWWNQIAY






YNSCRWEQTDVKFHCQRTQSQPGTWLRTISSWKQKNRW






EWRPDFESEKVRVSLQCNTTKNLTFAMRSSGDYGEVTGA






WIEFGCHRNKSKLHSDARFRIRCRWNVGDNTSLIDTCGN






DPNVSGANPVDCTMYANRMYNCSLQNGFTMKVDDLIMH






FNMTKAVEMYNIAGNWSCTSDLPSTWGYMNCNCTNSSS






TDSNKMACPKRQGILRNWYNPVAGLRQSLEKYQVVKQP






DYLVVPREVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATHQETIEKITEALKVNNLRLVTLE






HQVLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKVPP






ELWQRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEI






IMDMEQNNVQGRKGLQQLQEWEDWVGWLGNIPRYLKG






LLGGILGIGLGVLLLILCLPTLVDCIRNCISKVLGYTVIAMP






EVEEEEIQPPMELRRNGRQCDMSEKEEE










SEQ ID NO: 393









MAEGFAVNRQWIGPEEAEELLDFDIATQMNEEGPLNPGIN






LFRVPGITETEKQEYCNILQPKLQDLRNEIQEVKLEEGNAG






KFRRARFLRYSDETILSLIHLFIGYCTYLCKRNKLGTLVHN






IDIEAPQEECYSNRERGTTVNIKYSRRCCIGTTALYLLLLT






GIIIYTQTTQAQVVWRLPPLVVPVEESEIIFWDCWAPEEPA






CQDFLGAMIYLKASTNISIQEGPTLGNWAREIWGTLFKKA






TRQCRRGRIWRRWNETITGPSGCANNTCYNISVIVPDYQC






YLDRVDTWLQGKVNISLCLTGGKMLYNKDTKQLSYCTD






PLQIPLINYTFGPNQTCMWNTSQIQDSDIPKCGWWNQIAY






YNSCRWEQTDVKFHCQRTQSQPGTWLRTISSWKQKNRW






EWRPDFESEKVRVSLQCNTTKNLTFAMRSSGDYGEVTGA






WIEFGCHRNKSKLHSDARFRIRCRWNVGDNTSLIDTCGN






DPNVSGANPVDCTMYANRMYNCSLQNGFTMKVDDLIMH






FNMTKAVEMYNIAGNWSCTSDLPSTWGYMNCNCTNSSS






TDSNKMACPKRQGILRNWYNPVAGLRQSLEKYQVVKQP






DYLVVPREVMEYKPRRKRAAIHVMLALATVLSMAGAGT






GATAIGMVTQYHQVLATHQETIEKITEALKVNNLRLVTLE






HQVLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKVPPE






LWQRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEII






MDMEQNNVQGRKGLQQLQEWEDWVGWLGNIPRYLKGL






LGGILGIGLGVLLLILCLPTLVDCIRNCISKVLGYTVIAMPE






VEEEEIQPPMELRRNGRQCDMSEKEEE










SEQ ID NO: 394









MAAGFSQNRQWIGPEEAEELLDFDIATQINEEGPLNPGVN






PFRVPGITDTEKQDYCKILQPKLQELREEINEVKLDEDNAG






KFRRVRYLRYADETVLSLIYALVGYLRYLGNRNKLGSLR






HDIDIEVSAKEQFDKKEKGTTINQKYCTRCCVGISVLYLIL






FIIIVAVTGSQAQVVWRHPPLVVPVEETEIIFWDCWAPEEP






ACQDFLGTMVQLKASINIGIQEGPTLGHWAREIWSTLF






KKATRQCRRGRVWRRWNETITGPLGCANNTCYNISVVVP






DYQCYVDRVDTWLQGRINISLCLTGGKMLYNKDTQQLS






YCTEPLQIPLINYTFGPNQTCMWNTSLIEDSEIPKCGWWN






QAAYYNSCKWEQTDVKFQCLRTQSQPGNWLRTISSWKQ






SNRWIWRPDFESDKVKISLQCNSTKNLTFAMRSSGDYGEI






TGAWIEFGCYRNKSKSHGEARFRIRCRWNEGTNTSLIDTC






GSTPNVKGANPVDCTMKANTMYNCSLQNGFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCNSDIPPGWGYMNCNCTNST






SSSFPTCPKMACPRERGILRNWYNPIAGLRHALQKYQVVK






QPAYLLVPEEVMEYKPSQKRAAIHIILALATVLSIAGAGTG






ATAIGMVTQYHQVLATHQKAIDQITEALKINNLRLVTLEH






QVLVIGLKVEAIEKRIYTAFAMQELGCNQNQFFCKIPPEL






WIRYNMSINQTIWNHGNISLRDWYNNTQQLQKKFYEIIYD






IEQNNVQGKQGLQQLQKWETWVSWIGKIPQYLKGLFGSI






LGIGLGILLMILCLPTLVDCMRNCLNRVLGYTVIAMPTIDD






EGTDFPVELRRNGGQCGMSEKEEE










SEQ ID NO: 395









MAAGFSQNRQWIGPEEAEELLDFDIATQINEEGPLNPGVN






PFRVPGITDTEKQDYCKILQPKLQELREEINEVKLDEDNAG






KFRRVRYLRYADETVLSLIYALVGYLRYLGNRNKLGSLR






HDIDIEVSAKEQFDKKEKGTTINQKYCTRCCVGISVLYLIL






FIIIVAVTGSQAQVVWRHPPLVVPVEETEIIFWDCWAPEEP






ACQDFLGTMVQLKASINIGIQEGPTLGHWAREIWSTLFKK






ATRQCRRGRVWRRWNETITGPLGCANNTCYNISVVVPDY






QCYVDRVDTWLQGRINISLCLTGGKMLYNKDTQQLSYCT






EPLQIPLINYTFGPNQTCMWNTSLIEDSEIPKCGWWNQAA






YYNSCKWEQTDVKFQCLRTQSQPGNWLRTISSWKQSNR






WIWRPDFESDKVKISLQCNSTKNLTFAMRSSGDYGEITGA






WIEFGCYRNKSKSHGEARFRIRCRWNEGTNTSLIDTCGST






PNVKGANPVDCTMKANTMYNCSLQNGFTMKIEDLIVHFN






MTKAVEMYNIAGNWSCNSDIPPGWGYMNCNCTNSTSSSF






PTCPKMACPRERGILRNWYNPIAGLRHALQKYQVVKQPA






YLLVPEEVMEYKPSQKRAAIHIILALATVLSIAGAGTGATA






IGMVTQYHQVLATHQKAIDQITEALKINNLRLVTLEHQVL






VIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIPPELWIR






YNMSINQTIWNHGNISLRDWYNNTQQLQKKFYEIIYDIEQ






NNVQGKQGLQQLQKWETWVSWIGKIPQYLKGLFGSILGI






GLGILLMILCLPTLVDCMRNCLNRVLGYTVIAMPTIDDEG






TDFPVELRRNGGQCGMSEKEEE










SEQ ID NO: 396









MAAGFTQNRQWIGPEEAEELLDFDIVTQINEEGPLNPGVN






PFRVPAITDTEKQDYCKILQPKLQELREEIKETKLDESNAG






KFRRVRYLRYADETALSLIYALVGYLRYLLERRKLGSLR






HDVDIEVSAKEQFNKKEKGTTVNQNYCTKCCVGISVLYFI






LFLILVAVTRSQAQVVWRLPPLVVPVEETEIIFWDCWAPE






EPACQDFLGTMVQLKASINISIQEGPTLGHWAREILETLF






KKATRQCRRGRVWKRWNETITGPLGCANNTCYNISVVVP






DYQCYVDRVDTWLQGRINISLCLTGGKMLYNKDTQQLS






YCTEPLQIPLINYTFGPNQTCMWNTSLIEDSEIPKCGWWN






QAAYYNSCKWEQTDVKFQCQRTQSQPGTWLRAIASWKQ






ANRWIWRPDFESDKVKISLQCNSTKNLTFAMRSSSDYGEI






TGAWIEFGCYRNKSKFHDEARFRIRCRWNEGTNTSLIDTC






GDNPNVTGANPVDCTMRANIMYNCSLQNGFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TDNTKMTCPENQGILRNWYNPVAGLRQALMKYQVVKQP






EYLIVPEEVMQYKSKQKRAAIHIMLALATVLSMAGAGTG






ATAIGMVTQYHQVLATHQQALDKITEALKINNLRLITLEH






QVLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKIPPSL






WSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYEII






MDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGLL






GSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMPEI






DDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 397









MAAGFTQNRQWIGPEEAEELLDFDIVTQINEEGPLNPGVN






PFRVPAITDTEKQDYCKILQPKLQELREEIKETKLDESNAG






KFRRVRYLRYADETALSLIYALVGYLRYLLERRKLGSLR






HDVDIEVSAKEQFNKKEKGTTVNQNYCTKCCVGISVLYFI






LFLILVAVTRSQAQVVWRLPPLVVPVEETEIIFWDCWAPE






EPACQDFLGTMVQLKASINISIQEGPTLGHWAREILETLF






KKATRQCRRGRVWKRWNETITGPLGCANNTCYNISVVVP






DYQCYVDRVDTWLQGRINISLCLTGGKMLYNKDTQQLS






YCTEPLQIPLINYTFGPNQTCMWNTSLIEDSEIPKCGWWN






QAAYYNSCKWEQTDVKFQCQRTQSQPGTWLRAIASWKQ






ANRWIWRPDFESDKVKISLQCNSTKNLTFAMRSSSDYGEI






TGAWIEFGCYRNKSKFHDEARFRIRCRWNEGTNTSLIDTC






GDNPNVTGANPVDCTMRANIMYNCSLQNGFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TDNTKMTCPENQGILRNWYNPVAGLRQALMKYQVVKQP






EYLIVPEEVMQYKSKQKRAAIHIMLALATVLSMAGAGTG






ATAIGMVTQYHQVLATHQQALDKITEALKINNLRLITLEH






QVLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIPPSL






WSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYEII






MDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGLL






GSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMPEI






DDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 398









MAEGFCQNRQWIGPEEAEELLDFDIATQVSEEGPLNPGIN






PFRQPGLTDGEKEEYCKILQPRLQALREEYKEGSLNSESA






GKYRRVRYLRYSDLRVLSLLYLFIGYLAFFVRKRGLGKQ






RQDIDIESKGTEEKFSKNEKGQTVNIRNCRILTIAICSFYIFL






FIGIGIYAGKGEAQVIWRLPPLVVPVEDSEIIFWDCWAPEE






PACQDFLGAMMHLKASTNISIQEGPTLGKWAKEIWATLF






KKATRQCRRGKVWRKWNETITGPKGCANNTCYNVTVSIP






DYQCYLDRVDTWLQGKVNISLCLTGGKMLYNKETKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSLIKNPDIPKCGWWN






QAVYYNSCRWEKADVQFQCQRTQSQPGTWLRKISSWKQ






KNRWEWRPDFESERVKISLQCNSTKNLTFAMRSSSDYSD






VVGAWIEFGCHRNKSRRHTDARFRIRCKWNVGSNTSLID






TCGKENITGANPVDCTMTAKTLYNCSLQEGFTMKIEDLIM






HFNMTKAVEMYEIAGNWSCKSDLPTDWGYMKCNCTSRN






ESEKMKCPAKDGILRNWYNPVAGLRQALDKYQVVKQPD






YIVVPEEVLNYQSRQKRAAIHIMLALATVLSIAGAGTGAT






AIGMVTQYHQVLATHQEALDKITEALKINNLRLVTLEHQ






VLVIGLKVEATEKRLYTAFAMQELGCNQNQFFCKIPCEL






WMRYNLTLNQTIWNHGNVTLQDWYNQTKQLQQKFYEII






MDIEQNNVQGKKGIQQLQSWEYWTGWMGKIPQYLKGLL






GGVLGIGLGILLLILCLPTLLDCMRNCINKVMGYTVIVMPE






IDDEELSQNMELRRNGRQCGMSEKEEE










SEQ ID NO: 399









MAEGFCQNRQWIGPEEAEELLDFDIATQVSEEGPLNPGIN






PFRQPGLTDGEKEEYCKILQPRLQALREEYKEGSLNSESA






GKYRRVRYLRYSDLRVLSLLYLFIGYLAFFVRKRGLGKQ






RQDIDIESKGTEEKFSKNEKGQTVNIRNCRILTIAICSFYIFL






FIGIGIYAGKGEAQVIWRLPPLVVPVEDSEIIFWDCWAPEE






PACQDFLGAMMHLKASTNISIQEGPTLGKWAKEIWATLF






KKATRQCRRGKVWRKWNETITGPKGCANNTCYNVTVSIP






DYQCYLDRVDTWLQGKVNISLCLTGGKMLYNKETKQLS






YCTDPLQIPLINYTFGPNQTCMWNTSLIKNPDIPKCGWWN






QAVYYNSCRWEKADVQFQCQRTQSQPGTWLRKISSWKQ






KNRWEWRPDFESERVKISLQCNSTKNLTFAMRSSSDYSD






VVGAWIEFGCHRNKSRRHTDARFRIRCKWNVGSNTSLID






TCGKENITGANPVDCTMTAKTLYNCSLQEGFTMKIEDLIM






HFNMTKAVEMYEIAGNWSCKSDLPTDWGYMKCNCTSRN






ESEKMKCPAKDGILRNWYNPVAGLRQALDKYQVVKQPD






YIVVPEEVLNYQSRQKRAAIHIMLALATVLSIAGAGTGAT






AIGMVTQYHQVLATHQEALDKITEALKINNLRLVTLEHQ






VLVIGLKVEATEKFRYTAFAMQELGCNQNQFFCKIPCEL






WMRYNLTLNQTIWNHGNVTLQDWYNQTKQLQQKFYEII






MDIEQNNVQGKKGIQQLQSWEYWTGWMGKIPQYLKGLL






GGVLGIGLGILLLILCLPTLLDCMRNCINKVMGYTVIVMPE






IDDEELSQNMELRRNGRQCGMSEKEEE










SEQ ID NO: 400









MAEGGFAQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITPQEKDNYCTILQPRLQELKREIKEVKIEEEN






AGKFRRARFLRYSDENVLSIVYLLTGYLRYLIDRKSLGSL






RHDIDIEVPQKEQYSNNEKGNTVNRKYGRICCISTLFLYLL






LFAGIGVWYGTTAQVVWRLPPLVVPIDDTEIIFWDCWAPE






EPACQDFLGAMIHLKANINISIQEGPTLGNWAREIWSTLFK






KATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVIPDY






QCYVDRVDTWLQGKVNISLCLTGGKMLFNKETKQLSYC






TDPLQIPLINYTFGPNQTCEWNTSLIKDPEIPKCGWWNQN






AYYNSCKWEQTDVKFQCQRIQSQPGSWIRAISSWRQRNR






WEWRPDFESEKVKISLQCNSTRNLTFAMRSSSDYYDVQG






AWIEFGCYRNKSIRHTGTRFRIRCRWNEGKNMSLIDTCGT






DPNVTRANPVNCTLKTNTMYNCTLQDSFTMKIEDLIVHF






NMSKAVEMYNIAGNWSCTSDLPTGWGYMKCNCTSTNNT






GKMKCPEPEGILRNWYNPVAGLRQALMKYQVVKQPEYL






IVPEEVMKYKSKQKRAAIHIMLALATVLSIAGAGTGATAI






GMVTQYHQVLATHQQALEKITEALKINNLRLVTLEHQVL






MIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKIPLNLWT






MYNMTINQTLWNHGNITLGDWYNQTKGLQEKFYEIIMDL






EQNNVQGKLGIQQLQKWENWVGWIGKIPQYLKGLLGSV






LGIGVGILLLIICLPTLVDCIRNCINKVLGYSVIAMPELDDE






EVSMELRRNGRQCGMSEKEEE










SEQ ID NO: 401









MAEGGFAQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITPQEKDNYCTILQPRLQELKREIKEVKIEEEN






AGKFRRARFLRYSDENVLSIVYLLTGYLRYLIDRKSLGSL






RHDIDIEVPQKEQYSNNEKGNTVNRKYGRICCISTLFLYLL






LFAGIGVWYGTTAQVVWRLPPLVVPIDDTEIIFWDCWAPE






EPACQDFLGAMIHLKANINISIQEGPTLGNWAREIWSTLFK






KATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVIPDY






QCYVDRVDTWLQGKVNISLCLTGGKMLFNKETKQLSYC






TDPLQIPLINYTFGPNQTCEWNTSLIKDPEIPKCGWWNQN






AYYNSCKWEQTDVKFQCQRIQSQPGSWIRAISSWRQRNR






WEWRPDFESEKVKISLQCNSTRNLTFAMRSSSDYYDVQG






AWIEFGCYRNKSIRHTGTRFRIRCRWNEGKNMSLIDTCGT






DPNVTRANPVNCTLKTNTMYNCTLQDSFTMKIEDLIVHF






NMSKAVEMYNIAGNWSCTSDLPTGWGYMKCNCTSTNNT






GKMKCPEPEGILRNWYNPVAGLRQALMKYQVVKQPEYL






IVPEEVMKYKSKQKRAAIHIMLALATVLSIAGAGTGATAI






GMVTQYHQVLATHQQALEKITEALKINNLRLVTLEHQVL






MIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIPLNLWT






MYNMTINQTLWNHGNITLGDWYNQTKGLQEKFYEIIMDL






EQNNVQGKLGIQQLQKWENWVGWIGKIPQYLKGLLGSV






LGIGVGILLLIICLPTLVDCIRNCINKVLGYSVIAMPELDDE






EVSMELRRNGRQCGMSEKEEE










SEQ ID NO: 402









MAEGGFTQNHQWIGPEEAEELLDFDIAIQMNEEGPLNPGV






NPFRVPGITSQEKDDYCKILQTKLQELKNEVKEVKIEEGN






AGKFRRARFLRYSDENVLSIVYLLIGYLRYLIDHRSLGSLR






HDIDIEAPQEEHYNNSEKGTTLNIKYGRRCCISTFIMYLILF






AGVGIWFGARAQVVWRLPPLVVPVDDTEIIFWDCWAPEE






PACQDFLGTMIHLKAKTNISIQEGPTLGNWAREIWATLFK






KATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVVPD






YQCYLDRVDTWLQGKLNISLCLTGGKMLYNKVTKQLSY






CTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQ






MAYYNSCKWEEAKVKFHCQRTQSQPGSWHRAISSWKQR






NRWEWRPDFKSKKVKISLQCNSTKNLTFAMRSSSDYYDV






QGAWIEFGCHRNKSKGFSEARFRTRCKWNEGNNISLIDTC






GTNPNVTGANPVDCTMKANIMYNCSLQDSFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TDNSKTKMACPRNQGILRNWYNPVAGLRQALIKYQVVK






QPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGAG






TGATAIGMVTQYHQVLATHQHALDKITEALKINNLRLITL






VHQVLVIGLKVRAIEKRLYTAFAMQELGCNQNQFFCKIPP






SLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYE






IIMDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGL






LGSVLGIGLGILLLLICLPTLVDCIRNCTHKILGYTVIAMPEI






DDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 403









MAEGGFTQNHQWIGPEEAEELLDFDIAIQMNEEGPLNPGV






NPFRVPGITSQEKDDYCKILQTKLQELKNEVKEVKIEEGN






AGKFRRARFLRYSDENVLSIVYLLIGYLRYLIDHRSLGSLR






HDIDIEAPQEEHYNNSEKGTTLNIKYGRRCCISTFIMYLILF






AGVGIWFGARAQVVWRLPPLVVPVDDTEIIFWDCWAPEE






PACQDFLGTMIHLKAKTNISIQEGPTLGNWAREIWATLFK






KATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVVPD






YQCYLDRVDTWLQGKLNISLCLTGGKMLYNKVTKQLSY






CTDPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQ






MAYYNSCKWEEAKVKFHCQRTQSQPGSWHRAISSWKQR






NRWEWRPDFKSKKVKISLQCNSTKNLTFAMRSSSDYYDV






QGAWIEFGCHRNKSKGFSEARFRTRCKWNEGNNISLIDTC






GTNPNVTGANPVDCTMKANIMYNCSLQDSFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TDNSKTKMACPRNQGILRNWYNPVAGLRQALIKYQVVK






QPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGAG






TGATAIGMVTQYHQVLATHQHALDKITEALKINNLRLITL






VHQVLVIGLKVRAIEKPRYTAFAMQELGCNQNQFFCKIPP






SLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYE






IIMDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGL






LGSVLGIGLGILLLLICLPTLVDCIRNCTHKILGYTVIAMPEI






DDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 404









MAEGFAANRQWIGPEEAEELLDFDKATQMNEEGPLNPGV






NPFRVPAVTEADKQEYCKILQPRLQEIRNEIQEVKLEEGN






AGKFRRARFLRYSDESILSLIHLFIGYCTYLVNRRRLGSLR






HDINIEAPQEEQYSSREQGTTENIKYGRRCLIGTASLYLLLF






IGVAIYLGTTNAQIVWRLPPLVVPVEESEIIFWDCWAPEEP






ACQDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKK






ATRHCRRNKIWKRWNETITGPVGCANNTCYNISVIIPDYQ






CYLDRVDTWLQGKVNISLCLTGGKMLYNRDTKQLSYCT






DPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQIA






YYNSCRWESTNVKFYCQRTQSQPGTWIRTISSWRQKNRW






EWRPDFESEKVKISLQCNSTHNLTFAMRSSGDYGEVMGA






WIEFGCHRNKSRFHTEARFRIRCRWNVGDNTSLIDTCGKN






LNVSGANPVDCTMYANKMYNCSLQNGFTMKVDDLIMHF






NMTKAVEMYNIAGNWSCKSDLPQNWGYMNCNCTNGTS






NDNKMACPEDKGILRNWYNPVAGLRQALEKYQVVKQPE






YIVVPTEVMTYKYKQKRAAIHIMLALATVLSIAGAGTGAT






AIGMVTQYQQVLATHQEALDKITEALKINNLRLVTLEHQ






MLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCEIPKELW






LRYNMTLNQTIWNHGNITLGEWYNQTKYLQQKFYEIIMD






IEQNNVQGKQGLQKLQNWQDWMGWIGKIPQYLKGLLGG






ILGIGLGILLLILCLPTLVDCIRNCISKVLGYTVIAMPEIDDE






EETVQMELRKNGRQCGMSEKEEE










SEQ ID NO: 405









MAEGFAANRQWIGPEEAEELLDFDKATQMNEEGPLNPGV






NPFRVPAVTEADKQEYCKILQPRLQEIRNEIQEVKLEEGN






AGKFRRARFLRYSDESILSLIHLFIGYCTYLVNRRRLGSLR






HDINIEAPQEEQYSSREQGTTENIKYGRRCLIGTASLYLLLF






IGVAIYLGTTNAQIVWRLPPLVVPVEESEIIFWDCWAPEEP






ACQDFLGAMIHLKASTNISIQEGPTLGNWAREIWGTLFKK






ATRHCRRNKIWKRWNETITGPVGCANNTCYNISVIIPDYQ






CYLDRVDTWLQGKVNISLCLTGGKMLYNRDTKQLSYCT






DPLQIPLINYTFGPNQTCMWNTSQIQDPEIPKCGWWNQIA






YYNSCRWESTNVKFYCQRTQSQPGTWIRTISSWRQKNRW






EWRPDFESEKVKISLQCNSTHNLTFAMRSSGDYGEVMGA






WIEFGCHRNKSRFHTEARFRIRCRWNVGDNTSLIDTCGKN






LNVSGANPVDCTMYANKMYNCSLQNGFTMKVDDLIMHF






NMTKAVEMYNIAGNWSCKSDLPQNWGYMNCNCTNGTS






NDNKMACPEDKGILRNWYNPVAGLRQALEKYQVVKQPE






YIVVPTEVMTYKYKQKRAAIHIMLALATVLSIAGAGTGAT






AIGMVTQYQQVLATHQEALDKITEALKINNLRLVTLEHQ






MLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCEIPKELW






LRYNMTLNQTIWNHGNITLGEWYNQTKYLQQKFYEIIMD






IEQNNVQGKQGLQKLQNWQDWMGWIGKIPQYLKGLLGG






ILGIGLGILLLILCLPTLVDCIRNCISKVLGYTVIAMPEIDDE






EETVQMELRKNGRQCGMSEKEEE










SEQ ID NO: 406









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKEDYCKILQTKLQELKNEVKGVKIEEG






NAGKFRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIETPQEEHFNNSEKGTTLNTKYGRRCCLSTFIMNLI






LFAGVGIWLGARAQVVWRLPPLVVPVDDTEIIFWDCWAP






EEPACQDFLGTMVQLKASINISIQEGPTLGHWAREILETLF






KKATRQCRRGRVWRRWNETITGPLGCANNACYNISVVVP






DYQCYVDRVDTWLQGRINISLCLTGGKMLYNKDTQQLS






YCTEPLQIPLINYTFGPNQTCMWNTSLIEDSEIPKCGWWN






QAAYYNSCKWEQTDVKFQCQRTQSQPGTWLRAIASWEQ






ANRWIWRPDFESDKVKISLQCNSTKNLTFAMRSSSDYYD






VQGAWIEFGCHRNKSRRHSEARFRIRCKWNEGKNISLIDT






CGTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLI






VHFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTN






GTDDSKTKMACPENQGILRNWYNPVAGLRQALIKYHVV






KQPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGA






GTGATAIGMVTQYHQVLATHQQALDKITEALKINNLRLIT






LEHQVLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKIP






PSLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFY






EIIMDIEQNNVQGKTGIQQLQKWENWMGWIGKIPQYLKG






LLGSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMP






EIDDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 407









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKEDYCKILQTKLQELKNEVKGVKIEEG






NAGKFRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIETPQEEHFNNSEKGTTLNTKYGRRCCLSTFIMNLI






LFAGVGIWLGARAQVVWRLPPLVVPVDDTEIIFWDCWAP






EEPACQDFLGTMVQLKASINISIQEGPTLGHWAREILETLF






KKATRQCRRGRVWRRWNETITGPLGCANNACYNISVVVP






DYQCYVDRVDTWLQGRINISLCLTGGKMLYNKDTQQLS






YCTEPLQIPLINYTFGPNQTCMWNTSLIEDSEIPKCGWWN






QAAYYNSCKWEQTDVKFQCQRTQSQPGTWLRAIASWEQ






ANRWIWRPDFESDKVKISLQCNSTKNLTFAMRSSSDYYD






VQGAWIEFGCHRNKSRRHSEARFRIRCKWNEGKNISLIDT






CGTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLI






VHFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTN






GTDDSKTKMACPENQGILRNWYNPVAGLRQALIKYHVV






KQPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGA






GTGATAIGMVTQYHQVLATHQQALDKITEALKINNLRLIT






LEHQVLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIP






PSLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFY






EIIMDIEQNNVQGKTGIQQLQKWENWMGWIGKIPQYLKG






LLGSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMP






EIDDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 408









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKEDYCKTLQTNLQELKNEVKEVKIEEG






NAGKFRRARFLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIEAPQEEHYNNSEKGTTLNIKYARRCCISTFIMYLIL






FAGVGIWLGARAQVVWRLPPLVVPVDDTEIIFWDCWAPE






EPACQDFLGTMVQLKASINIGIQEGPTLGHWAREIWSTLF






KKATRQCRRGRVWRRWNETITGPLGCANNTCYNISVVVP






DYQCYVDRVDTWLQGRINISLCLTGGKMLYNKDTHQLS






YCTEPLQIPLINCTFGPIQTCMWNTSLIQDPEIPKCGWWTQ






VAYYNNCKWEEANVTFQCHRTQSRSGSWLRTISSWRQR






NRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDYYDV






QGAWIEFGCHRNKSKKHSEARFRIRCKWNEGNNISLIDTC






GTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TDDSKTKMACPENQGILRNWYNPVAGLRQALIKYQVVK






QPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGAG






TGATAIGMVTQYHQVLATHQQALDKITEALKINNLRLITL






EHQVLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKIPP






SLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYE






IIMDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGL






LGSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMPEI






DDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 409









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKEDYCKTLQTNLQELKNEVKEVKIEEG






NAGKFRRARFLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIEAPQEEHYNNSEKGTTLNIKYARRCCISTFIMYLIL






FAGVGIWLGARAQVVWRLPPLVVPVDDTEIIFWDCWAPE






EPACQDFLGTMVQLKASINIGIQEGPTLGHWAREIWSTLF






KKATRQCRRGRVWRRWNETITGPLGCANNTCYNISVVVP






DYQCYVDRVDTWLQGRINISLCLTGGKMLYNKDTHQLS






YCTEPLQIPLINCTFGPIQTCMWNTSLIQDPEIPKCGWWTQ






VAYYNNCKWEEANVTFQCHRTQSRSGSWLRTISSWRQR






NRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDYYDV






QGAWIEFGCHRNKSKKHSEARFRIRCKWNEGNNISLIDTC






GTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TDDSKTKMACPENQGILRNWYNPVAGLRQALIKYQVVK






QPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGAG






TGATAIGMVTQYHQVLATHQQALDKITEALKINNLRLITL






EHQVLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIPP






SLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYE






IIMDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGL






LGSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMPEI






DDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 410









MAEGGFTHNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKDDYCKILQTKLQELKNEVKEVKIEEG






NAGKFRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIETPQEEHYNNSEKGTTLNIKYGRRCCISTFIMYLIL






FAGVGIWLGARAQVVWRLPPLVVPVDDTEIIFWDCWAPE






EPACQDFLGTMIYLKANVNISIQEGPTLGNWAREIWSTLF






KKATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVVP






DYQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQLS






YCTDPLQIPLINYTFGPNQTCMWNTSLIKDSEIPKCGWWN






QVAYYDTCKWEEANVTFQCHRTQSQSGSWIRTISSWKQR






NRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDYYDV






QGAWIEFGCHRNKSKRHSEARFRIRCKWNEGNNISLIDTC






GTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLIV






HFNMTKAVELYNIAGNWSCTSDLPKGWGYMNCNCTNGT






DNSETKMACPKNQGILRNWYNPVAGLRQALIKYQVVKQ






PEYLIVPEEVMQYKFKQKRAAIHIMLALATVLSMAGAGT






GATAIGMVTQYHQVLATHQQALEKITEALKINNLRLITLE






HQVLVIGLRVEAIEKRLYTAFAMQELGCNQNQFFCKIPPS






LWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYEII






MDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGLL






GSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMPEI






DDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 411









MAEGGFTHNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKDDYCKILQTKLQELKNEVKEVKIEEG






NAGKFRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIETPQEEHYNNSEKGTTLNIKYGRRCCISTFIMYLIL






FAGVGIWLGARAQVVWRLPPLVVPVDDTEIIFWDCWAPE






EPACQDFLGTMIYLKANVNISIQEGPTLGNWAREIWSTL






FKKATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVV






PDYQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQL






SYCTDPLQIPLINYTFGPNQTCMWNTSLIKDSEIPKCGWW






NQVAYYDTCKWEEANVTFQCHRTQSQSGSWIRTISSWKQ






RNRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDYYD






VQGAWIEFGCHRNKSKRHSEARFRIRCKWNEGNNISLIDT






CGTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLI






VHFNMTKAVELYNIAGNWSCTSDLPKGWGYMNCNCTN






GTDNSETKMACPKNQGILRNWYNPVAGLRQALIKYQVV






KQPEYLIVPEEVMQYKFKQKRAAIHIMLALATVLSMAGA






GTGATAIGMVTQYHQVLATHQQALEKITEALKINNLRLIT






LEHQVLVIGLRVEAIEKFRYTAFAMQELGCNQNQFFCKIP






PSLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFY






EIIMDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKG






LLGSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAMP






EIDDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 412









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLHPGI






NPFRVPGITPQEKEDYCKILQPKLQELKGEIKEVKLEERNA






GKFRRARYLRYSDENVLSIIYLLTGYFRYLINHRSLGALRH






DIDIEAPQKEQYSNNEKGTTLNIKYGRVCCISTLLLYLLLF






AGLGVWYGTSAQVVWRLPPLVVPVEDTEIIFWDCWAPEE






PACQDFLGTMIHLKANVNISIQEGPTLGNWAREIWSTLFK






KATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVVPD






YQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQLSY






CTDPLQIPLINYTFGPNQTCMWNTSLIKDSEIPKCGWWNQ






VAYYNACKWEEANVTFQCHRTQSQPGSWIRTISSWKQRN






RWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDYYDIQG






AWIEFGYHRNKSKMHSEVRFRIRCKWNEGNNISLIDTCGT






NPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLIVHF






NMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNGTD






ESGTKMACPKSQGILRNWYNPVAGLRQALIKYQVVKQPE






YLIVPEEVMQYKFKQKRAAIHIMLALATVLSMAGAGTGA






TAIGMVTQYHQVLATHQQALDKITQALKINNLRLITLEHQ






VLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKIPPSLW






SMYNMTLNQTIWNHGNISLGNWYNQTKDLQNKFYEIIMD






IEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGLLGSV






LGIGVGILLLIICLPTLVDCVRNCINKVLGYSVIAMPELDDE






EVSMELRRNGRQCGMSEKEEE










SEQ ID NO: 413









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLHPGI






NPFRVPGITPQEKEDYCKILQPKLQELKGEIKEVKLEERNA






GKFRRARYLRYSDENVLSIIYLLTGYFRYLINHRSLGALRH






DIDIEAPQKEQYSNNEKGTTLNIKYGRVCCISTLLLYLLLF






AGLGVWYGTSAQVVWRLPPLVVPVEDTEIIFWDCWAPEE






PACQDFLGTMIHLKANVNISIQEGPTLGNWAREIWSTLFK






KATRQCRRGRIWRRWNETITGPLGCANNTCYNISVVVPD






YQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQLSY






CTDPLQIPLINYTFGPNQTCMWNTSLIKDSEIPKCGWWNQ






VAYYNACKWEEANVTFQCHRTQSQPGSWIRTISSWKQRN






RWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDYYDIQG






AWIEFGYHRNKSKMHSEVRFRIRCKWNEGNNISLIDTCGT






NPNVTGANPVDCTMKANTMYNCSLQDSFTMKIEDLIVHF






NMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNGTD






ESGTKMACPKSQGILRNWYNPVAGLRQALIKYQVVKQPE






YLIVPEEVMQYKFKQKRAAIHIMLALATVLSMAGAGTGA






TAIGMVTQYHQVLATHQQALDKITQALKINNLRLITLEHQ






VLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIPPSLW






SMYNMTLNQTIWNHGNISLGNWYNQTKDLQNKFYEIIMD






IEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKGLLGSV






LGIGVGILLLIICLPTLVDCVRNCINKVLGYSVIAMPELDDE






EVSMELRRNGRQCGMSEKEEE










SEQ ID NO: 414









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKDDYCKVLQTKLQELKNEVKEVKIEEG






NAGKFRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIETPQEEHYNNSEKGTTLNTKYGRRCCLSTFIMYL






VLFAGVGLWLGARAQVVWRLPPLVVPVDDTEIIFWDCW






APEEPACQDFLGTMIYLKANVNISIQEGPTLGNWAREIWS






TLFKKATRQCRRGRIWKRWNETITGPLGCATNTCYNISVV






VPDYQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQ






LSYCTDPLQIPLINYTFGPNQTCMWNTSLIKDSEIPKCGW






WNQVAYYNACKWEEANVTFQCQRTQSQSGSWIRTISSW






KQRNRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDY






YDVQGAWIEFGCHRNKSKKYSEARFRIRCKWNEGKNISLI






DTCGTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIE






DLIVHFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNC






TNGTDNSETKMTCPENQGILRNWYNPVAGLRQALIKYQV






VKQPEYLIVPEEVMQYKFKQKRAAIHIMLALATVLSMAG






AGTGATAIGMVTQYHQVLATHQQALEKITEALKINNLRLI






TLEHQVLVIGLKVEAIEKRLYTAFAMQELGCHQNQFFCKI






PPSLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKF






YEIIMDIEQNNVQGKTGIQQLQKWENWMGWIGKIPQYLK






GLLGSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAM






PEIDDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 415









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKDDYCKVLQTKLQELKNEVKEVKIEEG






NAGKFRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGSL






RHDIDIETPQEEHYNNSEKGTTLNTKYGRRCCLSTFIMYL






VLFAGVGLWLGARAQVVWRLPPLVVPVDDTEIIFWDCW






APEEPACQDFLGTMIYLKANVNISIQEGPTLGNWAREIWS






TLFKKATRQCRRGRIWKRWNETITGPLGCATNTCYNISVV






VPDYQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQ






LSYCTDPLQIPLINYTFGPNQTCMWNTSLIKDSEIPKCGW






WNQVAYYNACKWEEANVTFQCQRTQSQSGSWIRTISSW






KQRNRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDY






YDVQGAWIEFGCHRNKSKKYSEARFRIRCKWNEGKNISLI






DTCGTNPNVTGANPVDCTMKANTMYNCSLQDSFTMKIE






DLIVHFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNC






TNGTDNSETKMTCPENQGILRNWYNPVAGLRQALIKYQV






VKQPEYLIVPEEVMQYKFKQKRAAIHIMLALATVLSMAG






AGTGATAIGMVTQYHQVLATHQQALEKITEALKINNLRLI






TLEHQVLVIGLKVEAIEKFRYTAFAMQELGCHQNQFFCKI






PPSLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKF






YEIIMDIEQNNVQGKTGIQQLQKWENWMGWIGKIPQYLK






GLLGSVLGIGLGILLLLICLPTLVDCIRNCTNKILGYTVIAM






PEIDDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 416









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKDDYCKILQTKLRELKNEVKEVKIEEG






NAGKLRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGS






LRHDIDIETPQEEHYNNSEKGTTLNIKYEGRCCLSTFIMHLI






LFAGVGIWLGARAQVVWRLPPLVVPVDDTEMIFWDCWA






PEEPACQDFLGTMIHLKANVNISIQEGPTLGNWAREIWST






LFKKATRQCRRGKIWRRWNETITGPLGCANNTCYNISVV






VPDYQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQ






LSYCTDPLQIPLINYTFGPNQTCMWNASLIKDSEIPKCGW






WNQAAYYNACKWEEANVTFQCHRTQSQSGSWIRTISSW






RQRNRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDY






YDVQGAWIEFGCHRKKSNKHSEARFRIRCTWNEGNNISLI






DTCGTNPNVTGANPVDCTMKANVMYNCTLQDSFTMKIE






DLIVHFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNC






TNGTDNTKMTCPKNQGILRNWYNPVAGLRQALIKYQVV






KQPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGA






GTGATAIGMVTQYHQVLATHQQALDKITEALKINNLRLIT






LEHQVLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKIP






PSLWSMYNMTLNQTIWNHGNISLGNWYNQTKDLQNKFY






EIIMDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKG






LLGSVLGIGLGILLLLICLPTLVDCIRSCTNRILGYTVIAMPE






IDDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 417









MAEGGFTQNQQWIGPEEAEELLDFDIAVQMNEEGPLNPG






VNPFRVPGITSQEKDDYCKILQTKLRELKNEVKEVKIEEG






NAGKLRRARYLRYSDENVLSIVYLLIGYLRYLIDHRSLGS






LRHDIDIETPQEEHYNNSEKGTTLNIKYEGRCCLSTFIMHLI






LFAGVGIWLGARAQVVWRLPPLVVPVDDTEMIFWDCWA






PEEPACQDFLGTMIHLKANVNISIQEGPTLGNWAREIWST






LFKKATRQCRRGKIWRRWNETITGPLGCANNTCYNISVV






VPDYQCYVDRVDTWLQGKVNISLCLTGGKMLYNKETRQ






LSYCTDPLQIPLINYTFGPNQTCMWNASLIKDSEIPKCGW






WNQAAYYNACKWEEANVTFQCHRTQSQSGSWIRTISSW






RQRNRWEWRPDFESEKVKISLQCNSTKNLTFAMRSSSDY






YDVQGAWIEFGCHRKKSNKHSEARFRIRCTWNEGNNISLI






DTCGTNPNVTGANPVDCTMKANVMYNCTLQDSFTMKIE






DLIVHFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNC






TNGTDNTKMTCPKNQGILRNWYNPVAGLRQALIKYQVV






KQPEYLIVPEEVMQYKVKQKRAAIHIMLALATVLSMAGA






GTGATAIGMVTQYHQVLATHQQALDKITEALKINNLRLIT






LEHQVLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIP






PSLWSMYNMTLNQTIWNHGNISLGNWYNQTKDLQNKFY






EIIMDIEQNNVQGKTGIQQLQKWENWVGWIGKIPQYLKG






LLGSVLGIGLGILLLLICLPTLVDCIRSCTNRILGYTVIAMPE






IDDEEVHLSVELRRNGRQCGISEKEEE










SEQ ID NO: 418









MADGGFSQNQQWIGPEEAEKSLDFDIATQMNEEGPLNPG






VNPFRVPGITPQEKEDYCKILQTRLQELKNELKEVKVEQR






NAGKFRRTRFLRYSDENVLSIVYLLIGYLRYLIDRRSLGSL






RHDIDIKIPQEEHYNNSAKDTTLNIKYERRCCIGTFIMYLIL






FAGIGIWFGAKAQVVWRLPPLVVPVEESEIIFWDCWAPEE






PACQDFLGAMIHLKANTNISIQEGPTLGNWAREIWSTLFK






KATKQCRKGGIWTKWKETITGPLGCANNTCYNISVVVPD






YQCYVDRVDTWLQGKVNISLCLTGGKMLFNKETKQLSY






CTDPLQIPLINYTFGPNQTCMWNTSQIQDSEIPKCGWWNQ






IAYYNSCQWEKTDVKFHCQRTQSQPGSWRRAISSWRQRN






RWEWRPDFESKKVKVSLKCNSTKNLTFAMRSSGDYGEV






TGAWIEFGCHRNKSKLHTEARFRIRCKWNEGNNISLIDTC






GTNPNVAEANPVDCTMKANTMYNCSLQDSFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TTSNGIKMACPKQQGILRNWYNPVAGLRQALIKYQVVKQ






PEYLIVPEEVLQYKFKQKRAAIHIMLALATVLSMAGAGTG






ATAIGVVTQYHQVLATHQQALDKITEALKINNLRLITLEH






QVLVIGLKVEAIEKRLYTAFAMQELGCNQNQFFCKIPLNL






WNMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYEII






MDIEQNNVQGHTGIQQLQKWENWVGWIGKIPQYLKGLIG






SVLGIGLGILLLLICLPTLVDCIRSCTNKMLGYTVIAMPEIG






DEGVHLAMELRRNGRQCGISEKEEE










SEQ ID NO: 419









MADGGFSQNQQWIGPEEAEKSLDFDIATQMNEEGPLNPG






VNPFRVPGITPQEKEDYCKILQTRLQELKNELKEVKVEQR






NAGKFRRTRFLRYSDENVLSIVYLLIGYLRYLIDRRSLGSL






RHDIDIKIPQEEHYNNSAKDTTLNIKYERRCCIGTFIMYLIL






FAGIGIWFGAKAQVVWRLPPLVVPVEESEIIFWDCWAPEE






PACQDFLGAMIHLKANTNISIQEGPTLGNWAREIWSTLFK






KATKQCRKGGIWTKWKETITGPLGCANNTCYNISVVVPD






YQCYVDRVDTWLQGKVNISLCLTGGKMLFNKETKQLSY






CTDPLQIPLINYTFGPNQTCMWNTSQIQDSEIPKCGWWNQ






IAYYNSCQWEKTDVKFHCQRTQSQPGSWRRAISSWRQRN






RWEWRPDFESKKVKVSLKCNSTKNLTFAMRSSGDYGEV






TGAWIEFGCHRNKSKLHTEARFRIRCKWNEGNNISLIDTC






GTNPNVAEANPVDCTMKANTMYNCSLQDSFTMKIEDLIV






HFNMTKAVEMYNIAGNWSCTSDLPKGWGYMNCNCTNG






TTSNGIKMACPKQQGILRNWYNPVAGLRQALIKYQVVKQ






PEYLIVPEEVLQYKFKQKRAAIHIMLALATVLSMAGAGTG






ATAIGVVTQYHQVLATHQQALDKITEALKINNLRLITLEH






QVLVIGLKVEAIEKFRYTAFAMQELGCNQNQFFCKIPLNL






WNMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYEII






MDIEQNNVQGHTGIQQLQKWENWVGWIGKIPQYLKGLIG






SVLGIGLGILLLLICLPTLVDCIRSCTNKMLGYTVIAMPEIG






DEGVHLAMELRRNGRQCGISEKEEE






The invention also encompasses the variants of the above sequences, harbouring the above mentioned mutations, and conferring to said variant a decrease of immunosuppressive properties, a substantial absence of immunosuppressive properties or an absence of immunosuppressive properties.


Advantageously: the mutated feline lentiviral proteins are:


1—SEQ ID NO: 5, SEQ ID NO: 317 to 342 and SEQ ID NO: 374 to 419; these mutated ENV proteins having decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties.


advantageously,


2—SEQ ID NO: 5, SEQ ID NO: 317 to 342 and SEQ ID NO: 374 to 419; these mutated ENV proteins having decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties, and being highly expressed at the plasma membrane,


more advantageously,


3—SEQ ID NO: 5, SEQ ID NO: 317 to 342, and SEQ ID NO: 374 to 419; these mutated ENV proteins having decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties, being highly expressed at the plasma membrane and possessing a medium or high fusogenic activity.


As mentioned above “possessing a medium or high fusogenic activity” means that when the ENV preteins are expressed at a cell plasma membrane, by using appropriate expression vectors the fusogenic activity of said proteins as measured by quantitating the ENV-mediated cell-cell fusion (see Materials and Methods) is:

    • either at a level comparable or higher to the ability of the wild type ENV, i.e. high ability
    • or at a lower level compared to the ability of the wild type ENV, i.e. medium or low ability.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance a nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, as defined above.


In a particular embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule is contained in a vector, said vector comprising means allowing the expression of the mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, as defined above.


In another embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said nucleic acid molecule is comprised in a vector, said nucleic acid molecule being placed under the control of sequences that allow the expression of said mutated lentiviral ENV protein, or variant of said protein, or fragments thereof.


In another embodiment, the invention relates to a pharmaceutical composition, as defined above, in particular as a vaccine, comprising a DNA molecule coding for said mutated feline lentiviral ENV protein or a fragment thereof.


DNA vaccines expressing ENV proteins can be produced as described in Bellier et al. (Vaccine, 2009, 27(42):5772-80).


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said vector is chosen among a canarypox vector, a pox vector, a fowlpox, an adenoviral vector, a lentiviral vector, a measles vector, a Sendaï virus vector, a Cytomegalovirus vector or a Modified Vaccinia Ankara vector.


In another embodiment, the invention concerns a pharmaceutical as defined above, comprising at least one nucleic acid molecule coding for a GAG protein and/or a PRO protein and/or a POL protein of a feline lentivirus, said lentivirus being preferably of the same origin as the mutated lentiviral ENV protein.


GAG expression will produce virus-like particles (VLPs) which are particularly advantageous for a vaccine, in particular if the ENV protein is associated with the VLP (Guerbois et al., Virology, 2009, 388:191-203).


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in the same vector as the one which also contains said at least one nucleic acid molecule coding for a GAG protein and/or a PRO protein and/or a POL protein.


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in the same vector as the one which also contains all said at least one nucleic acid molecule coding for a GAG protein and/or a PRO protein and/or a POL protein.


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in a vector which is different from the at least one vector containing said at least one nucleic acid molecule coding for a GAG protein and/or a PRO protein and/or a POL protein.


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, said nucleic acid molecule coding for a GAG protein, said nucleic acid molecule coding for a PRO protein and said nucleic acid molecule coding for a POL protein, are all contained in vectors which are different from each other.


In another embodiment, the invention concerns a pharmaceutical as defined above, comprising at least one nucleic acid molecule coding for a GAG protein of a feline lentivirus, said lentivirus being preferably of the same origin as the mutated lentiviral ENV protein.


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in the same vector as the one which also contains said at least one nucleic acid molecule coding for a GAG protein.


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in the same vector as the one which also contains a nucleic acid molecule coding for a GAG protein.


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in the same vector as the one which also contains a nucleic acid molecule coding for a GAG protein,


said vector being preferably a canary pox vector (Poulet et al., Veterinary Record, 2003, 153(5):141-145; Vaccari et al., Expert Review of Vaccines, 2010, Vol. 9, No 9: 997-1005).


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in a vector which is different from the at least one vector containing said at least one nucleic acid molecule coding for a GAG protein.


In another embodiment, the invention concerns a pharmaceutical as defined above, wherein said nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, and said nucleic acid molecule coding for a GAG protein are contained in vectors which are different from each other.


In another aspect, the invention concerns a pharmaceutical composition as defined above, in association with at least one antiviral compound, preferably for a simultaneous, separated or sequential use.


The composition according to the invention can also be used in combination with the antiviral compositions listed in Table 1 below:









TABLE 1





Examples of antiviral compounds.
















Intelence ® (TMC 125/etravirine)
Non-nucleoside reverse


Tibotec -
transcriptase inhibitor


Agenerase ®(APV/amprenavir)
Protease inhibitor


GSK -


Aptivus ®(TPV/tipranavir)
Protease inhibitor


Boehringer -


Crixivan ®(IDV/indinavir) MSD -
Protease inhibitor


Invirase ®(SQV/saquinavir) Roche -
Protease inhibitor


Kaletra ®(LPV.r/lopinavir + ritonavir)
Protease inhibitor


Abbott


Norvir ®(ritonavir) Abbott -
Protease inhibitor


Prezista ®(TMC 114/darunavir)
Protease inhibitor


Tibotec/Janssen-Cilag -


Reyataz ®(ATZ/atazanavir) BMS -
Protease inhibitor


Telzir ®(APV/fosamprenavir) GSK -
Protease inhibitor


Viracept ®(nelfinavir) Roche -
Protease inhibitor


Fuzeon ®(T20/enfuvirtide) Roche -
Fusion inhibitor


Celsentri ®(maraviroc) Pfizer -
Entry inhibitor


Isentress ®(MK 0518/raltegravir)
Integrase inhibitor


Merck -


Rescriptor ®(delavirdine) Agouron -
Non-nucleoside reverse



transcriptase inhibitor


Sustiva ®(EFV/efavirenz) BMS -
Non-nucleoside reverse



transcriptase inhibitor


Viramune ®(nevirapine) Boehringer -
Non-nucleoside reverse



transcriptase inhibitor


Combivir ®(Retrovir ® + Epivir ®)
Nucleoside reverse transcriptase


GSK -
inhibitor


Emtriva ®(FTC, emtricitabine) Gilead -
Nucleoside reverse transcriptase



inhibitor


Epivir ®(3TC, lamivudine) GSK -
Nucleoside reverse transcriptase



inhibitor


Kivexa ®(Ziagen ® + Epivir ®)
Nucleoside reverse transcriptase


GSK -
inhibitor


Retrovir ®(AZT/zidovudine) QSK -
Nucleoside reverse transcriptase



inhibitor


Trizivir ®(Retrovir ® + Epivir ® +
Nucleoside reverse transcriptase


Ziagen ®) GSK -
inhibitor


Videx ®(ddI/didanosine) BMS -
Nucleoside reverse transcriptase



inhibitor


Viread ®(TDF/tenofovir) Gilead -
Nucleoside reverse transcriptase



inhibitor


Zerit ®(d4T/stavudine) BMS -
Nucleoside reverse transcriptase



inhibitor


Ziagen ®(ABC/abacavir) GSK -
Nucleoside reverse transcriptase



inhibitor


Truvada ®(Emtriva ® + Viread ®)
Nucleoside reverse transcriptase


Gilead -
inhibitor


Atripla ®(Sustiva ® + Emtriva ® +
Nucleoside and non-nucleoside


Viread ®) BMS/GILEAD
reverse transcriptase inhibitor









In another aspect, the invention concerns a pharmaceutical composition as defined above, for its use for stimulating an immune response in a host organism.


In another aspect, the invention concerns a pharmaceutical composition as defined above, for its use for the prevention or the treatment of lentiviral infection, preferably FIV infection.


In another aspect, the invention concerns a pharmaceutical composition as defined above, for its use as a vaccine, in particular against FIV infection.


As mentioned above, the pharmaceutical composition according to the invention encompasses a vaccine, comprising as active substance, a protein as defined above, or a nucleic acid molecule as defined above, or a vector as defined above, or a combination thereof.


In a particular embodiment, the pharmaceutical composition according to the invention encompasses a vaccine as defined above, comprising a nucleic acid coding for a GAG protein as defined above and a nucleic acid coding for a mutated ENV protein as defined above, which are contained in a same vector, said same vector being preferably a canary pox vector.


In a particular embodiment, the pharmaceutical composition according to the invention encompasses a vaccine, comprising a GAG protein as defined above and a mutated ENV protein as defined above.


In a particular embodiment, the pharmaceutical composition according to the invention encompasses a vaccine, comprising a GAG protein as defined above and a mutated ENV protein as defined above which are associated to at least one adjuvant.


As examples of adjuvants, a non limitative list is given below:

    • Aluminum-based compounds such as aluminum phosphate and aluminum hydroxide.
    • Immunostimulatory adjuvants like CpG, MPL and QS21 orMF59 which is a squalene oil-in water emulsions.
    • Freunds incomplete adjuvant (FIA) or Freund's complete adjuvant (FCA), can also be used.
    • Calcium phosphate in particular orthophosphates, metaphosphates or pyrophosphates and occasionally hydrogen or hydroxide ions.
    • Muramyl dipeptide (N-Acetyl muramyl-L-alanine-D-isoglutamine, MDP) and its synthetic analogs such as muramyl tripeptide phosphatidylethanolamine MTPPTdEtn),
    • Trehalose-6,6′-dimycolate,
    • Saponins (Triterpenoid glycosides) like QS-21
    • Stearyl Tyrosine (octadecyl ester hydrochloride salt of tyrosine)
    • Polysaccharides like:
      • Chitosan:
      • Inulin:
      • Beta-Glucans
      • Lipo-Polysaccharides or endotoxin
      • MGN-3Actinidia eriantha (AEPS):
      • Eldexomer:
      • CpG ODN
    • Liposomes like
      • Dehydration-rehydration liposome vesicles (DRVs)
      • Cytotoxic T lymphocyte (CTL)
      • CAF01
      • Liposomes containing lipid A (LA)
    • Lipid Polysine Core Peptides (LCP)
    • Cytokine like GM-CSF, IL-2, IL-12 or IL-15
    • Lipid A and Monophosphoryl Lipid A (MPL)
    • Lipopeptide which are molecules consisting of lipid connected to a peptide. (They are derived from the lipoprotein of bacterial cell wall.)
    • Exogenous Immunostimulatory Adjuvants which are compounds, or proteins that are not found in the human body like:
      • Proteosomes in particular Multiple Antigenic Peptides (MAP) or Exogenous Toxins.


Suitably, the total amount of mutated lentiviral ENV protein in a single dose of the immunogenic composition is 1-500 μg and/or the total amount of unfused polypeptides in a single dose of the immunogenic composition is 1-500 μg. In one embodiment, the total amount of all antigens in a single dose of the immunogenic composition is 1-1000 μg, 10-500 μg, 100-200 μg, or around 100 μg.


The amount of mutated feline lentiviral ENV protein in a dose of the immunogenic composition is selected as an amount which induces an immune response without significant, adverse side effects in typical recipients. Such amount will vary depending upon which specific immunogen is employed and the dosing or vaccination regimen that is selected. An optimal amount for a particular immunogenic composition can be ascertained by standard studies involving observation of relevant immune responses in infected animals.


Administration of the pharmaceutical composition can take the form of one or of more than one individual dose, for example as repeat doses of the same polypeptide containing composition, or in a heterologous “prime-boost” vaccination regime, including proteins and vectors. In one embodiment, the immunogenic composition of the invention is initially administered to a subject as two or three doses, wherein the doses are separated by a period of two weeks to three months, preferably one month. Conveniently, the composition is administered to an animal (for instance as a booster) every 6-24, or 9-18 months, for instance annually. For instance, the composition is administered to an animal (for instance as a booster) at six month or 1 year intervals. Suitably in this respect, subsequent administrations of the composition to the animal boost the immune response of earlier administrations of the composition to the same animal.


In an embodiment, the immunogenic composition of the invention is used as part of a prime-boost regimen for use in the treatment or prevention of disease or infection by FIV strains from one or more clades different from the one or more FIV clades in the immunogenic composition. Conveniently, the composition is the priming dose. Alternatively, the composition is the boosting dose.


Suitably, two or more priming and/or boosting doses are administered. A heterologous prime-boost regime uses administration of different forms of immunogenic composition or vaccine in the prime and the boost, each of which can itself include two or more administrations. The priming composition and the boosting composition will have at least one antigen in common, although it is not necessarily an identical form of the antigen, it can be a different form of the same antigen.


Prime boost immunisations according to the invention can be homologous prime-boost regimes or heterologous prime-boost regimes. Homologous prime-boost regimes utilize the same composition for prime and boost, for instance the immunogenic composition of the invention. Heterologous prime-boost regimes can be performed with a combination of protein and DNA-based formulations. Such a strategy is considered to be effective in inducing broad immune responses. Adjuvanted protein vaccines induce mainly antibodies and CD4+T cell immune responses, while delivery of DNA as a plasmid or a recombinant vector induces strong CD8+T cell responses.


Thus, the combination of protein and DNA vaccination can provide for a wide variety of immune responses. This is particularly relevant in the context of FIV, since neutralizing antibodies, CD4+T cells and CD8+T cells are thought to be important for the immune defense against FIV.


The invention also relates to a method for treating animals afflicted by pathologies related to lentiviral infections, comprising the administration to an animal in a need thereof of a pharmaceutically efficient amount of the pharmaceutical composition as defined above.


The invention also relates to a pharmaceutical composition as defined above, as a vaccine, for its use for the prevention or the traitement of FIV infection, or pathologies related to feline immunodeficiency.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance:


an isolated non naturally occurring mutated feline lentiviral ENV having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said mutated feline lentiviral ENV resulting from mutation of the transmembrane subunit (TM) of a wild type feline lentiviral ENV protein, the transmembrane subunit comprising an immunosuppressive domain (ISU) comprising the following amino acid sequence:









(SEQ ID NO: 343)









A-[I/M/T/L]-E-K-F-L-Y-T-A,







wherein the amino acids at the positions chosen among
    • the position 3 of SEQ ID NO: 343, and/or
    • the position 4 of SEQ ID NO: 343 and/or
    • the position 5 of SEQ ID NO: 343, and/or
    • the position 6 of SEQ ID NO: 343, and/or
    • the position 7 of SEQ ID NO: 343,


      are:
    • either deleted,
    • or substituted by another amino acid such that
      • the amino acid residue at position 3 of SEQ ID NO: 1 is substituted by A, F, G, L, R, C, D, H, I, K, M, N, P, Q, S, T, V, W or Y, and/or
      • the amino acid residue at position 4 of SEQ ID NO: 1 is substituted by A, F, G, L, R, C, D, E, H, I, M, N, P, Q, S, T, V, W or Y, and/or
      • the amino acid residue at position 5 of SEQ ID NO: 1 is substituted by A, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and/or
      • the amino acid residue at position 6 of SEQ ID NO: 1 is substituted by A, F, G, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and/or
      • the amino acid residue at position 7 of SEQ ID NO: 1 is substituted by A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V or W.


In another aspect, the invention relates to a method to obtain the active substance of a pharmaceutical composition, as defined above, consisting in modifying the immunosuppressive property of:


a wild-type feline lentiviral ENV protein,


or a fragment of said wild-type feline lentiviral ENV protein, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said ENV protein or fragment thereof presenting a transmembrane subunit (TM) comprising an immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 344)









A-[I/M/T/L]-XA-XB-XC-XD-XE-T-A,







wherein
    • XA is C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XB is C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XC is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XD is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XE is C, D, E, H, I, K, M, N, P, Q, 5, T, V, W or Y,


      said method comprising a step of introduction of at least one mutation of XA and/or XB and/or XC and/or XD and/or XE,


      to obtain:


an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


said mutated feline lentiviral ENV protein having at least 70% identity, preferably at least 80% identity, to one sequence SEQ ID NO: 5,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K or deleted, and/or
    • X3 is any amino acid different from F or deleted, and/or
    • X4 is any amino acid different from L or deleted, and/or
    • X5 is any amino acid different from Y or deleted.


In a particular embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, consisting in modifying the immunosuppressive property of a wild-type feline lentiviral ENV protein, or a fragment of said wild-type feline lentiviral ENV protein, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids, said ENV protein or fragment thereof presenting a transmembrane subunit (TM) comprising an immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 345)







[V/I]-[E/R]-A-[I/M/T/L]-XA-XB-XC-XD-XE-T-A-[F/L]-





A-M,







wherein
    • XA is C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XB is C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XC is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XD is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and
    • XE is C, D, E, H, I, K, M, N, P, Q, 5, T, V, W or Y,


said method comprising a step of introduction of at least one mutation of XA and/or XB and/or XC and/or XD and/or XE,


to obtain:


an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


said mutated feline lentiviral ENV protein having at least 70% identity, preferably at least 80% identity, to one sequence SEQ ID NO: 5,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 3)







[V/I]-[E/R]-A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A-[F/L]-





A-M,







wherein
    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K or deleted, and/or
    • X3 is any amino acid different from F or deleted, and/or
    • X4 is any amino acid different from L or deleted, and/or
    • X5 is any amino acid different from Y or deleted.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, consisting in modifying the immunosuppressive property of a wild-type feline lentiviral ENV protein, or a fragment of said wild-type feline lentiviral ENV protein, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said ENV protein or fragment thereof presenting a transmembrane subunit (TM) comprising an immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 346)









A-[I/M/T/L]-E-K-[F/P]-[L/V/I]-Y-T-A,






said method comprising a step of introduction of at least one mutation of E in position 3 and/or K in position 4 and/or [F/P] in position 5 and/or [L/V/I] in position 6 and/or Y in position 7,


to obtain:


an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K or deleted, and/or
    • X3 is any amino acid different from F or deleted, and/or
    • X4 is any amino acid different from L or deleted, and/or
    • X5 is any amino acid different from Y or deleted.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, consisting in modifying the immunosuppressive property of a wild-type feline lentiviral ENV protein, or a fragment of said wild-type feline lentiviral ENV protein, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids, said ENV protein or fragment thereof presenting a transmembrane subunit (TM) comprising an immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 347)







[V/I]-[E/R]-A-[I/M/T/L]-E-K-[F/P]-[L/V/I]-Y-T-A-





[F/L]-A-M,







said method comprising a step of introduction of at least one mutation of E in position 3 and/or K in position 4 and/or [F/P] in position 5 and/or [L/V/I] in position 6 and/or Y in position 7,


to obtain:


an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 3)







[V/I]-[E/R]-A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A-[F/L]-





A-M,







wherein
    • X1 is any amino acid different from E or deleted, and/or
    • X2 is any amino acid different from K or deleted, and/or
    • X3 is any amino acid different from F or deleted, and/or
    • X4 is any amino acid different from L or deleted, and/or
    • X5 is any amino acid different from Y or deleted.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X1 is any amino acid different from E or deleted, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is any amino acid different from F or deleted, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is any amino acid different from L or deleted, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is any amino acid different from Y or deleted, and X1, X2, X3 and X4 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, and X3, X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, and X2, X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X4 is any amino acid different from L or deleted, and X2, X3 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X5 is any amino acid different from Y or deleted, and X2, X3 and X4 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, and X1, X4 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, and X2, X4 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X5 is any amino acid different from Y or deleted, and X2, X3 and X4 are any amino acid, or
    • X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X2, X4 and X5 are any amino acid, or
    • X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X2, X3 and X4 are any amino acid, or
    • X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X2, X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, and X4 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, and X3 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X5 is any amino acid different from Y or deleted, and X3 and X4 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X2 and X5 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X2 and X4 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X2 and X3 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X1 and X5 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X1 and X4 are any amino acid, or
    • X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X1 and X3 are any amino acid, or
    • X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X1 and X2 are any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, and X5 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X5 is any amino acid different from Y or deleted, and X4 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X3 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted, and X2 is any amino acid, or
    • X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted, X5 is any amino acid different from Y or deleted and X1 is any amino acid, or
    • X1 is any amino acid different from E or deleted, X2 is any amino acid different from K or deleted, X3 is any amino acid different from F or deleted, X4 is any amino acid different from L or deleted and X5 is any amino acid different from Y or deleted.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids, said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X1 is R, G, L, A, F or deleted, and/or
    • X2 is R, G, L, A, F or deleted, and/or
    • X3 is R, G, L, A, or deleted, and/or
    • X4 is R, G, A, F or deleted, and/or
    • X5 is R, G, L, A, F or deleted.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 3)







[V/I]-[E/R]-A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A-[F/L]-





A-M,







wherein
    • X1 is R, G, L, A, F or deleted, and/or
    • X2 is R, G, L, A, F or deleted, and/or
    • X3 is R, G, L, A, or deleted, and/or
    • X4 is R, G, A, F or deleted, and/or
    • X5 is R, G, L, A, F or deleted.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X1 is R, G, L, A, F or deleted, and/or
    • X2 is G, L, F or deleted, and/or
    • X3 is R, G, L, A, or deleted, and/or
    • X4 is R, G, A, F or deleted, and/or
    • X5 is G, L, F or deleted.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X3 is R, G, L, A or deleted, X4 is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X4 is R, G, A, F or deleted, X3 is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X3 is R, G, L, A or deleted, X4 is R, G, A, F or deleted, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, 5, T, V, W or Y.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X3 is R, A or deleted, X4 is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X4 is R, A or deleted, X3 is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X3 is R, A or deleted, X4 is R, A or deleted, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X3 is R, X4 is L, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X4 is R, X3 is F, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, or
    • X3 is R, X4 is R, and X1, X2, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X3 is R, and X1 is E, X2 is K, X4 is L, X5 is Y, or
    • X4 is R, and X1 is E, X2 is K, X3 is F, X5 is Y, or
    • X3 is R, X4 is R, and X1 is E, X2 is K, X5 is Y.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein


X3 is R, and X1, X2, X4, X5 are A, F, G, L, R, C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above


wherein said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X3 is R, and X1 is E, X2 is K, X4 is L, X5 is Y.


In another embodiment, the invention concerns a method to obtain the active substance of a pharmaceutical composition, as defined above, consisting in modifying the immunosuppressive property of:


a wild-type feline lentiviral ENV protein,


or a fragment of said wild-type feline lentiviral ENV protein, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said ENV protein or fragment thereof presenting a transmembrane subunit (TM) comprising an immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 344)









A-[I/M/T/L]-XA-XB-XC-XD-XE-T-A,







wherein
    • XA is E, and
    • XB is K, and
    • XC is F or P, and
    • XD is L, V or I, and
    • XE is Y,


      said method comprising a step of introduction of at least one mutation of XA and/or XB and/or XC and/or XD and/or XE,


      to obtain:


an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity,


said mutated feline lentiviral ENV protein having at least 70% identity, preferably at least 80% identity, to one sequence SEQ ID NO: 5,


or a fragment of said isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,


said mutated ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:









(SEQ ID NO: 1)









A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,







wherein
    • X1 is any amino acid different from E, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is any amino acid different from K, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is any amino acid different from F, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is any amino acid different from L, and X1, X2, X3 and X5 are any amino acid, or
    • X5 is any amino acid different from Y, and X1, X2, X3 and X4 are any amino acid,


      in particular, wherein
    • X1 is R, G, L, A or F, and X2, X3, X4 and X5 are any amino acid, or
    • X2 is R, G, L, A or F, and X1, X3, X4 and X5 are any amino acid, or
    • X3 is R, G, L or A, and X1, X2, X4 and X5 are any amino acid, or
    • X4 is R, G, A or F, and X1, X2, X3 and X5 are any amino acid, or


      X5 is R, G, L, A or F and X1, X2, X3 and X4 are any amino acid.


In other aspect, the invention also relates to pharmaceutical compositions comprising as active substance a mutated lentiviral protein isolated from other animal lentiviruses, infecting bovine, equine, ovine or caprine animal species.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated bovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated bovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type bovine lentiviral ENV protein, said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 420,


said mutated bovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 421)









Y-L-ZA1-ZA2-ZA3-ZA4-ZA5-[I/V]-[R/H],







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from Y or F or deleted, and/or
    • ZA3 is any amino acid different from V or L or deleted, and/or
    • ZA4 is any amino acid different from E or A or deleted, and/or
    • ZA5 is any amino acid different from E or deleted,


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated bovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated bovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type bovine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: SEQ ID NO: 420,


said mutated bovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 422)









V-[S/T]-Y-L-ZA1-ZA2-ZA3-ZA4-ZA5-







[I/V]-[R/H]-[E/Q]-[K/L/V]-Q,







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from Y or F or deleted, and/or
    • ZA3 is any amino acid different from V or L or deleted, and/or
    • ZA4 is any amino acid different from E or A or deleted, and/or
    • ZA5 is any amino acid different from E or deleted,


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated bovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated bovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type bovine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 423,


said mutated bovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 424)









Y-L-ZA1-ZA2-ZA3-ZA4-ZA5-V-H,







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from F or deleted, and/or
    • ZA3 is any amino acid different from L or deleted, and/or
    • ZA4 is any amino acid different from A or deleted, and/or
    • ZA5 is any amino acid different from E or deleted,


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated bovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated bovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type bovine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 423,


said mutated bovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 425)









V-T-Y-L-ZA1-ZA2-ZA3-ZA4-ZA5-V-H-E-[L/V]-Q,







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from F or deleted, and/or
    • ZA3 is any amino acid different from L or deleted, and/or
    • ZA4 is any amino acid different from A or deleted, and/or
    • ZA5 is any amino acid different from E or deleted,


      in association with a pharmaceutically acceptable carrier.


In a particular embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated bovine lentiviral Env protein, or said fragment thereof, further comprises additional mutations of at least one amino acid chosen among the 5th, 6th, 7th, 8th and 9th amino acids, in particular the 7th amino acid, located upstream the amino acid ZA1 of SEQ ID NO: 421, SEQ ID NO: 422, SEQ ID NO: 424 or SEQ ID NO: 425.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance a nucleic acid molecule coding for a mutated bovine lentiviral ENV protein, or a fragment of said mutated bovine lentiviral ENV protein, as defined above.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated equine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated equine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type equine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 426,


said mutated equine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 427)









L-L-ZB1-ZB2-ZB3-ZB4-ZB5-[V/I]-E,







wherein,
    • ZB1 is any amino acid different from K or deleted, and/or
    • ZB2 is any amino acid different from E or deleted, and/or
    • ZB3 is any amino acid different from R or K or Q or deleted, and/or
    • ZB4 is any amino acid different from Q or deleted, and/or
    • ZB5 is any amino acid different from Q or L or K or deleted, and/or


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated equine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated equine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type equine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 426,


said mutated equine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 428)







V-[Q/R]-L-L-ZB1-ZB2-ZB3-ZB4-ZB5-[V/I]-E-E-T-F,







wherein,
    • ZB1 is any amino acid different from K or deleted, and/or
    • ZB2 is any amino acid different from E or deleted, and/or
    • ZB3 is any amino acid different from K or Q or deleted, and/or
    • ZB4 is any amino acid different from Q or deleted, and/or
    • ZB5 is any amino acid different from Q or L or K or deleted, and/or


      in association with a pharmaceutically acceptable carrier.


In a particular embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated equine lentiviral ENV protein, or said fragment thereof, further comprises additional mutations of at least one amino acid chosen among the 5th, 6th, 7th, 8th and 9th amino acids, in particular the 7th amino acid, located upstream the amino acid ZA1 of SEQ ID NO: 427 or SEQ ID NO: 428.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance a nucleic acid molecule coding for a mutated equine lentiviral ENV protein, or a fragment of said mutated equine lentiviral ENV protein, as defined above.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated caprine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated caprine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type caprine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 429,


said mutated caprine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 432)









R-[V/M-ZA1-ZA2-ZA3-ZA4-ZA5-R-[M/I],







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from A or T or V or deleted, and/or
    • ZA3 is any amino acid different from I or L or V or M or deleted, and/or
    • ZA4 is any amino acid different from T or V or M or I or deleted, and/or
    • ZA5 is any amino acid different from D or deleted,


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated caprine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated caprine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type caprine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 429,


said mutated caprine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 433)







V-A-R-[V/M]-ZA1-ZA2-ZA3-ZA4-ZA5-R-[M/I]-M-[L/I/V]-





Y,







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from A or T or V or deleted, and/or
    • ZA3 is any amino acid different from I or L or V or M or deleted, and/or
    • ZA4 is any amino acid different from T or V or M or I or deleted, and/or
    • ZA5 is any amino acid different from D or deleted,


      in association with a pharmaceutically acceptable carrier.


In a particular embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated caprine lentiviral ENV protein, or said fragment thereof, further comprises additional mutations of at least one amino acid chosen among the 5th, 6th, 7th, 8th and 9th amino acids, in particular the 7th amino acid, located upstream the amino acid ZA1 of SEQ ID NO: 432 or SEQ ID NO: 433.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance a nucleic acid molecule coding for a mutated caprine lentiviral ENV protein, or a fragment of said mutated caprine lentiviral ENV protein, as defined above.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated ovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated ovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type ovine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 430,


said mutated ovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 432)









R-[V/M]-ZA1-ZA2-ZA3-ZA4-ZA5-R-[M/I],







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from A or T or V or deleted, and/or
    • ZA3 is any amino acid different from I or L or V or M or deleted, and/or
    • ZA4 is any amino acid different from T or V or M or I or deleted, and/or
    • ZA5 is any amino acid different from D or deleted,


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated ovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated ovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type ovine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 430


said mutated ovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 433)







V-A-R-[V/M]-ZA1-ZA2-ZA3-ZA4-ZA5-R-[M/I]-M-[L/I/V]-





Y,







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from A or T or V or deleted, and/or
    • ZA3 is any amino acid different from I or L or V or M or deleted, and/or
    • ZA4 is any amino acid different from T or V or M or I or deleted, and/or
    • ZA5 is any amino acid different from D or deleted,


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated ovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated ovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type ovine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 431


said mutated ovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 432)









R-[V/M]-ZA1-ZA2-ZA3-ZA4-ZA5-R-[M/I],







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from A or T or V or deleted, and/or
    • ZA3 is any amino acid different from I or L or V or M or deleted, and/or
    • ZA4 is any amino acid different from T or V or M or I or deleted, and/or
    • ZA5 is any amino acid different from D or deleted,


      in association with a pharmaceutically acceptable carrier.


In another aspect, the invention also relates to a pharmaceutical composition comprising as active substance an isolated mutated ovine lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof,


said mutated ovine lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type ovine lentiviral ENV protein,


said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 431


said mutated ovine lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:









(SEQ ID NO: 433)







V-A-R-[V/M]-ZA1-ZA2-ZA3-ZA4-ZA5-R-[M/I]-M-[L/I/V]-





Y,







wherein,
    • ZA1 is any amino acid different from E or deleted, and/or
    • ZA2 is any amino acid different from A or T or V or deleted, and/or
    • ZA3 is any amino acid different from I or L or V or M or deleted, and/or
    • ZA4 is any amino acid different from T or V or M or I or deleted, and/or
    • ZA5 is any amino acid different from D or deleted,


      in association with a pharmaceutically acceptable carrier.


In a particular embodiment, the invention relates to a pharmaceutical composition as defined above, wherein said mutated ovine lentiviral ENV protein, or said fragment thereof, further comprises additional mutations of at least one amino acid chosen among the 5th, 6th, 7th, 8th and 9th amino acids, in particular the 7th amino acid, located upstream the amino acid ZA1 of SEQ ID NO: 432 or SEQ ID NO: 433.


In another aspect, the invention relates to a pharmaceutical composition comprising as active substance a nucleic acid molecule coding for a mutated ovine lentiviral ENV protein, or a fragment of said mutated ovine lentiviral ENV protein, as defined above.





LEGEND TO THE FIGURES


FIG. 1:


(A) FIV Envelope phylogeny, with the A-D subtypes indicated and the reference Petaluma strain boxed (from Pu et al, Journal of Feline Medicine and Surgery, 2005, 7:65-70).


(B) Structure of the FIV Envelope protein, delineation of the characteristic functional domains and alignment of the 64-aa immunosuppressive-containing domains from selected FIV Env proteins. The SU and TM subunits of the FIV Env are indicated, together with the fusion peptide, the transmembrane anchoring domain of the TM subunit, and the immunosuppressive domain (ISD).


The aligned sequences of the 64-aa immunosuppressive-containing domains correspond to fragments of the Envelope protein from 33 distinct strains (i.e. 33 different accession numbers), only 20 of these 64-aa fragments are different (i.e. 20 SEQ ID numbers have been given).



FIG. 2:


Immunosuppressive activity of the full-length FIV envelope protein (FIV Env) and of the 64 aa-long FIV envelope subdomain delineated in FIG. 1 (FIV64 Env). Immunosuppression was tested using the in vivo MCA205 tumor rejection assay (see scheme on top and Materials and Methods): MCA205 tumor cells are transduced with an expression vector (containing a selectable hygromycin gene) for the indicated Env protein or fragment, stably transduced cells are then selected for expression of the corresponding Env, and are finally engrafted into Balb/c mice (allogenic graft, normally resulting in tumor rejection); tumor growth/rejection is then monitored and provides an immunosuppressive index (calculated as indicated in Materials and Methods). Y-axis corresponds to the value of the immunosuppression index. Immunosuppression indexes are means of at least 3 independent experiments, with standard deviation. The Murine Leukemia Virus (MLV) envelope ectodomains, wild-type and mutant (Schlecht-Louf et al., Proc Natl Acad Sci USA. 2010, 107(8):3782-7) are used as an internal control.



FIG. 3:


Functional identification of the aminoacids in the FIV envelope 64 aa domain directly involved in immunosuppressive activity, and search for aminoacid substitutions inhibiting this activity. Immunosuppressive activity was tested as in FIG. 2, using the in vivo MCA205 tumor rejection assay (see Materials and Methods). The mutated aminoacids are indicated with their position (see sequence of FIV64 on top) and the nature of the substitution. Wwt and Wmut are controls (from the human endogenous retrovirus HERV-W Envelope, Mangeney et al., Proc Natl Acad Sci USA. 2007, 104(51):20534-9). Immunosuppression indexes correspond to 3 independent experiments, with standard deviation.



FIG. 4:


Antibody response of mice inoculated with cells expressing wild-type and mutant FIV64 Env domains (see scheme on top): MCA205 tumor cells are stably transduced as in FIG. 2 with expression vectors for the indicated FIV64 Env wt and mutants, and cells engrafted into C57B1/6 mice (syngenic graft, resulting in constant tumor growth without rejection); specific anti-FIV64 antibodies are quantitated from the collected blood samples as indicated in Materials and Methods. Results are representative of 2 independent experiments.



FIG. 5:


Antibody response of mice injected with MBP-FIV64, wt and mutants, recombinant proteins (see scheme on top): FIV64 Env, wt and mutants, fused with MBP are i.v. injected three times with a 1-week interval into Balb/C mice (10 μg per injection); mice are then blood-sampled and anti-MBP IgG levels are determined by ELISA using plates coated with MBP-LacZα (MBP-LacZα is a control MBP fusion with the 83aa LacZα fragment (α-subunit of E. coli β-galactosidase, see Materials and Methods). Results are representative of 2 independent experiments.



FIG. 6:


Expression of the FIV Env proteins, wt and mutants, at the surface of cells transfected with the indicated expression vectors (see scheme on top).


Expression profile of the wild-type and mutant FIV Env proteins was assayed by FACS analysis using full-length FIVenv-expressing vectors and an anti-SU FIVenv monoclonal antibody (Antibodies-online, BmbH, Germany). The data correspond to the Mean Fluorescence Intensity, from three independent experiments, with standard deviation.



FIG. 7:


Functional characterization of the FIV Env proteins, wild-type (wt) and mutants. Fusogenic activity of the FIVenv, wild-type and mutants, are measured by a cell-cell fusion assay and quantified by a fusion index (see scheme on top and Materials and Methods). The data correspond to three independent experiments, with standard deviation).





EXAMPLES
The FIV Envelope and a Delineated Subdomain of the TM Subunit are Immunosuppressive In Vivo

Identification of an immunosuppressive activity of the FIV envelope and of the domain within the FIV envelope responsible for this activity was achieved using an in vivo tumor rejection assay that we had previously used to demonstrate the immunosuppressive activity of the Env protein of oncoretroviruses (e.g. murine MoMLV and simian MPMV, see Mangeney and Heidmann, Proc Natl Acad Sci USA, 1998, 95:14920-14925; Mangeney et al., Proc Natl Acad Sci USA, 2007, 104(51):20534-9).


The rationale of the assay can be summarized as follows: while injection of MCA205 tumor cells (H-2b) into allogeneic Balb/c mice (H-2d) leads to the formation of no tumor or transient tumors that are rapidly rejected, injection of the same cells, but stably expressing an immunosuppressive retroviral Env protein, leads to the growth of larger tumors that persist for a longer time-in spite of the expression of the new exogenous antigen.


This difference is not associated with a difference in intrinsic cell growth rate since it is not observed in syngeneic C57BL/6 mice, and is immune system-dependent.


The extent of “immunosuppression” can be quantified by an index based on tumor size: (Aenv-Anone)/Anone, where Aenv and Anone are the mean areas at the peak of growth of tumors from Balb/c mice injected with env-expressing or control cells, respectively. A positive index indicates that env expression facilitates tumor growth, as a consequence of its immunosuppressive activity; a null or negative index points to no effect or even an inhibitory effect, respectively. The latter can be explained by a stimulation of the immune response of the host against the new foreign antigen, represented by a non-immunosuppressive Env protein, expressed at the surface of tumor cells. Accordingly, as illustrated in FIG. 2, it can be observed that the FIV env is immunosuppressive, and that this activity is carried by a 64-mer peptide of the transmembrane (TM) subunit of the envelope protein. The immunosuppression indexes are close to those found for non-lentiviral env proteins (such as that of the murine MLV retrovirus shown in the figure as a control, together with a non-immunosuppressive mutant).


Identification of the FIV Env Amino-Acids Critical for Immunosuppression and its Inhibition, Via R-Scanning

To characterize further the immunosuppressive domain (ISD) of the FIV env active in vivo, we analyzed the effect of a series of amino-acid substitutions within the FIV env 64 aa domain shown to carry the IS activity (see above). The 64-aa FIV env domain is embedded into the so-called ectodomain, which corresponds to the extracellular domain of the TM subunit, and consists in the α-helical domain involved in FIV TM trimerization, and the N-term part of the loop containing the 2 well-conserved, 6/7 aa-distant, cysteine residues found in most retroviral envelopes. Refine delineation of the amino acis responsible for IS activity was thus performed by arginine-scanning within this domain. As illustrated in FIG. 3, X-to-Arg substitutions resulted in a significant change in the IS activity of the FIVenv 64 aa domain, with maximum effect observed for the F687 position and to a lesser—but significant—extent for the adjacent E685, K686, L688 and Y689 positions. A series of other substitutions are performed at the F687 position, substitutions for the small A and G, and the hydrophobic L and F residues.


We also analyzed the effect of this series of amino-acid substitutions within the FIVenv 64 aa domain by measuring the production of anti-FIV env IgG antibodies in mice inoculated under syngenic conditions (C57B1/6 mice) with the cells expressing the wild-type and mutant FIVenv 64 aa domains. As illustrated in FIG. 4, X-to-Arg subsitutions resulted in an increase in the antibody response, consistent with an increased immunogenicity of FIVenv 64, with again a more important effect observed for the F687 position.


Additionally, a series of experiments were performed with recombinant proteins containing the FIVenv 64 domain, to assay the relative immunogenicity of the wt and mutants proteins. The domains were fused with the carrier MBP (Maltose Binding Protein), and the proteins were injected i.v. into Balb/C mice (see Materials and Methods). The immunosuppressive effects of the FIVenv 64 wt and mutants were assayed by measuring the inhibition of the anti-MBP antibody response raised in the injected mice. As illustrated in FIG. 5, a significant decrease—relative to the control (MBP-LacZα)—is observed with FIVenv 64 wt, consistent with its immunosuppressive activity acting in cis, whereas almost no reduction is observed for the F687R and L688R mutants, indicative of the loss of its IS activity. A similar effect is observed for the other mutants, although at a lesser extent.


Impact of the Identified Mutations on Env Protein Folding and Proper Expression as a Functional Transmembrane Protein

We then tested whether the above-mentioned substitutions alter the overall capacity of the FIV envelope to be expressed by an eucaryotic cell and to be exported at the cell membrane, by introducing the mutants into an expression vector for the full-length FIV envelope. A FACS analysis of cells transfected with the wild-type and the mutant FIV envelope genes inserted within a CMV-driven expression vector (see Materials and Methods) and using an anti-SU specific monoclonal antibody, demonstrated quantitative expression of the mutant envelopes at the cell surface (FIG. 6). In particular, this analysis indicated that the F687R and L688R mutations did not significantly altered the FIV Env structure and/or SU-TM interaction. Finally, the fusogenic activity of the FIV full-length envelopes was tested by a cell-cell fusion assay, in which cells prone to FIVenv-mediated fusion were tranfected as above with the corresponding expression vectors, and cell-cell fusion monitored 24-48 h post transfection by measuring the amount of multi-nucleated syncytia formed, with the process being quantified by a fusion index (see Materials and Methods). As illustrated in FIG. 7 and despite the conservation of the amount of env protein expressed at the cell surface (cf FIG. 6), variations in the fusion index exist among the various env mutants tested, although all of them remain fusion-positive,


Accordingly, the present investigation has clearly identified definite positions and definite substitutions within the FIV env resulting in the loss of its IS activity.


Being compatible with the conservation of the overall structure of the FIV Env protein, these substitutions should be introduced in all pharmaceutical preparations which include the Env protein as a vaccine antigen.


Materials and Methods

Mice and Cell Lines:


C57B1/6 and Balb/c mice, 6-10 weeks old, were obtained from Harlan (France). Mice were maintained in the animal facility of the Gustave Roussy Institute in accordance with institutional regulations. 293T (ATCC CRL11268), and MCA205 cells were cultured in DMEM supplemented with 10% fetal calf serum (Invitrogen), streptomycin (100 μg/m1) and penicillin (100 units/ml).


Plasmids Constructions:





    • phCMV-FIVenv: The pET34TF10 full feline immunodeficiency virus (FIV) genome clone (strain petaluma) (Talbott et al., Proc Natl Acad Sci USA, 1989, 86(15):5743-4747) served as template to generate full-length FIVenv PCR fragment using primers 1-2. This PCR fragment was digested with XhoI and MluI restriction enzymes to be ligated with phCMV vector opened with the same enzymes to generate a phCMV-FIVenv expression vector.

    • phCMV-FIVenv mutants: Mutated phCMV-FIVenv was obtained by successive





PCR using appropriate primers. A first series of PCRs was performed with phCMV-FIVenv as template with primer 3 and the reverse primer designed with the mutation (i.e. primers 5, 7, 9, 11, 13, 15, 17, 19, 21, 23); and primer 2 and the forward primer bearing the mutation (i.e. primers 4, 6, 8, 10, 12, 13, 14, 16, 18, 20, 22) to introduce the mutations E685A, E685R, K686A, K686R, F687A, F687R, L688A, L688R, Y689A and Y689R, respectively. The two PCR fragments bearing the same mutation were purified and mixed to be used as template for a subsequent PCR with primers 3 and 2. These PCR fragments were digested by SpeI and MluI restriction enzymes and purified. In the meantime the FIVenv fragment obtained after digestion of the phCMV-FIVenv with the XhoI and SpeI restriction enzymes was purified. A three-fragment ligation with phCMV opened with XhoI and MluI, the FIVenv fragment digested with XhoI and SpeI, and the SpeI-MluI-restricted PCR fragment with the desired mutation, was performed to obtain the phCMV-FIVenv mutants.

    • Mutations of the amino acids E685, K686 and Y689 into G, L or F, mutations of the amino acid F687 into G or L, and mutations of the amino acid L688 into G or F are similarly performed using appropriate primers pairs (i.e. primers 31-58).
    • pDFG-FIVenv and pDFG-FIVenv mutants: FIVenv and FIVenv mutant inserts were obtained by restriction with Agel and MluI of the phCMV-FIVenv and mutants. These inserts were ligated with pDFG-ectoSyncytin-1 (see above Mangeney et al, PNAS, 2007) opened with the same enzymes.
    • pDFG-FIV64 and pDFG-FIV112, wild-type and mutants: fragments of the FIVenv, wild-type and mutants, were PCR-amplified from phCMV-FIVenv using primers 24-25 or 24-26 to obtain FIV64 and FIV112 fragments, respectively. These PCR fragments were digested with SfiI and MluI and inserted into pDFG-ectoSyncytin-1 (see above Mangeney et al, PNAS, 2007) opened with the same enzymes.
    • pSIN-FIV64: This vector was based on the lentiviral vector pHR'SIN-cPPT-SEW described in Demaison et al. (Human Gene Therapy, 2002, 13:803-813). The pSIN-FIV64 vectors were obtained by insertion of the hygromycin resistance gene under the control of the phosphoglycerate kinase promoter (PGK) at the end of WPRE sequence of pHR'SIN-cPPT-SEW. Then the FIV-64 fragments, wt and mutants, were PCR-amplified from pDFG-FIV64 wt and mutants, using primers 27-28, digested with Bam HI and Not I, and inserted downstream the pSec sequence under control of the SFFV promoter.
    • pMal-MBP-FIV: Bacterial expression vectors for fusion proteins with E. coli maltose-binding protein (MBP) were constructed by ligation of a modified pMal-c2x (described in Center et al., Protein Sci, 1998, 7(7):1612-1619) opened with PstI and HindIll, with the FIV-env ectodomain, wt and mutants, obtained by PCR from phCMV Env wt and mutants, with primer pair 29-30. These pMal-MBP-FIV64 vectors encode a 64-residue long FIV-ectodomain fused to the C terminus of MBP through a trialanine linker. The empty pMal-c2x vector encodes the 85-residue long α-subunit of E. coli β-galactosidase fused to the C terminus of MBP and was used as a control (pMal-LacZa).


All the constructions were sequenced before use.









TABLE 2







Primer list










No
Name
Primer sequence (5′-3′)
SEQ ID NO:













1
FIV env Xho Age
atacatCTCGAGACCGGTccaactagaaccATGGCAGA
348



Kozak ATG (FOR)
AGGATTTGCAGCC





2
FIV env Stop Mlu
ATACATacgcgtTCATTCCTCCTCTTTTTCAGAC
349



(REV)
ATGCCAC





3
FIV-env-2000-FOR
TACTGCTATAGGGATGGTAACACAATACCAC
350




CAAG





4
FIV-env-E685A-
GATTAAAAGTAGAAGCTATGGCAAAATTTTT
351



FOR
GTATACAGC





5
FIV-env-E685A-
GCTGTATACAAAAATTTTGCCATAGCTTCTA
352



REV
CTTTTAATC





6
FIV-env-E685R-
GATTAAAAGTAGAAGCTATGAGAAAATTTTT
353



FOR
GTATACAGC





7
FIV-env-E685R-
GCTGTATACAAAAATTTTCTCATAGCTTCTAC
354



REV
TTTTAATC





8
FIV-env-K686A-
GTAGAAGCTATGGAAGCATTTTTGTATACAG
355



FOR
CTTTC





9
FIV-env-K686A-
GAAAGCTGTATACAAAAATGCTTCCATAGCT
356



REV
TCTAC





10
FIV-env-K686R-
GTAGAAGCTATGGAAAGATTTTTGTATACAG
357



FOR
CTTTC





11
FIV-env-K686R-
GAAAGCTGTATACAAAAATCTTTCCATAGCT
358



REV
TCTAC





12
FIV-env-F687A-
GAAGCTATGGAAAAAGCCTTGTATACAGCTT
359



FOR
TC





13
FIV-env-F687A-
GAAAGCTGTATACAAGGCTTTTTCCATAGCT
360



REV
TC





14
FIV-env-F687R-
GAAGCTATGGAAAAAAGATTGTATACAGCTT
361



FOR
TC





15
FIV-env-F687R-
GAAAGCTGTATACAATCTTTTTTCCATAGCTTC
362



REV





16
FIV-env-L688A-
GCTATGGAAAAATTTGCCTATACAGCTTTCG
363



FOR
CTATG





17
FIV-env-L688A-
CATAGCGAAAGCTGTATAGGCAAATTTTTCC
364



REV
ATAGC





18
FIV-env-L688R-
GCTATGGAAAAATTTAGGTATACAGCTTTCG
365



FOR
CTATG





19
FIV-env-L688R-
CATAGCGAAAGCTGTATACCTAAATTTTTCC
366



REV
ATAGC





20
FIV-env-Y689A-
CTATGGAAAAATTTTTGGCCACAGCTTTCGC
367



FOR
TATGC





21
FIV-env-Y689A-
GCATAGCGAAAGCTGTGGCCAAAAATTTTTC
368



REV
CATAG





22
FIV-env-Y689R-
CTATGGAAAAATTTTTGCGGACAGCTTTCGC
369



FOR
TATGC





23
FIV-env-Y689R-
GCATAGCGAAAGCTGTCCGCAAAAATTTTTC
370



REV
CATAG





24
FIV64-Sfi-FOR
GGTGACGCGGCCCAGCCGGCCgctatagaaaaggtga
371




ctggagcc





25
FIV64-Mlu-REV
ATACATACGCGTTTAccttgtccacaactcaagagg
372





26
FIVeq112-MLU-
ATACATACGCGTTTAccctgttttcccttgtacattattttg
373



REV





27
BamHI psec-FOR
ATA GGA TCC AGA ACC ATG GAG ACA GAC
435




ACA CTC





28
NotI FIV-REV
TAT GC GGCC GC TTA CCT TGT CCA CAA
436




CTC AAG





29
PstI FIV72-FOR
ATAGCTGCAGCCCAAGTTCTGGCAACCCAT
437





30
HindIII FIV72-REV
TATAAGCTTTTACCTTGTCCACAACTCAAG
438





31
FIV-env-G678A-
CATCAAGTACTAGTAATAGCATTAAAAGTAG
439



FOR
AAGCTATG





32
FIV-env-G678A-
CATAGCTTCTACTTTTAATGCTATTACTAGTA
440



REV
CTTGATG





33
FIV-env-E685G-
GATTAAAAGTAGAAGCTATGGGAAAATTTTT
441



FOR
GTATACAGC





34
FIV-env-E685G-
GCTGTATACAAAAATTTTCCCATAGCTTCTAC
442



REV
TTTTAATC





35
FIV-env-E685L-
GATTAAAAGTAGAAGCTATGTTAAAATTTTT
443



FOR
GTATACAGC





36
FIV-env-E685L-
GCTGTATACAAAAATTTTAACATAGCTTCTA
444



REV
CTTTTAATC





37
FIV-env-K686G-
GTAGAAGCTATGGAAGGATTTTTGTATACAG
445



FOR
CTTTC





38
FIV-env-K686G-
GAAAGCTGTATACAAAAATCCTTCCATAGCT
446



REV
TCTAC





39
FIV-env-K686L-
GTAGAAGCTATGGAATTATTTTTGTATACAG
447



FOR
CTTTC





40
FIV-env-K686L-
GAAAGCTGTATACAAAAATAATTCCATAGCT
448



REV
TCTAC





41
FIV-env-F687G-
GAAGCTATGGAAAAAGGATTGTATACAGCTT
449



FOR
TC





42
FIV-env-F687G-
GAAAGCTGTATACAATCCTTTTTCCATAGCTTC
450



REV





43
FIV-env-F687L-
GAAGCTATGGAAAAATTATTGTATACAGCTT
451



FOR
TC





44
FIV-env-F687L-
GAAAGCTGTATACAATAATTTTTCCATAGCTTC
452



REV





45
FIV-env-L688G-
GCTATGGAAAAATTTGGATATACAGCTTTCG
453



FOR
CTATG





46
FIV-env-L688G-
CATAGCGAAAGCTGTATATCCAAATTTTTCC
454



REV
ATAGC





47
FIV-env-Y689G-
CTATGGAAAAATTTTTGGGAACAGCTTTCGC
455



FOR
TATGC





48
FIV-env-Y689G-
GCATAGCGAAAGCTGTTCCCAAAAATTTTTC
456



REV
CATAG





49
FIV-env-Y689L-
CTATGGAAAAATTTTTGTTAACAGCTTTCGCT
457



FOR
ATGC





50
FIV-env-Y689L-
GCATAGCGAAAGCTGTTAACAAAAATTTTTC
458



REV
CATAG





51
FIV-env-E685F-
GATTAAAAGTAGAAGCTATGTTTAAATTTTT
459



FOR
GTATACAGC





52
FIV-env-E685F-
GCTGTATACAAAAATTTAAACATAGCTTCTA
460



REV
CTTTTAATC





53
FIV-env-K686F-
GTAGAAGCTATGGAATTTTTTTTGTATACAGC
461



FOR
TTTC





54
FIV-env-K686F-
GAAAGCTGTATACAAAAAAAATTCCATAGCT
462



REV
TCTAC





55
FIV-env-L688F-
GCTATGGAAAAATTTTTTTATACAGCTTTCGC
463



FOR
TATG





56
FIV-env-L688F-
CATAGCGAAAGCTGTATAAAAAAATTTTTCC
464



REV
ATAGC





57
FIV-env-Y689F-
CTATGGAAAAATTTTTGTTTACAGCTTTCGCT
465



FOR
ATGC





58
FIV-env-Y689F-
GCATAGCGAAAGCTGTAAACAAAAATTTTTC
466



REV
CATAG









Recombinant Proteins:


Recombinant proteins were produced using BL21 (DE3) Escherichia coli cells (Stratagene) and pMal-derived expression vectors (New England Biolabs, France). Recombinant WT and mutants TM subunit ectodomains were soluble and were purified on on cross-linked Amylose Resin (New England Biolabs, France) packed in column with PBS as a binding and washing buffer and 20 mM Tris-C1, 5 mM maltose, pH 7.5, as an elution buffer. Proteins were then dialysed against phosphate-buffered saline pH 7,4 (PBS), and endotoxins were removed using Endotrap Blue Resin (Hyglos GmbH, Germany) according to manufacturer's protocol.


Establishment of Env-Expressing Tumor Cells and MCA205 Tumor-Rejection Assay:


7.5×105 293 T cells were cotransfected with the env-expressing pDFG retroviral vector to be tested (1.75 μg) and expression vectors for the MLV proteins (0.55 μg for the amphotropic MLV env vector and 1.75 μg for the MLV gag and pol vector). 36 hours post-transfection, supernatants were harvested for infection of MCA205 tumor cells (2.5 ml of supernatant per 5×105 cells with 4 μg/m1 polybrene). Cells were maintained in selective medium (400 units/ml hygromycin) for 3 weeks, and then washed with PBS, trypsinized and inoculated subcutaneously in the shaved area of each mouse right flank as in Mangeney et al (see above PNAS 1998, PNAS 2007). Tumor growth was monitored by palpation twice or thrice weekly and tumor area (mm2) determined by measuring perpendicular tumor diameters. The extent of “immunosuppression” was quantified by an index based on tumor size: (Aenv-Anone)/Anone, where Aenv and Anone are the mean areas at the peak of growth of tumors from Balb/c mice injected with env-expressing or control cells, respectively.


Analysis of the Antibody Response of Mice Inoculated with MCA205-Tranduced Cells Expressing FIV64 Env Wild-Type and Mutants:


the MCA205-transduced cells were also inoculated as above into syngenic mice (C57B1/6) and sera were collected 1, 2, and 3 weeks after injection. The antibody response against FIV64 env was assayed by ELISA. Briefly, several dilutions of the sera were incubated 1 h at RT on a plate pre-coated with 1 ng/ml of MBP-FIV64, and antibody binding was analysed by using a labeled anti-mouse IgG secondary antibody (GE Healthcare, UK).


Analysis of FIVenv Expression:


3×105 293 T cells were transfected with 2 μg of the expression vector for the FIV envelope (phCMV) either wild-type or mutated at the indicated positions using Fugene HD (Roche). Cells were washed 16 h later and then harvested 2 days post-transfection using PBS-EDTA 5 mM. The SU1-30 monoclonal antibody (Antibodies online, BmbH, Germany) was used (1/200 dilution) to stain the FIV envelope. As a secondary antibody, we used the goat anti mouse IgG Alexa 488 (1/400) (Invitrogen). Fluorescence was acquired by flow cytometry using a FACS Calibur (BD Biosciences), and data analysed by the CellQuest software (BD Biosciences).


Cell-Cell Fusion Assays:


For cell-cell fusion assays, 5×104 to 1×105 cells seeded in 24-well plates were transfected by using lipofectamine LTX (Life technologies) with 250 ng of env expression plasmid. Fusion activity of each envelope protein was visualized 24 to 48 h after transfection by May-Grunwald and Giemsa staining, according to the manufacturer's instructions (Sigma). The fusion index, which represents the percentage of fusion events in a cell population, is defined as [(N−S)/T]×100), where N is the number of nuclei in the syncytia, S is the number of syncytia, and T is the total number of nuclei counted

Claims
  • 1. Pharmaceutical composition comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof, in particular said fragment of said isolated mutated feline lentiviral ENV protein comprising at least 40 amino acids, in particular at least 60 amino acids,said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:
  • 2. Pharmaceutical composition comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof, in particular said fragment of said isolated mutated feline lentiviral ENV protein comprising at least 40 amino acids, in particular at least 60 amino acids,said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:
  • 3. Pharmaceutical composition according to claim 1 comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof, said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:
  • 4. Pharmaceutical composition according to claim 1, wherein: X1 is A, F, G, L or R, X2 is A, F, G, L or R, and X3, X4 and X5 are any amino acid, orX1 is A, F, G, L or R, X3 is A, G, L or R, and X2, X4 and X5 are any amino acid, orX1 is A, F, G, L or R, X4 is A, F, G or R, and X2, X3 and X5 are any amino acid, orX1 is A, F, G, L or R, X5 is A, F, G, L or R, and X2, X3 and X4 are any amino acid, orX2 is A, F, G, L or R, X3 is A, G, L or R, and X1, X4 and X5 are any amino acid, orX2 is A, F, G, L or R, X4 is A, F, G or R, and X1, X3 and X5 are any amino acid, orX2 is A, F, G, L or R, X5 is A, F, G, L or R, and X1, X3 and X4 are any amino acid, orX3 is A, G, L or R, X4 is A, F, G or R, and X1, X2 and X5 are any amino acid, orX3 is A, G, L or R, X5 is A, F, G, L or R, and X1, X2 and X4 are any amino acid, orX4 is A, F, G or R, X5 is A, F, G, L or R, and X1, X2 and X3 are any amino acid.
  • 5. Pharmaceutical composition according to claim 1, wherein: X1 is A, G or R, and X2, X3, X4 and X5 are any amino acid, orX2 is A, G or R, and X1, X3, X4 and X5 are any amino acid, orX3 is A, G or R, and X1, X2, X4 and X5 are any amino acid, orX4 is A, G or R, and X1, X2, X3 and X5 are any amino acid, orX5 is A, G or R, and X1, X2, X3 and X4 are any amino acid,in particular:X1 is R, and X2, X3, X4 and X5 are any amino acid, orX2 is R, and X1, X3, X4 and X5 are any amino acid, orX3 is R, and X1, X2, X4 and X5 are any amino acid, orX4 is R, and X1, X2, X3 and X5 are any amino acid, orX5 is R, and X1, X2, X3 and X4 are any amino acid.
  • 6. Pharmaceutical composition according to claim 1, wherein: X1 is A, G or R, X2 is A, G or R, and X3, X4 and X5 are any amino acid, orX1 is A, G or R, X3 is A, G or R, and X2, X4 and X5 are any amino acid, orX1 is A, G or R, X4 is A, G or R, and X2, X3 and X5 are any amino acid, orX1 is A, G or R, X5 is A, G or R, and X2, X3 and X4 are any amino acid, orX2 is A, G or R, X3 is A, G or R, and X1, X4 and X5 are any amino acid, orX2 is A, G or R, X4 is A, G or R, and X1, X3 and X5 are any amino acid, orX2 is A, G or R, X5 is A, G or R, and X1, X3 and X4 are any amino acid, orX3 is A, G or R, X4 is A, G or R, and X1, X2 and X5 are any amino acid, orX3 is A, G or R, X5 is A, G or R, and X1, X2 and X4 are any amino acid, orX4 is A, G or R, X5 is A, G or R, and X1, X2 and X3 are any amino acid,in particular:X1 is R, X2 is R, and X3, X4 and X5 are any amino acid, orX1 is R, X3 is R, and X2, X4 and X5 are any amino acid, orX1 is R, X4 is R, and X2, X3 and X5 are any amino acid, orX1 is R, X5 is R, and X2, X3 and X4 are any amino acid, orX2 is R, X3 is R, and X1, X4 and X5 are any amino acid, orX2 is R, X4 is R, and X1, X3 and X5 are any amino acid, orX2 is R, X5 is R, and X1, X3 and X4 are any amino acid, orX3 is R, X4 is R, and X1, X2 and X5 are any amino acid, orX3 is R, X5 is R, and X1, X2 and X4 are any amino acid, orX4 is R, X5 is R, and X1, X2 and X3 are any amino acid.
  • 7. Pharmaceutical composition according to claim 1, wherein: X1 is A, G or R, and X2, X3, X4 and X5 are any amino acid different from A, G or R, orX2 is A, G or R, and X1, X3, X4 and X5 are any amino acid different from A, G or R, orX3 is A, G or R, and X1, X2, X4 and X5 are any amino acid different from A, G or R, orX4 is A, G or R, and X1, X2, X3 and X5 are any amino acid different from A, G or R, orX5 is A, G or R, and X1, X2, X3 and X4 are any amino acid different from A, G or R,in particular:X1 is R, and X2, X3, X4 and X5 are any amino acid different from A, G or R, orX2 is R, and X1, X3, X4 and X5 are any amino acid different from A, G or R, orX3 is R, and X1, X2, X4 and X5 are any amino acid different from A, G or R, orX4 is R, and X1, X2, X3 and X5 are any amino acid different from A, G or R, orX5 is R, and X1, X2, X3 and X4 are any amino acid different from A, G or R.
  • 8. Pharmaceutical composition according to claim 1, wherein: X1 is A, G or R, X2 is A, G or R, and X3, X4 and X5 are any amino acid different from A, G or R, orX1 is A, G or R, X3 is A, G or R, and X2, X4 and X5 are any amino acid different from A, G or R, orX1 is A, G or R, X4 is A, G or R, and X2, X3 and X5 are any amino acid different from A, G or R, orX1 is A, G or R, X5 is A, G or R, and X2, X3 and X4 are any amino acid different from A, G or R, orX2 is A, G or R, X3 is A, G or R, and X1, X4 and X5 are any amino acid different from A, G or R, orX2 is A, G or R, X4 is A, G or R, and X1, X3 and X5 are any amino acid different from A, G or R, orX2 is A, G or R, X5 is A, G or R, and X1, X3 and X4 are any amino acid different from A, G or R, orX3 is A, G or R, X4 is A, G or R, and X1, X2 and X5 are any amino acid different from A, G or R, orX3 is A, G or R, X5 is A, G or R, and X1, X2 and X4 are any amino acid different from A, G or R, orX4 is A, G or R, X5 is A, G or R, and X1, X2 and X3 are any amino acid different from A, G or R,in particular:X1 is R, X2 is R, and X3, X4 and X5 are any amino acid different from A, G or R, orX1 is R, X3 is R, and X2, X4 and X5 are any amino acid different from A, G or R, orX1 is R, X4 is R, and X2, X3 and X5 are any amino acid different from A, G or R, orX1 is R, X5 is R, and X2, X3 and X4 are any amino acid different from A, G or R, orX2 is R, X3 is R, and X1, X4 and X5 are any amino acid different from A, G or R, orX2 is R, X4 is R, and X1, X3 and X5 are any amino acid different from A, G or R, orX2 is R, X5 is R, and X1, X3 and X4 are any amino acid different from A, G or R, orX3 is R, X4 is R, and X1, X2 and X5 are any amino acid different from A, G or R, orX3 is R, X5 is R, and X1, X2 and X4 are any amino acid, different from A, G or R orX4 is R, X5 is R, and X1, X2 and X3 are any amino acid different from A, G or R.
  • 9. Pharmaceutical composition according to claim 1, wherein:
  • 10. Pharmaceutical composition according to claim 1, wherein:
  • 11. Pharmaceutical composition according to claim 1, wherein said isolated mutated feline lentiviral ENV protein or said fragment thereof, comprises one of the amino acid sequences SEQ ID NO: 28 to 171.
  • 12. Pharmaceutical composition according to claim 1, wherein said isolated mutated feline lentiviral ENV protein consists of one of the amino acid sequences: SEQ ID NO: 5 and SEQ ID NO: 317 to 342 and 374 to 419.
  • 13. Pharmaceutical composition comprising as active substance a nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, as defined in claim 1.
  • 14. Pharmaceutical composition according to claim 13, wherein said nucleic acid molecule is contained in a vector, said vector comprising means allowing the expression of the mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, as previously defined, in particular, wherein said vector is chosen among a canarypox vector, a pox vector, a fowlpox, an adenoviral vector, a lentiviral vector, a measles vector, a Sendaï virus vector, a Cytomegalovirus vector or a Modified Vaccinia Ankara vector.
  • 15. Pharmaceutical composition according to claim 14, comprising at least one nucleic acid molecule coding for a GAG protein and/or a PRO protein and/or a POL protein of a feline lentivirus, said lentivirus being preferably of the same origin as the mutated lentiviral ENV protein, in particular, whereinsaid nucleic acid molecule coding for a mutated feline lentiviral ENV protein, or a fragment of said mutated feline lentiviral ENV protein, is contained in the same vector as the one which also contains said at least one nucleic acid molecule coding for a GAG protein and/or a PRO protein and/or a POL protein.
  • 16. Pharmaceutical composition as defined in claim 1 for its use for stimulating an immune response in a host organism, in particular, for its use for the prevention or the treatment of lentiviral infection, preferably FIV infection,in particular, for its use as a vaccine, in particular against FIV infection.
  • 17. Method to obtain the active substance of a pharmaceutical composition, as defined in claim 1, consisting in modifying the immunosuppressive property of: a wild-type feline lentiviral ENV protein,or a fragment of said wild-type feline lentiviral ENV protein, said fragment comprising at least 40 amino acids, in particular at least 60 amino acids,said ENV protein or fragment thereof presenting a transmembrane subunit (TM) comprising an immunosuppressive domain (ISU) containing the following amino acid sequence:
  • 18. Pharmaceutical composition according to claim 2, comprising as active substance an isolated mutated feline lentiviral ENV protein having a decreased immunosuppressive activity, substantially no immunosuppressive activity or no immunosuppressive activity, or a fragment thereof, said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein,said mutated ENV protein having at least 70% identity, preferably at least 80% identity, to the sequence SEQ ID NO: 5,said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:
  • 19. Pharmaceutical composition according to claim 2, wherein: X1 is A, F, G, L or R, X2 is A, F, G, L or R, and X3, X4 and X5 are any amino acid, orX1 is A, F, G, L or R, X3 is A, G, L or R, and X2, X4 and X5 are any amino acid, orX1 is A, F, G, L or R, X4 is A, F, G or R, and X2, X3 and X5 are any amino acid, orX1 is A, F, G, L or R, X5 is A, F, G, L or R, and X2, X3 and X4 are any amino acid, orX2 is A, F, G, L or R, X3 is A, G, L or R, and X1, X4 and X5 are any amino acid, orX2 is A, F, G, L or R, X4 is A, F, G or R, and X1, X3 and X5 are any amino acid, orX2 is A, F, G, L or R, X5 is A, F, G, L or R, and X1, X3 and X4 are any amino acid, orX3 is A, G, L or R, X4 is A, F, G or R, and X1, X2 and X5 are any amino acid, orX3 is A, G, L or R, X5 is A, F, G, L or R, and X1, X2 and X4 are any amino acid, orX4 is A, F, G or R, X5 is A, F, G, L or R, and X1, X2 and X3 are any amino acid.
  • 20. Pharmaceutical composition according to claim 2, wherein: X1 is A, G or R, and X2, X3, X4 and X5 are any amino acid, orX2 is A, G or R, and X1, X3, X4 and X5 are any amino acid, orX3 is A, G or R, and X1, X2, X4 and X5 are any amino acid, orX4 is A, G or R, and X1, X2, X3 and X5 are any amino acid, orX5 is A, G or R, and X1, X2, X3 and X4 are any amino acid,in particular:X1 is R, and X2, X3, X4 and X5 are any amino acid, orX2 is R, and X1, X3, X4 and X5 are any amino acid, orX3 is R, and X1, X2, X4 and X5 are any amino acid, orX4 is R, and X1, X2, X3 and X5 are any amino acid, orX5 is R, and X1, X2, X3 and X4 are any amino acid.
Priority Claims (1)
Number Date Country Kind
13305767.9 Jun 2013 EP regional
PCT Information
Filing Document Filing Date Country Kind
PCT/EP2014/061924 6/6/2014 WO 00