Methods for producing antibodies which specifically bind to interleukin-9 receptor

Abstract

The invention describes nucleic acid sequences which code for the interleukin 9 receptor (IL9-R) molecule. These sequences may be used as probes to identify cells expressing the molecule, and as agents to transfect recipient cells.

Description

FIELD OF THE INVENTION

This invention relates to the reception of the cytokine known as interleukin 9 by cells, via its receptor. More particularly, it relates to the isolation of nucleic acid sequences which code for interleukin 9 receptor molecules ("IL-9R" hereafter). These sequences can be used, e.g. as a source for IL-9 receptor, and as probes for cells which respond to the cytokine. The complementary sequences can be used to inhibit expression as well as to probe for the coding sequences.

BACKGROUND AND PRIOR ART

The last decade has seen knowledge of the immune system and its regulation expand tremendously. One area of particular interest has been that of research on the proteins and glycoproteins which regulate the immune system. Perhaps the best known of these molecules, which are generically referred to as "growth factors", "cytokines", "leukotrines", "lymphokines", etc., is interleukin-2 ("IL-2"). See, e.g., U.S. Pat. No. 4,778,879 to Mertelsmann et al.; U.S. Pat. No. 4,490,289, to Stern; U.S. Pat. No. 4,518,584, to Mark et al.; and U.S. Pat. No. 4,851,512 to Miyaji et al. Additional patents have issued which relate to interleukin 1--("IL-1"), such as U.S. Pat. No. 4,808,611, to Cosman. The disclosure of all of these patents are incorporated by reference herein.

In order for molecules such as IL-2 and IL-1 to exert their effect on cells, it is now pretty much accepted that these must interact with molecules, located on cell membranes, referred to as receptors. Patents which exemplify disclosures of interleukin receptors include Honjo et al., U.S. Pat. No. 4,816,565; and Urdal et al., U.S. Pat. No. 4,578,335, the disclosures of which are incorporated by reference. Recently, Fanslow, et al., Science 248: 739-41 (May 11, 1990) presented data showing that the effect of IL-1 in vivo could be regulated via the administration of a soluble form of its receptor. The last paragraph of the Fanslow paper, the disclosure of which is incorporated by reference, describes the types of therapeutic efficacy administration of soluble IL-1 receptor ("IL-1R") is expected to have.

The lymphokine IL-9, previously referred to as "P40", is a T-cell derived molecule which was originally identified as a factor which sustained permanent antigen independent growth of T4 cell lines. See, e.g., Uyttenhove, et al., Proc. Natl. Acad. Sci. 85: 6934 (1988), and Van Snick et al., J. Exp. Med. 169: 363 (1989), the disclosures of which are incorporated by reference, as is that of Simpson et al., Eur. J. Biochem. 183: 715 (1989).

The activity of IL-9 was at first observed to act on restricted T4 cell lines, failing to show activity on CTLs or freshly isolated T cells. See, e.g., Uyttenhove et al., supra, and Schmitt et al., Eur. J. Immunol. 19: 2167 (1989). This range of activity was expanded when experiments showed that IL-9 and the molecule referred to as T cell growth Factor III ("TCGF III") are identical. IL-9 enhances the proliferative effect of bone marrow derived mast cells to "IL-3", as is described by Hultner et al., Eur. J. Immunol. and in U.S. patent application Ser. No. 498,182 filed Mar. 23, 1990 the disclosures of both being incorporated by reference herein. It was also found that the human form of IL-9 stimulates proliferation of megakaryoblastic leukemia. See Yang et al., Blood 74: 1880 (1989). Recent work on IL9 has shown that it also supports erythroid colony formation (Donahue et al., Blood 75(12): 2271-2275 (Jun. 15, 1990)); promotes the proliferation of myeloid erythroid burst formation (Williams et al., Blood 76: 306-311 (Sep. 1, 1990)); and supports clonal maturation of BFU.E's of adult and fetal origin (Holbrook et al., Blood 77(10): 2129-2134 (May 15, 1991)). Expression of IL9 has also been implicated in Hodgkin's disease and large cell anaplastic lymphoma (Merz et al., Blood 78(8): 1311-1317 (Sep. 1, 1990)).

The art teaches the cloning of receptors for various members of the interleukin family. Moseley et al. Cell 59: 335-348 (1989), teach the isolation of cDNA coding for IL-4 receptors, and analysis of both genomic DNA and RNA for these molecules. To do this, Moseley et al. worked with cells exhibiting up to 1 million receptor molecules per cell, and an N-terminal amino acid sequence for IL-4 receptor. Holmes et al., Science 253: 1278-1280 (1991), and Murphy et al., Science 253: 1280-1282 (1991) discuss cDNA for the IL-8 receptor. Murphy et al. proceeded via hybridization studies, using an oligonucleotide probe based upon rabbit IL-8R amino acid sequences to isolate the human counterpart. Holmes et al. used human neutrophil CDNA libraries followed by transfection in COS cells.

Gillis, "T-cell Derived Lymphokines" in Paul, ed., Fundamental Immunology, Second Edition (New York, 1989), at pages 632 et seq. gives an overview of interleukin receptors. This reference describes cDNA for the IL1 receptor, the IL2 receptor and the IL-6 receptor.

These studies indicate that several factors are important in attempting to identify and isolate a nucleic acid sequence coding for an interleukin receptor. Ideally, one has both the amino acid sequence for the receptor and a cell type with a high degree of expression of the receptor molecule.

In the case of the interleukin 9 receptor, while Druez et al., J. Immunol. 145: 2494-2499 (1990) have identified and characterized the receptor, an amino acid sequence of the molecule is not yet available. In addition, very few cell types are known which express IL9-R (Druez, su gra), and those that do, express it at very low levels. Thus, it is surprising that it now possible to identify and to isolate nucleic acid sequences which code for the interleukin 9 receptor. This is the key feature of the invention described herein, as will be seen from the disclosure which follows.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 presents Scatchard analysis of expression of murine IL9 receptor following transfection of COS cells.

FIG. 2 aligns deduced human SEQ ID No: 1D and murine IL-9R amino acid sequences.

FIGS. 3A and 3B compare the response of TS1 cells, both before and after transfection with DNA coding for human IL-9R.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

EXAMPLE 1

The murine T cell clone, TS1, described by, e.g., Uyttenhove et al., Proc. Natl. Acad. Sci. 85: 6934-6938 (1988) the disclosure of which is incorporated by reference, expresses approximately 200 high affinity binding sites for IL-9, i.e., it expresses the IL-9 receptor molecule. See Druez et al., J. Immunol. 145: 2494-2499 (1990). This cell line, while presenting few receptor molecules does show the highest density of IL9R of all cells tested, and thus was selected as a source of mRNA for constructing a cDNA library.

Poly(A)+ mRNA was extracted from TS1 cells, and was then converted to double stranded cDNA using random hexanucleotide primers, following Grubler et al, Gene 25: 263-269 (1983), the disclosure of which is incorporated by reference.

Following this, EcoRI adaptors were attached, and any cDNA larger than 1.5 kilobases was isolated by fractionation on a 5-20% potassium acetate gradient, following Aruffo et al., Proc. Natl. Acad. Sci. 84: 8573-8577 (1987).

The size selected cDNA was then inserted into the ECORI site of expression vector pCDSRa taught by Takebe et al., Mol. Cell Biol. 8: 466-472 (1988). This was then transfected into E. coli strain XL1-blue using standard transformation procedures. (Maniatis). In order to screen for clones expressing IL-9R, plasmid DNA from the cDNA library was tested for the ability to express IL-9 binding activity by expression in COS cells. Basically, the cDNA library was subfractionated into 100 pools of about 500 clones each, and the DNA was transfected using the DEAE-dextran-chloroquine method of Aruffo et al., supra, into 1.5.times.10.sup.5 COS cells, seeded on glass microscope slides. Cells were allowed to grow for 2-3 days, and were then tested for expression of IL-9R with .sup.125 I labelled, purified recombinant murine IL9. This labeled material was prepared following Bolton et al., Biochem. J. 133: 529-539 (1973). The cells were incubated for three hours at 20.degree. C. with 0.2 nM of this material, washed briefly, fixed, and then dipped into liquid photographic emulsion. The slides were exposed for 10 days, then developed and examined microscopically for autoradiographic grains.

This screening resulted in two positive pools out of 100. One positive pool showed a single positive cell, and the second one showed 33 positive cells. This latter pool was selected for further testing, and was divided, first into 100 pools of 15 clones each, after which a single positive pool was selected, and divided into 100 single clones.

EXAMPLE 2

Following the separating and replating described at the end of example 1, supra, the screening methodology described therein was employed on the replated cells, and led to identification of a clone containing a plasmid referred to as p9RA1. Since the "source" plasmid pCDSRa was known and characterized, it was possible using standard methodologies to identify the insert as 1900 base pairs in length.

EXAMPLE 3

Using the p9RA1 1900 base pair segment as a probe, additional screening was carried out to identify additional murine IL9R receptor cDNA clones. The methodology followed was that of Maniatis et al., Molecular Cloning, a Laboratory Manual (Cold Spring Harbor Laboratory, New York, 1982), where the p9RA1 probe was hybridized to two further cDNA libraries which were generated in the BstXI site of vector pCDM8 (Aruffo et al, supra), using oligo T or random primers, followed by high stringency washes.

This methodology resulted in the identification of six additional clones. Two of these were oligo-dT primed cDNAs, and are referred to as p9RB1, and p9RB3, and four random primed clones p9RC2, p9RC3, p9RC4 and p9RC9. The sizes of these clones are as follows:

p9RB1 1600 bp p9RB3 900 bp p9RC2 2000 bp p9RC3 1000 bp p9RC4 3000 bp p9RC9 2100 bp.

EXAMPLE 4

In order to make sure that clone p9RA1 and all subsequent clones did in fact express IL9R, Scatchard analysis was carried out on transfected COS cells, following Goodwin et al., Cell 60: 941-951 (1990). This analysis, shown in FIG. 1, identified a single class of binding sites with a Kd of 194 pM, when p9RA1 was used. This is slightly higher than the dissociation constant measured on TS1 cells previously, i.e., 67 pM.

When the largest cDNA was tested (i.e., the C4 clone), high affinity binding sites for IL9 were also identified, with a Kd of 126 pM.

EXAMPLE 5

Following the isolation of murine clones, tests were also carried out to isolate analogous human material. To do this, a megakaryoblast cell line, i.e., Mo7E was used as a source of mRNA to make double stranded cDNA as per example 1. The plasmid pRC/RSV was used to receive the cDNA. This cDNA library was screened, using p9RA1 as a probe, and hybridization was carried out using the same conditions described supra, except washes were carried out at low stringency (2.times.SSC, 0.1% SDS, 55.degree. C.). Six clones were isolated, i.e., ph9RA2, 3, 4, 5, 6 and 9, and sequenced. The clone ph9RA3 contained a 1566 base pair open reading frame, which showed 66% identity with murine p9RC4. The deduced murine and human protein sequences are shown in FIG. 2, with a 53% identity over 522 amino acids.

EXAMPLE 6

In order to test whether clone ph9RA3 actually did code for a human IL9 receptor, the clone was transfected into murine cell line TS1, using double pulse electroporation. In brief, 5.times.10.sup.6 TS1 cells were resuspended at 37.degree. C. in 0.8 ml of Dulbecco's modified Eagle's medium, supplemented with 10% fetal bovine serum, 50 mM 2-mercaptoethanol, 0.55 mM L-arginine, 0.24 mM L-asparagine, and 1.25 mM L-glutamine. Plasmid DNA (50 .mu.g) was added to the cells in 0.4 cm cuvettes just before electroporation. After a double electric pulse (750 V, 7452.OMEGA., 40 .mu.F and 100 V, 74.OMEGA., 2100 .mu.F), cells were immediately diluted in fresh medium supplemented with murine IL9. After 24 hours, cells were washed and cultured in the presence of G418, and mouse IL9. These conditions resulted in a frequency of transfection of approximately 1/10,000. Following selection with G418, transfected cells were maintained in human IL9, and a TS1 proliferation assay was performed using the methodology of Uyttenhove et al., Proc. Natl., Acad. Sci. USA 85: 6934-6938 (1988). If the CDNA expresses hIL9R, then cells should proliferate, while those which do not contain it should not.

FIGS. 3A and 3B show that original TS1 cells, unresponsive to 100 units/ml of human IL9, became responsive and proliferated after transfection with the human IL9R cDNA.

EXAMPLE 7

The sequence of clone p9RC4, presented as SEQ ID NO: 1, shows an open reading frame coding a 468 amino acid protein. The deduced amino acid sequence predicts two hydrophobic regions, one of which spans amino acids 15-40, and probably represents a signal peptide. The probability weight matrix of von Heyne, Nucl. Acids Res. 14: 4683-4690 (1986) predicts a cleavage site for the signal peptide between positions 37 and 39. The second hydrophobic domain spans amino acids 271-291. This is presumed to constitute the transmembrane domain.

The putative extracellular domain contains 233 amino acids, including 6 cysteine residues and two potential N-linked glycosylation sites at positions 116 and 155. A "WSEWS" amino acids 244-248 of SEQ ID No: 7 motif, i.e., "Trp-Ser-Glu-Trp-Ser" amino acids 244-248 of SEQ ID No: 7, typical of the hematopoietin receptor superfamily described by Idzerda et al., J. Exp. Med. 171: 861-873 (1990), is found at positions 244-248.

The cytoplasmic portion of the molecule is characterized by a high percentage of serine (13%), and proline (12.4%), as well as three potential protein kinase C phosphorylation sites at positions 294, 416 and 465.

Comparison of the various clones indicates that p9RA1 and p9RB3 contain an additional glutamine between position 192 and 193 as compared to p9RC4, but without a frameshift. This residue lies in the extracellular domain, but as example 4, supra shows, it does not appear to affect the affinity for ligand. There is a 22 nucleotide deletion at this position in p9RC2. These features, and a potential intronic sequence in p9RC9, suggest alternate splicing events.

The analysis of p9RB3 implies the existence of a soluble form of IL9R. The cDNA for this clone contains a large part of extracellular domain, but lacks nucleotides 651-1719, which code the end of the N-terminal domain, the transmembrane and the cytoplasmic domain.

Clone p9RA1 is different from all other clones in that there is a stop codon after alanine (378), which is followed by a 736 base pair sequence unrelated to any other cDNA's sequenced.

The sequences for the murine CDNA described in this example is provided as follows:

p9RC4 (SEQ ID NO: 1) p9RA1 (SEQ ID NO: 2) p9RB3 (SEQ ID NO: 3).

The deduced amino acid sequences encoded by these cDNAs are presented as SEQ ID NOS: 7, 8 and 9 respectively.

EXAMPLE 8

The CDNA for human IL9-R was also analyzed. As indicated supra, clone ph9RA3 showed 66% identity with murine p9RC4 and 53% homology on the amino acid sequence level. A putative cleavage site is positioned between amino acids 39 and 40. This site is conserved between species, as is the transmembrane domain, the two potential N-glycosylation sites, and the consensus sequence for the hematopoietic superfamily, all of which are described in Example 7.

The cytoplasmic portion of the protein seemed less conserved, and was much larger (231 amino acids) than the murine counterpart (177 residues). Due to a stretch of 9 consecutive serines in positions 431-439, there is a high percentage of serine in the molecule (11.2%).

Clones ph9RA2, 4, 6 and 9 confirmed the sequence derived from ph9RA3. The clone ph9RA5, however, has an 85 nucleotide deletion in positions 1063-1147, suggesting a truncated protein. The putative truncated protein would be 307 amino acids long, and contain the complete extracellular and transmembrane regions of IL9-R, 5 amino acids of the cytoplasmic domain, and 11 unrelated residues.

The clone referred to as pH9RA6 contains a short intervening sequence at the beginning of the DNA, which leads into a stop codon, in frame with the normal initiative codon. It also creates a new ATG triplet in frame with the downstream portion of the coding sequence. In the IL9R molecule, this yields a transcript with a unique N-terminal sequence, the rest of the sequence being identical to pH9RA3. Comparison of pH9RA6 and pH9RA3 shows that, after the initial methionine common to both clones, pH9RA6 contains an insert of 22 amino acids. These are followed by the sequence "GWTLESE . . ." amino acids 10-16 of SEQ ID No: 10 which is the sequence beginning at position 10 of pH9RA3.

The nucleic acid sequences for ph9RA3, ph9RA5 and pH9RA6 are presented as SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively. The deduced amino acid sequences encoded by these are presented as SEQ ID NOS: 10, 11 and 12.

The foregoing teaches the isolation of a nucleic acid sequence which codes for the interleukin-9 receptor. Both murine and the homology found therebetween (53%, with up to 67% in the extracellular region) suggests that nucleic acid sequences coding for IL9-R from other species could also be identified.

The following data deal with CDNA, but it will be seen that the sequences of the cDNA put one in position of mRNA, as the latter can be derived from the former based on well known rules regarding construction of the sequences. Given the cDNA information, it is presumed that one could also secure the genomic analog of the cDNAs.

The information provided herein also teaches construction of vectors, such as plasmids, which contain the nucleic acid sequences of interest, i.e., those coding for mammalian IL9R. Such vectors may contain components in addition to the coding sequence, such as promoters operably linked to the coding sequence, "markers", such as genes for antibiotic resistance or selection, including the thymidine kinase or "TK" gene, as well as others which will be known to the skilled artisan. The nucleic acid sequences and vectors may be used--as has been shown--to transfect various cell types, such as "COS", "CHO", Spodoptera fruaiperda or other insect cell lines. The sequences, either alone or in appropriate vectors, can be used to transfect a panoply of prokaryotic and eukaryotic cells.

The isolation of nucleic acid sequences coding for the IL-9 receptor makes it possible for investigators to carry out several lines of investigation which were not possible or much more difficult without these. For example, as pointed out supra, even on these cells which express it best, expression of IL-9R is low. Isolation of the gene makes it possible to transfect recipient cells, followed by overexpression, amplification, etc. This leads to sufficient expression on cell surfaces to permit immunization with these cells, and generation of an immunogenic response to IL-9R, including the production of antibodies. Isolation of the antibody producing cells, followed by standard techniques of hybridoma biology leads to production of IL-9R specific monoclonal antibodies.

The antibodies produced, be they polyclonal or monoclonal, can then be used in therapeutic methods to block IL-9 from binding to IL-9R molecules. As binding of IL-9 to cell surfaces is implicated in several pathological conditions, this is an important therapeutic goal.

In addition IL-9R specific antibodies can be used for both qualitative and quantitative measurement of IL-9R expression on cells, following known immunoassay protocols.

The examples supra show the existence of a soluble form of IL-9R. As with other soluble interleukin receptor molecules (see Fanslow et al., supra), this molecule can be used to prevent the binding of IL-9 to cell bound receptor, and thus interfere with the affect of IL-9 on a cell type, subpopulation, etc. As such, soluble IL-9R may be said to be an antagonist for IL-9.

Recent work has shown that the soluble form of one interleukin receptor, i.e., IL-6R, functions as an agonist. See Taga et al., Cell 58: 573-591 (Aug. 11, 1989). The soluble form of IL-9R might function in a similar manner. In addition the IL-9R molecule, either the soluble form or a solubilized form of the molecule may be used as an immunogen for generation of IL-9R specific antibodies. Either the entire receptor molecule, or an immunogenic portion thereof, can be used in an appropriate animal, such as a mouse, rabbit or guinea pig, to generate an immune response which includes antibody formation. The antibodies can then be purified using standard techniques. Alternatively, antibody producing B cells can be isolated and utilized in any of the standard methods for producing hybridomas, so as to lead to the generation of IL-9R specific monoclonal antibodies.

An assay is described suDra, in Example 6, in which IL-9R cDNA expression is assayed by measuring the responsiveness of a transfected cell line to IL9. This assay methodology provides a means for screening for various agonists and antagonists. In brief, a transfected cell sample containing a sequence coding for IL9R is contacted with a compound of interest. If the compound is an agonist, it will bind to the IL-9R molecule on the cell surface, and lead to the series of events usually associated with IL-9/IL-9R binding. To the same end, an antagonist can be assayed by combining the compound of interest with IL-9 and the cell sample to determine whether the IL-9 has diminished impact, or no impact. The assay for agonist/antagonist may be viewed as part of a broader invention wherein one may assay for molecules which compete for binding to IL-9R.

In addition to the coding sequences discussed herein, the invention also embraces sequences complementary to the coding sequences. These complements, which can be derived from the coding sequences themselves, may be used, e.g., as probes or as "anti-sense" inhibitors to prevent expression of the IL9R coding sequences. Other aspects of the invention will be clear to the skilled artisan, and do not require elaboration here.

The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, it being recognized that various modifications are possible within the scope of the invention.

__________________________________________________________________________# SEQUENCE LISTING- (1) GENERAL INFORMATION:- (iii) NUMBER OF SEQUENCES: 12- (2) INFORMATION FOR SEQ ID NO: 1:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 2281 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear#1: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- CTCCATGGCC CTGGGAAGAT GCATTGCGGA AGGTTGGACC TTGGAGAGAG TG - #GCGGTGAA 60- ACAGGTCTCC TGGTTCCTGA TCTACAGCTG GGTCTGCTCT GGAGTCTGCC GG - #GGAGTCTC 120- GGTCCCAGAG CAAGGAGGAG GAGGGCAGAA GGCTGGAGCA TTCACCTGTC TC - #AGCAACAG 180- TATTTACAGG ATCGACTGCC ACTGGTCGGC TCCAGAGCTG GGCCAGGAAT CC - #AGGGCCTG 240- GCTCCTCTTT ACCAGTAACC AGGTGACTGA AATCAAACAC AAATGCACCT TC - #TGGGACAG 300- TATGTGTACC CTGGTGCTGC CTAAAGAGGA GGTGTTCTTA CCTTTTGACA AC - #TTCACCAT 360- CACACTTCAC CGCTGCATCA TGGGACAGGA ACAGGTCAGC CTGGTGGACT CA - #CAGTACCT 420- GCCCAGGAGA CACATCAAGT TGGACCCACC CTCTGATCTG CAGAGCAATG TC - #AGCTCTGG 480- GCGTTGTGTC CTGACCTGGG GTATCAATCT TGCCCTGGAG CCATTGATCA CA - #TCCCTCAG 540- CTACGAGCTG GCCTTCAAGA GGCAGGAAGA GGCCTGGGAG GCCCGGCACA AG - #GACCGTAT 600- CGTTGGAGTG ACCTGGCTCA TCCTTGAAGC CGTCGAACTG AATCCTGGTT CC - #ATCTACGA 660- GGCCAGGCTG CGTGTCCAGA TGACTTTGGA GAGTTATGAG GACAAGACAG AG - #GGGGAATA 720- TTATAAGAGC CATTGGAGTG AGTGGAGCCA GCCCGTGTCC TTTCCTTCTC CC - #CAGAGGAG 780- ACAGGGCCTC CTGGTCCCAC GCTGGCAATG GTCAGCCAGC ATCCTTGTAG TT - #GTGCCCAT 840- CTTTCTTCTG CTGACTGGCT TTGTCCACCT TCTGTTCAAG CTGTCACCCA GG - #CTGAAGAG 900- AATCTTTTAC CAGAACATTC CATCTCCCGA GGCGTTCTTC CATCCTCTCT AC - #AGTGTGTA 960- CCATGGGGAC TTCCAGAGTT GGACAGGGGC CCGCAGAGCC GGACCACAAG CA - #AGACAGAA1020- TGGTGTCAGT ACTTCATCAG CAGGCTCAGA GTCCAGCATC TGGGAGGCCG TC - #GCCACACT1080- CACCTATAGC CCGGCATGCC CTGTGCAGTT TGCCTGCCTG AAGTGGGAGG CC - #ACAGCCCC1140- GGGCTTCCCA GGGCTCCCAG GCTCAGAGCA TGTGCTGCCG GCAGGGTGTC TG - #GAGTTGGA1200- AGGACAGCCA TCTGCCTACC TGCCCCAGGA GGACTGGGCC CCACTGGGCT CT - #GCCAGGCC1260- CCCTCCTCCA GACTCAGACA GCGGCAGCAG CGACTATTGC ATGTTGGACT GC - #TGTGAGGA1320- ATGCCACCTC TCAGCCTTCC CAGGACACAC CGAGAGTCCT GAGCTCACGC TA - #GCTCAGCC1380- TGTGGCCCTT CCTGTGTCCA GCAGGGCCTG ACACCTACCA AGGGATGTGG GC - #ATTCTCTT1440- CCCTCCTATC CTCGGATGGC ACCAGACACA GTCTCTGCGT GTCTCTGCTA GG - #TGCACCAT1500- GTCTGTTTTG GGGAGATGAA CGAAAGGCCC CAGGCTGACC CTGGGGTGCG TG - #TGGAACTC1560- CGGAGAGGAG GCAGCTGTGC ACGGATCAGA GGCAATGCGG ATGGAAGCAG TA - #GACTGTGC1620- CTTACCCCCC TGCTCTGCCT TTGTGGTGGG GATGCCTCCA GGGTCAGCAT CT - #TAACATCG1680- CCTTCGCTTC TCTTGTCTTT CTGGCTCTGT CCCAGGCCTG AAAAAAGAAT GT - #GACAAGCA1740- GCCTGGTCTG TTCTTCCACC CCTAAAGGGC TGGCCTGGGC CCAGGGACAC TG - #ATGAGACA1800- ACATTGGTGA AGTGTCCCTT TTCAGTGCCT TTCCCATTAA GACCAGAAGG GA - #CGCTTTTG1860- ACTGCAGGCT GTGGGTGGCT GGGTACGGAG GGAATGATGG AGCTTTGAGC AG - #GTGGGGTT1920- GTCCATCTTT GAGCTTTTGG GGTTCCAAGA TCAGCTGGAA GGAGTCTCAC CG - #ACTGATTC1980- AAAGAAGTCT TACCCATCTG TGATATTTTC TTTCCTGGTG CCGTGATAAA AC - #ACCGTGAC2040- CAAAAATGAC TTACAAAAGG AAGAGTTGGC TTGGTTTAAG GTTCCAGAGG TG - #TGGAGACA2100- TGGCAGCCAG CGGCACACAT GGCAGTGAGG ACAGGAAGCT GAGAGCTCAC AT - #CTCAACCA2160- AAAGTTGAGT GAACTGAAAG TACTATCCCC TCCCCCACCC CAACTCCAGC AA - #GGCTCCAC2220- CCCCCTGAAG GTTCCATGCC TCCCTAAACA GCTCGGCCAA ATAGAGACCA AG - #TGTTCAAA2280# 2281- (2) INFORMATION FOR SEQ ID NO: 2:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1905 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear#2: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- CACCTCCTGG CTGGGGCTGC CTGAGACTCT CCATGGCCCT GGGAAGATGC AT - #TGCGGAAG 60- GTTGGACCTT GGAGAGAGTG GCGGTGAAAC AGGTCTCCTG GTTCCTGATC TA - #CAGCTGGG 120- TCTGCTCTGG AGTCTGCCGG GGAGTCTCGG TCCCAGAGCA AGGAGGAGGA GG - #GCAGAAGG 180- CTGGAGCATT CACCTGTCTC AGCAACAGTA TTTACAGGAT CGACTGCCAC TG - #GTCGGCTC 240- CAGAGCTGGG CCAGGAATCC AGGGCCTGGC TCCTCTTTAC CAGTAACCAG GT - #GACTGAAA 300- TCAAACACAA ATGCACCTTC TGGGACAGTA TGTGTACCCT GGTGCTGCCT AA - #AGAGGAGG 360- TGTTCTTACC TTTTGACAAC TTCACCATCA CACTTCACCG CTGCATCATG GG - #ACAGGAAC 420- AGGTCAGCCT GGTGGACTCA CAGTACCTGC CCAGGAGACA CATCAAGTTG GA - #CCCACCCT 480- CTGATCTGCA GAGCAATGTC AGCTCTGGGC GTTGTGTCCT GACCTGGGGT AT - #CAATCTTG 540- CCCTGGAGCC ATTGATCACA TCCCTCAGCT ACGAGCTGGC CTTCAAGAGG CA - #GGAAGAGG 600- CCTGGGAGCA GGCCCGGCAC AAGGACCGTA TCGTTGGAGT GACCTGGCTC AT - #CCTTGAAG 660- CCGTCGAACT GAATCCTGGT TCCATCTACG AGGCCAGGCT GCGTGTCCAG AT - #GACTTTGG 720- AGAGTTATGA GGACAAGACA GAGGGGGAAT ATTATAAGAG CCATTGGAGT GA - #GTGGAGCC 780- AGCCCGTGTC CTTTCCTTCT CCCCAGAGGA GACAGGGCCT CCTGGTCCCA CG - #CTGGCAAT 840- GGTCAGCCAG CATCCTTGTA GTTGTGCCCA TCTTTCTTCT GCTGACTGGC TT - #TGTCCACC 900- TTCTGTTCAA GCTGTCACCC AGGCTGAAGA GAATCTTTTA CCAGAACATT CC - #ATCTCCCG 960- AGGCGTTCTT CCATCCTCTC TACAGTGTGT ACCATGGGGA CTTCCAGAGT TG - #GACAGGGG1020- CCCGCAGAGC CGGACCACAA GCAAGACAGA ATGGTGTCAG TACTTCATCA GC - #AGGCTCAG1080- AGTCCAGCAT CTGGGAGGCC GTCGCCACAC TCACCTATAG CCCGGCATGC CC - #TGTGCAGT1140- TTGCCTGCCT GAAGTGGGAG GCCACAGCGT GAGAAGGGAC AGCCAGCCAC TC - #AGTGCGTG1200- GGCTTAGATT GGGAAGAGAC CTCCCAAGCA GCTTCCCCTC CTCCCCAGCC CC - #TGCCATTC1260- ACCCCTGCTG GCCGTCCATC CCCAGGATCC ACTGTGGAGC CAAGCCCACA GA - #CCCGGCCT1320- GATTCAGCTC TGACACTCGC TGCGCTGCTC CGTTGTGAAC TTTGGCCAAG TC - #ACCACTTT1380- TACCTCAGCT TCCTCCTGTG AGAACAGGGT TGCCTTAGAG TTGCCTAATC CC - #TAAGGAGA1440- CTGAGACAAA CTTGTCTGCA AATATCTATC CGATGTATAT TGTTAGGAGC TC - #GAGGGTCC1500- GTGGGTGGGC GGGGCAGGGG GGTGGGGATG CGGTTGGCGC ATATCACTGT GT - #CAACAGCC1560- AGAGCCTTCC TCCATGTCTC AACCAACACT CTCCAAGCTG AATTCTCAGG CT - #GAACTCAC1620- TGTCACCTGT GAAGTAAACC CCGGCAGACC TGGAAGATTG GTGGTAGGAT TG - #TGGAGGTT1680- GCAGGGAGCA TGCTCAGTGG GCACTAGTTG CCTGCTGGGT ACCAGGAGAT GC - #TTGTGCCC1740- TGAGGTATCT TTAAGAACTA TCACGGAATT GGACTGGGAG CTCAGGAGAG AG - #CTTGGTAG1800- ACTGGCAGTG TCAGTGAAAC AGTTATTTAG CCAAGAACAA CATTCCTGGG GC - #TGGGGACA1860# 1905CC TGGAACATGG GAGGTTGTAA GTTCG- (2) INFORMATION FOR SEQ ID NO: 3:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1214 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear#3: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- ATGGCCCTGG GAAGATGCAT TGCGGAAGGT TGGACCTTGG AGAGAGTGGC GG - #TGAAACAG 60- GTCTCCTGGT TCCTGATCTA CAGCTGGGTC TGCTCTGGAG TCTGCCGGGG AG - #TCTCGGTC 120- CCAGAGCAAG GAGGAGGAGG GCAGAAGGCT GGAGCATTCA CCTGTCTCAG CA - #ACAGTATT 180- TACAGGATCG ACTGCCACTG GTCGGCTCCA GAGCTGGGCC AGGAATCCAG GG - #CCTGGCTC 240- CTCTTTACCA GTAACCAGGT GACTGAAATC AAACACAAAT GCACCTTCTG GG - #ACAGTATG 300- TGTACCCTGG TGCTGCCTAA AGAGGAGGTG TTCTTACCTT TTGACAACTT CA - #CCATCACA 360- CTTCACCGCT GCATCATGGG ACAGGAACAG GTCAGCCTGG TGGACTCACA GT - #ACCTGCCC 420- AGGAGACACA TCAAGTTGGA CCCACCCTCT GATCTGCAGA GCAATGTCAG CT - #CTGGGCGT 480- TGTGTCCTGA CCTGGGGTAT CAATCTTGCC CTGGAGCCAT TGATCACATC CC - #TCAGCTAC 540- GAGCTGGCCT TCAAGAGGCA GGAAGAGGCC TGGGAGCAGG CCCGGCACAA GG - #ACCGTATC 600- GTTGGAGTGA CCTGGCTCAT CCTTGAAGCC GTCGAACTGA ATCCTGAAAA AA - #GAATGTGA 660- CAAGCAGCCT GGTCTGTTCT TCCACCCCTA AAGGGCTGGC CTGGGCCCAG GG - #ACACTGAT 720- GAGACAACAT TGGTGAAGTG TCCCTTTTCA GTGCCTTTCC CATTAAGACC AG - #AAGGGACG 780- CTTTTGACTG CAGGCTGTGG GTGGCTGGGT ACGGAGGGAA TGATGGAGCT TT - #GAGCAGGT 840- GGGGTTGTCC ATCTTTGAGC TTTTGGGTTC CAAGATCAGC TGGAAGGAGT CT - #CACCGACT 900- GATTCAAAGA AGTCTTACCC ATCTGTGATA TTTTCTTTCC TGGTGCCGTG AT - #AAAACACC 960- GTGACCAAAA ATGACTTACA AAAGGAAGAG TTGGCTTGGT TTAAGGTTCC AG - #AGGTGTGG1020- AGACATGGCA GCCAGCGGCA CACATGGCAG TGAGGACAGG AAGCTGAGAG CT - #CACATCTC1080- AACCAAAAGT TGAGTGAACT GAAAGTACTA TCCCCTCCCC CACCCCAACT CC - #AGCAAGGC1140- TCCACCCCCC TGAAGGTTCC ATGCCTCCCT AAACAGCTCG GCCAAATAGA GA - #CCAAGTGT1200# 1214- (2) INFORMATION FOR SEQ ID NO: 4:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1947 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear#4: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- AGCAGCTCTG TAATGCGCTT GTGGTTTCAG ATGTGGGCGG CCTGTGTGAA CC - #TGTCGTGC 60- AAAGCTCACG TCACCAACTG CTGCAGTTAT CTCCTGAATC AGGCTGAGGG TC - #TTTGCTGT 120- GCACCCAGAG ATAGTTGGGT GACAAATCAC CTCCAGGTTG GGGATGCCTC AG - #ACTTGTGA 180- TGGGACTGGG CAGATGCATC TGGGAAGGCT GGACCTTGGA GAGTGAGGCC CT - #GAGGCGAG 240- ACATGGGCAC CTGGCTCCTG GCCTGCATCT GCATCTGCAC CTGTGTCTGC TT - #GGGAGTCT 300- CTGTCACAGG GGAAGGACAA GGGCCAAGGT CTAGAACCTT CACCTGCCTC AC - #CAACAACA 360- TTCTCAGGAT CGATTGCCAC TGGTCTGCCC CAGAGCTGGG ACAGGGCTCC AG - #CCCCTGGC 420- TCCTCTTCAC CAGCAACCAG GCTCCTGGCG GCACACATAA GTGCATCTTG CG - #GGGCAGTG 480- AGTGCACCGT CGTGCTGCCA CCTGAGGCAG TGCTCGTGCC ATCTGACAAT TT - #CACCATCA 540- CTTTCCACCA CTGCATGTCT GGGAGGGAGC AGGTCAGCCT GGTGGACCCG GA - #GTACCTGC 600- CCCGGAGACA CGTTAAGCTG GACCCGCCCT CTGACTTGCA GAGCAACATC AG - #TTCTGGCC 660- ACTGCATCCT GACCTGGAGC ATCAGTCCTG CCTTGGAGCC AATGACCACA CT - #TCTCAGCT 720- ATGAGCTGGC CTTCAAGAAG CAGGAAGAGG CCTGGGAGCA GGCCCAGCAC AG - #GGATCACA 780- TTGTCGGGGT GACCTGGCTT ATACTTGAAG CCTTTGAGCT GGACCCTGGC TT - #TATCCATG 840- AGGCCAGGCT GCGTGTCCAG ATGGCCACAC TGGAGGATGA TGTGGTAGAG GA - #GGAGCGTT 900- ATACAGGCCA GTGGAGTGAG TGGAGCCAGC CTGTGTGCTT CCAGGCTCCC CA - #GAGACAAG 960- GCCCTCTGAT CCCACCCTGG GGGTGGCCAG GCAACACCCT TGTTGCTGTG TC - #CATCTTTC1020- TCCTGCTGAC TGGCCCGACC TACCTCCTGT TCAAGCTGTC GCCCAGGGTG AA - #GAGAATCT1080- TCTACCAGAA CGTGCCCTCT CCAGCGATGT TCTTCCAGCC CCTCTACAGT GT - #ACACAATG1140- GGAACTTCCA GACTTGGATG GGGGCCCACA GGGCCGGTGT GCTGTTGAGC CA - #GGACTGTG1200- CTGGCACCCC ACAGGGAGCC TTGGAGCCCT GCGTCCAGGA GGCCACTGCA CT - #GCTCACTT1260- GTGGCCCAGC GCGTCCTTGG AAATCTGTGG CCCTGGAGGA GGAACAGGAG GG - #CCCTGGGA1320- CCAGGCTCCC GGGGAACCTG AGCTCAGAGG ATGTGCTGCC AGCAGGGTGT AC - #GGAGTGGA1380- GGGTACAGAC GCTTGCCTAT CTGCCACAGG AGGACTGGGC CCCCACGTCC CT - #GACTAGGC1440- CGGCTCCCCC AGACTCAGAG GGCAGCAGGA GCAGCAGCAG CAGCAGCAGC AG - #CAGCAACA1500- ACAACAACTA CTGTGCCTTG GGCTGCTATG GGGGATGGCA CCTCTCAGCC CT - #CCCAGGAA1560- ACACACAGAG CTCTGGGCCC ATCCCAGCCC TGGCCTGTGG CCTTTCTTGT GA - #CCATCAGG1620- GCCTGGAGAC CCAGCAAGGA GTTGCCTGGG TGCTGGCTGG TCACTGCCAG AG - #GCCTGGGC1680- TGCATGAGGA CCTCCAGGGC ATGTTGCTCC CTTCTGTCCT CAGCAAGGCT CG - #GTCCTGGA1740- CATTCTAGGT CCCTGACTCG CCAGATGCAT CATGTCCATT TTGGGAAAAT GG - #ACTGAAGT1800- TTCTGGAGCC CTTGTCTGAG ACTGAACCTC CTGAGAAGGG GCCCCTAGCA GC - #GGTCAGAG1860- GTCCTGTCTG GATGGAGGCT GGAGGCTCCC CCCTCAACCC CTCTGCTCAG TG - #CCTGTGGG1920# 1947 CAGC ATCCTGG- (2) INFORMATION FOR SEQ ID NO: 5:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1683 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear#5: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- ATGGGACTGG GCAGATGCAT CTGGGAAGGC TGGACCTTGG AGAGTGAGGC CC - #TGAGGCGA 60- GACATGGGCA CCTGGCTCCT GGCCTGCATC TGCATCTGCA CCTGTGTCTG CT - #TGGGAGTC 120- TCTGTCACAG GGGAAGGACA AGGGCCAAGG TCTAGAACCT TCACCTGCCT CA - #CCAACAAC 180- ATTCTCAGGA TCGATTGCCA CTGGTCTGCC CCAGAGCTGG GACAGGGCTC CA - #GCCCCTGG 240- CTCCTCTTCA CCAGCAACCA GGCTCCTGGC GGCACACATA AGTGCATCTT GC - #GGGGCAGT 300- GAGTGCACCG TCGTGCTGCC ACCTGAGGCA GTGCTCGTGC CATCTGACAA TT - #TCACCATC 360- ACTTTCCACC ACTGCATGTC TGGGAGGGAG CAGGTCAGCC TGGTGGACCC GG - #AGTACCTG 420- CCCCGGAGAC ACGTTAAGCT GGACCCGCCC TCTGACTTGC AGAGCAACAT CA - #GTTCTGGC 480- CACTGCATCC TGACCTGGAG CATCAGTCCT GCCTTGGAGC CAATGACCAC AC - #TTCTCAGC 540- TATGAGCTGG CCTTCAAGAA GCAGGAAGAG GCCTGGGAGC AGGCCCAGCA CA - #GGGATCAC 600- ATTGTCGGGG TGACCTGGCT TATACTTGAA GCCTTTGAGC TGGACCCTGG CT - #TTATCCAT 660- GAGGCCAGGC TGCGTGTCCA GATGGCCACA CTGGAGGATG ATGTGGTAGA GG - #AGGAGCGT 720- TATACAGGCC AGTGGAGTGA GTGGAGCCAG CCTGTGTGCT TCCAGGCTCC CC - #AGAGACAA 780- GGCCCTCTGA TCCCACCCTG GGGGTGGCCA GGCAACACCC TTGTTGCTGT GT - #CCATCTTT 840- CTCCTGCTGA CTGGCCCGAC CTACCTCCTG TTCAAGCTGT CGCCCAGACT TG - #GATGGGGG 900- CCCACAGGGC CGGTGTGCTG TTGAGCCAGG ACTGTGCTGG CACCCCACAG GG - #AGCCTTGG 960- AGCCCTGCGT CCAGGAGGCC ACTGCACTGC TCACTTGTGG CCCAGCGCGT CC - #TTGGAAAT1020- CTGTGGCCCT GGAGGAGGAA CAGGAGGGCC CTGGGACCAG GCTCCCGGGG AA - #CCTGAGCT1080- CAGAGGATGT GCTGCCAGCA GGGTGTACGG AGTGGAGGGT ACAGACGCTT GC - #CTATCTGC1140- CACAGGAGGA CTGGGCCCCC ACGTCCCTGA CTAGGCCGGC TCCCCCAGAC TC - #AGAGGGCA1200- GCAGGAGCAG CAGCAGCAGC AGCAGCAGCA GCAACAACAA CAACTACTGT GC - #CTTGGGCT1260- GCTATGGGGG ATGGCACCTC TCAGCCCTCC CAGGAAACAC ACAGAGCTCT GG - #GCCCATCC1320- CAGCCCTGGC CTGTGGCCTT TCTTGTGACC ATCAGGGCCT GGAGACCCAG CA - #AGGAGTTG1380- CCTGGGTGCT GGCTGGTCAC TGCCAGAGGC CTGGGCTGCA TGAGGACCTC CA - #GGGCATGT1440- TGCTCCCTTC TGTCCTCAGC AAGGCTCGGT CCTGGACATT CTAGGTCCCT GA - #CTCGCCAG1500- ATGCATCATG TCCATTTTGG GAAAATGGAC TGAAGTTTCT GGAGCCCTTG TC - #TGAGACTG1560- AACCTCCTGA GAAGGGGCCC CTAGCAGCGG TCAGAGGTCC TGTCTGGATG GA - #GGCTGGAG1620- GCTCCCCCCT CAACCCCTCT GCTCAGTGCC TGTGGGGAGC AGCCTCTACC CT - #CAGCATCC1680# 1683- (2) INFORMATION FOR SEQ ID NO: 6:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1997 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear#6: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- CCAGGTTGGG GATGCCTCAG ACTTGTGATG GGACTGGGCA GATGCATCTG GG - #AAGGTCCT 60- GGTGGTGACT CCAACCCTGC CCTCACATAT CCCAAGAGCA GCTGACTGCG CT - #TCCCCATT 120- CCCACCTTTC CAGTAACTGC TGCAAGAACG GACAGACACT GCTGCAGAGA AC - #TTGCCACG 180- GTGTTTCATG CTGTGGCTGG TGGTTCCAGG CTGCACGCTC CATTCTAGGA AA - #GGGGCCCT 240- CAGCCAGTCC CTTGCAGGCT GGACCTTGGA GAGTGAGGCC CTGAGGCGAG AC - #ATGGGCAC 300- CTGGCTCCTG GCCTGCATCT GCATCTGCAC CTGTGTCTGC TTGGGAGTCT CT - #GTCACAGG 360- GGAAGGACAA GGGCCAAGGT CTAGAACCTT CACCTGCCTC ACCAACAACA TT - #CTCAGGAT 420- CGATTGCCAC TGGTCTGCCC CAGAGCTGGG ACAGGGCTCC AGCCCCTGGC TC - #CTCTTCAC 480- CAGCAACCAG GCTCCTGGCG GCACACATAA GTGCATCTTG CGGGGCAGTG AG - #TGCACCGT 540- CGTGCTGCCA CCTGAGGCAG TGCTCGTGCC ATCTGACAAT TTCACCATCA CT - #TTCCACCA 600- CTGCATGTCT GGGAGGGAGC AGGTCAGCCT GGTGGACCCG GAGTACCTGC CC - #CGGAGACA 660- CGTTAAGCTG GACCCGCCCT CTGACTTGCA GAGCAACATC AGTTCTGGCC AC - #TGCATCCT 720- GACCTGGAGC ATCAGTCCTG CCTTGGAGCC AATGACCACA CTTCTCAGCT AT - #GAGCTGGC 780- CTTCAAGAAG CAGGAAGAGG CCTGGGAGCA GGCCCAGCAC AGGGATCACA TT - #GTCGGGGT 840- GACCTGGCTT ATACTTGAAG CCTTTGAGCT GGACCCTGGC TTTATCCATG AG - #GCCAGGCT 900- GCGTGTCCAG ATGGCCACAC TGGAGGATGA TGTGGTAGAG GAGGAGCGTT AT - #ACAGGCCA 960- GTGGAGTGAG TGGAGCCAGC CTGTGTGCTT CCAGGCTCCC CAGAGACAAG GC - #CCTCTGAT1020- CCCACCCTGG GGGTGGCCAG GCAACACCCT TGTTGCTGTG TCCATCTTTC TC - #CTGCTGAC1080- TGGCCCGACC TACCTCCTGT TCAAGCTGTC GCCCAGGGTG AAGAGAATCT TC - #TACCAGAA1140- CGTGCCCTCT CCAGCGATGT TCTTCCAGCC CCTCTACAGT GTACACAATG GG - #AACTTCCA1200- GACTTGGATG GGGGCCCACA GGGCCGGTGT GCTGTTGAGC CAGGACTGTG CT - #GGCACCCC1260- ACAGGGAGCC TTGGAGCCCT GCGTCCAGGA GGCCACTGCA CTGCTCACTT GT - #GGCCCAGC1320- GCGTCCTTGG AAATCTGTGG CCCTGGAGGA GGAACAGGAG GGCCCTGGGA CC - #AGGCTCCC1380- GGGGAACCTG AGCTCAGAGG ATGTGCTGCC AGCAGGGTGT ACGGAGTGGA GG - #GTACAGAC1440- GCTTGCCTAT CTGCCACAGG AGGACTGGGC CCCCACGTCC CTGACTAGGC CG - #GCTCCCCC1500- AGACTCAGAG GGCAGCAGGA GCAGCAGCAG CAGCAGCAGC AGCAGCAACA AC - #AACAACTA1560- CTGTGCCTTG GGCTGCTATG GGGGATGGCA CCTCTCAGCC CTCCCAGGAA AC - #ACACAGAG1620- CTCTGGGCCC ATCCCAGCCC TGGCCTGTGG CCTTTCTTGT GACCATCAGG GC - #CTGGAGAC1680- CCAGCAAGGA GTTGCCTGGG TGCTGGCTGG TCACTGCCAG AGGCCTGGGC TG - #CATGAGGA1740- CCTCCAGGGC ATGTTGCTCC CTTCTGTCCT CAGCAAGGCT CGGTCCTGGA CA - #TTCTAGGT1800- CCCTGACTCG CCAGATGCAT CATGTCCATT TTGGGAAAAT GGACTGAAGT TT - #CTGGAGCC1860- CTTGTCTGAG ACTGAACCTC CTGAGAAGGG GCCCCTAGCA GCGGTCAGAG GT - #CCTGTCTG1920- GATGGAGGCT GGAGGCTCCC CCCTCAACCC CTCTGCTCAG TGCCTGTGGG GA - #GCAGCCTC1980# 1997 G- (2) INFORMATION FOR SEQ ID NO: 7:- (i) SEQUENCE CHARACTERISTICS:#acid residuesLENGTH: 468 amino (B) TYPE: amino acid (D) TOPOLOGY: linear#7: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- Met Ala Leu Gly Arg Cys Ile Ala Glu Gly Tr - #p Thr Leu Glu Arg Val#15- Ala Val Lys Gln Val Ser Trp Phe Leu Ile Ty - #r Ser Trp Val Cys Ser# 30- Gly Val Cys Arg Gly Val Ser Val Pro Glu Gl - #n Gly Gly Gly Gly Gln# 45- Lys Ala Gly Ala Phe Thr Cys Leu Ser Asn Se - #r Ile Tyr Arg Ile Asp# 60- Cys His Trp Ser Ala Pro Glu Leu Gly Gln Gl - #u Ser Arg Ala Trp Leu# 80- Leu Phe Thr Ser Asn Gln Val Thr Glu Ile Ly - #s His Lys Cys Thr Phe# 95- Trp Asp Ser Met Cys Thr Leu Val Leu Pro Ph - #e Glu Glu Val Phe Leu# 110- Pro Phe Asp Asn Phe Thr Ile Thr Leu His Ar - #g Cys Ile Met Gly Gln# 125- Glu Gln Val Ser Leu Val Asp Ser Gln Tyr Le - #u Pro Arg Arg His Ile# 140- Lys Leu Asp Pro Pro Ser Asp Leu Gln Ser As - #n Val Ser Ser Gly Arg145 1 - #50 1 - #55 1 -#60- Cys Val Leu Thr Trp Gly Ile Asn Leu Ala Le - #u Glu Pro Leu Ile Thr# 175- Ser Leu Ser Tyr Glu Leu Ala Phe Lys Arg Gl - #n Glu Glu Ala Trp Glu# 190- Ala Arg His Lys Asp Arg Ile Val Gly Val Th - #r Trp Leu Ile Leu Glu# 205- Ala Val Glu Leu Asn Pro Gly Ser Ile Tyr Gl - #u Ala Arg Leu Arg Val# 220- Gln Met Thr Leu Glu Ser Tyr Glu Asp Lys Th - #r Glu Gly Glu Tyr Tyr225 2 - #30 2 - #35 2 -#40- Lys Ser His Trp Ser Glu Trp Ser Gln Pro Va - #l Ser Phe Pro Ser Pro# 255- Gln Arg Arg Gln Gly Leu Leu Val Pro Arg Tr - #p Gln Trp Ser Ala Ser# 270- Ile Leu Val Val Val Pro Ile Phe Leu leu Le - #u Thr Gly Phe Val His# 285- Leu Leu Phe Lys Leu Ser Pro Arg Leu Lys Ar - #g Ile Phe Tyr Gln Asn# 300- Ile Pro Ser Pro Glu Ala Phe Phe His Pro Le - #u Tyr Ser Val Tyr His305 3 - #10 3 - #15 3 -#20- Gly Asp Phe Gln Ser Trp Thr Gly Ala Arg Ar - #g Ala Gly Pro Gln Ala# 335- Arg Gln Asn Gly Val Ser Thr Ser Ser Ala Gl - #y Ser Glu Ser Ser Ile# 350- Trp Glu Ala Val Ala Thr Leu Thr Tyr Ser Pr - #o Ala Cys Pro Val Gln# 3650- Phe Ala Cys Leu Lys Trp Glu Ala Thr Ala Pr - #o Gly Phe Pro Gly Leu# 380- Pro Gly Ser Glu His Val Leu Pro Ala Gly Cy - #s Leu Glu Leu Glu Gly385 3 - #90 3 - #95 4 -#00- Gln Pro Ser Ala Tyr Leu Pro Gln Glu Asp Tr - #p Ala Pro Leu Gly Ser# 415- Ala Arg Pro Pro Pro Pro Asp Ser Asp Ser Gl - #y Ser Ser Asp Tyr Cys# 430- Met Leu Asp Cys Cys Glu Glu Cys His Leu Se - #r Ala Phe Pro Gly His# 445- Thr Glu Ser Pro Glu Leu Thr Leu Ala Gln Pr - #o Val Ala Leu Pro Val# 460- Ser Ser Arg Ala465- (2) INFORMATION FOR SEQ ID NO: 8:- (i) SEQUENCE CHARACTERISTICS:#acid residuesLENGTH: 379 amino (B) TYPE: amino acid (D) TOPOLOGY: linear#8: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- Met Ala Leu Gly Arg Cys Ile Ala Glu Gly Tr - #p Thr Leu Glu Arg Val#15- Ala Val Lys Gln Val Ser Trp Phe Leu Ile Ty - #r Ser Trp Val Cys Ser# 30- Gly Val Cys Arg Gly Val Ser Val Pro Glu Gl - #n Gly Gly Gly Gly Gln# 45- Lys Ala Gly Ala Phe Thr Cys Leu Ser Asn Se - #r Ile Tyr Arg Ile Asp# 60- Cys His Trp Ser Ala Pro Glu Leu Gly Gln Gl - #u Ser Arg Ala Trp Leu# 80- Leu Phe Thr Ser Asn Gln Val Thr Glu Ile Ly - #s His Lys Cys Thr Phe# 95- Trp Asp Ser Met Cys Thr Leu Val Leu Pro Ph - #e Glu Glu Val Phe Leu# 110- Pro Phe Asp Asn Phe Thr Ile Thr Leu His Ar - #g Cys Ile Met Gly Gln# 125- Glu Gln Val Ser Leu Val Asp Ser Gln Tyr Le - #u Pro Arg Arg His Ile# 140- Lys Leu Asp Pro Pro Ser Asp Leu Gln Ser As - #n Val Ser Ser Gly Arg145 1 - #50 1 - #55 1 -#60- Cys Val Leu Thr Trp Gly Ile Asn Leu Ala Le - #u Glu Pro Leu Ile Thr# 175- Ser Leu Ser Tyr Glu Leu Ala Phe Lys Arg Gl - #n Glu Glu Ala Trp Glu# 190- Gln Ala Arg His Lys Asp Arg Ile Val Gly Va - #l Thr Trp Leu Ile Leu# 205- Glu Ala Val Glu Leu Asn Pro Gly Ser Ile Ty - #r Glu Ala Arg Leu Arg# 220- Val Gln Met Thr Leu Glu Ser Tyr Glu Asp Ly - #s Thr Glu Gly Glu Tyr225 2 - #30 2 - #35 2 -#40- Tyr Lys Ser His Trp Ser Glu Trp Ser Gln Pr - #o Val Ser Phe Pro Ser# 255- Pro Gln Arg Arg Gln Gly Leu Leu Val Pro Ar - #g Trp Gln Trp Ser Ala# 270- Ser Ile Leu Val Val Val Pro Ile Phe Leu Le - #u Leu Thr Gly Phe Val# 285- His Leu Leu Phe Lys Leu Ser Pro Arg Leu Ly - #s Arg Ile Phe Tyr Gln# 300- Asn Ile Pro Ser Pro Glu Ala Phe Phe His Pr - #o Leu Tyr Ser Val Tyr305 3 - #10 3 - #15 3 -#20- His Gly Asp Phe Gln Ser Trp Thr Gly Ala Ar - #g Arg Ala Gly Pro Gln# 335- Ala Arg Gln Asn Gly Val Ser Thr Ser Ser Al - #a Gly Ser Glu Ser Ser# 350- Ile Trp Glu Ala Val Ala Thr Leu Thr Tyr Se - #r Pro Ala Cys Pro Val# 365- Gln Phe Ala Cys Leu Lys Trp Glu Ala Thr Al - #a# 375- (2) INFORMATION FOR SEQ ID NO: 9:- (i) SEQUENCE CHARACTERISTICS:#acid residues) LENGTH: 219 amino (B) TYPE: amino acid (D) TOPOLOGY: linear#9: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- Met Ala Leu Gly Arg Cys Ile Ala Glu Gly Tr - #p Thr Leu Glu Arg Val#15- Ala Val Lys Gln Val Ser Trp Phe Leu Ile Ty - #r Ser Trp Val Cys Ser# 30- Gly Val Cys Arg Gly Val Ser Val Pro Glu Gl - #n Gly Gly Gly Gly Gln# 45- Lys Ala Gly Ala Phe Thr Cys Leu Ser Asn Se - #r Ile Tyr Arg Ile Asp# 50- Cys His Trp Ser Ala Pro Glu Leu Gly Gln Gl - #u Ser Arg Ala Trp Leu# 80- Leu Phe Thr Ser Asn Gln Val Thr Glu Ile Ly - #s His Lys Cys Thr Phe# 95- Trp Asp Ser Met Cys Thr Leu Val Leu Pro Ph - #e Glu Glu Val Phe Leu# 110- Pro Phe Asp Asn Phe Thr Ile Thr Leu His Ar - #g Cys Ile Met Gly Gln# 125- Glu Gln Val Ser Leu Val Asp Ser Gln Tyr Le - #u Pro Arg Arg His Ile# 140- Lys Leu Asp Pro Pro Ser Asp Leu Gln Ser As - #n Val Ser Ser Gly Arg145 1 - #50 1 - #55 1 -#60- Cys Val Leu Thr Trp Gly Ile Asn Leu Ala Le - #u Glu Pro Leu Ile Thr# 175- Ser Leu Ser Tyr Glu Leu Ala Phe Lys Arg Gl - #n Glu Glu Ala Trp Glu# 190- Ala Arg His Lys Lys Asp Arg Ile Val Gly Va - #l Thr Trp Leu Ile Leu# 205- Glu Ala Val Glu Leu Asn Pro Glu Lys Arg Me - #t# 215- (2) INFORMATION FOR SEQ ID NO: 10:- (i) SEQUENCE CHARACTERISTICS:#acid residuesLENGTH: 522 amino (B) TYPE: amino acid (D) TOPOLOGY: linear- (xi) SEQUENCE DESCRIPTION: SEQ ID NO: - # 10:- Met Gly Leu Gly Arg Cys Ile Trp Glu Gly Tr - #p Thr Leu Glu Ser Glu#15- Ala Leu Arg Arg Asp Met Gly Thr Trp Leu Le - #u Ala Cys Ile Cys Ile# 30- Cys Thr Cys Val Cys Leu Gly Val Ser Val Th - #r Gly Glu Gly Gln Gly# 45- Pro Arg Ser Arg Thr Phe Thr Cys Leu Thr As - #n Asn Ile Leu Arg Ile# 60- Asp Cys His Trp Ser Ala Pro Glu Leu Gly Gl - #n Gly Ser Ser Pro Trp# 80- Leu Leu Phe Thr Ser Asn Gln Ala Pro Gly Gl - #y Thr His Lys Cys Ile# 95- Leu Arg Gly Ser Glu Cys Thr Val Val Leu Pr - #o Pro Glu Ala Val Leu# 110- Val Pro Ser Asp Asn Phe Thr Ile Thr Phe Hi - #s His Cys Met Ser Gly# 125- Arg Glu Gln Val Ser Leu Val Asp Pro Glu Ty - #r Leu Pro Arg Arg His# 140- Val Lys Leu Asp Pro Pro Ser Asp Leu Gln Se - #r Asn Ile Ser Ser Gly145 1 - #50 1 - #55 1 -#60- His Cys Ile Leu Thr Trp Ser Ile Ser Pro Al - #a Leu Glu Pro Met Thr# 175- Thr Leu Leu Ser Tyr Glu Leu Ala Phe Lys Ly - #s Gln Glu Glu Ala Trp# 190- Glu Gln Ala Gln His Arg Asp His Ile Val Gl - #y Val Thr Trp Leu Ile# 205- Leu Glu Ala Phe Glu Val Asp Pro Gly Phe Il - #e His Glu Ala Arg Leu# 220- Arg Val Gln Met Ala Thr Leu Glu Asp Asp Va - #l Val Glu Glu Glu Arg225 2 - #30 2 - #35 2 -#40- Tyr Thr Gly Gln Trp Ser Glu Trp Ser Gln Pr - #o Val Cys Phe Gln Ala# 255- Pro Gln Arg Gln Gly Pro Leu Ile Pro Pro Tr - #p Gly Trp Pro Gly Asn# 270- Thr Leu Val Ala Val Ser Ile Phe Leu Leu Le - #u Thr Gly Pro Thr Tyr# 285- Leu Leu Phe Lys Leu Ser Pro Arg Val Lys Ar - #g Ile Phe Tyr Gln Asn# 300- Val Pro Ser Pro Ala Met Phe Phe Gln Pro Le - #u Tyr Ser Val His Asn305 3 - #10 3 - #15 3 -#20- Gly Asn Phe Gln Thr Trp Met Gly Ala His Ar - #g Ala Gly Val Leu Leu# 335- Ser Gln Asp Cys Ala Gly Thr Pro Gln Gly Al - #a Leu Glu Pro Cys Val# 350- Gln Glu Ala Thr Ala Leu Leu Thr Cys Gly Pr - #o Ala Arg Pro Trp Lys# 365- Ser Val Ala Leu Glu Glu Glu Gln Glu Gly Pr - #o Gly Thr Arg Leu Pro# 380- Gly Asn Leu Ser Ser Glu Asp Val Leu Pro Al - #a Gly Cys Thr Glu Trp385 3 - #90 3 - #95 4 -#00- Arg Val Gln Thr Leu Ala Tyr Leu Pro Gln Gl - #u Asp Trp Ala Pro Thr# 415- Ser Leu Thr Arg Pro Ala Pro Pro Asp Ser Gl - #u Gly Ser Arg Ser Ser# 430- Ser Ser Ser Ser Ser Ser Ser Asn Asn Asn As - #n Tyr Cys Ala Leu Gly# 445- Cys Tyr Gly Gly Trp His Leu Ser Ala Leu Pr - #o Gly Asn Thr Gln Ser# 460- Ser Gly Pro Ile Pro Ala Leu Ala Cys Gly Le - #u Ser Cys Asp His Gln465 4 - #70 4 - #75 4 -#80- Gly Leu Glu Thr Gln Gln Gly Val Ala Trp Va - #l Leu Ala Gly His Cys# 495- Gln Arg Pro Gly Leu His Glu Asp Leu Gln Gl - #y Met Leu Leu Pro Ser# 510- Val Leu Ser Lys Ala Arg Ser Trp Thr Phe# 520- (2) INFORMATION FOR SEQ ID NO: 11:- (i) SEQUENCE CHARACTERISTICS:#acid residuesLENGTH: 307 amino (B) TYPE: amino acid (D) TOPOLOGY: linear#11: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- Met Gly Leu Gly Arg Cys Ile Trp Glu Gly Tr - #p Thr Leu Glu Ser Glu#15- Ala Leu Arg Arg Asp Met Gly Thr Trp Leu Le - #u Ala Cys Ile Cys Ile# 30- Cys Thr Cys Val Cys Leu Gly Val Ser Val Th - #r Gly Glu Gly Gln Gly# 45- Pro Arg Ser Arg Thr Phe Thr Cys Leu Thr As - #n Asn Ile Leu Arg Ile# 60- Asp Cys His Trp Ser Ala Pro Glu Leu Gly Gl - #n Gly Ser Ser Pro Trp# 80- Leu Leu Phe Thr Ser Asn Gln Ala Pro Gly Gl - #y Thr His Lys Cys Ile# 95- Leu Arg Gly Ser Glu Cys Thr Val Val Leu Pr - #o Pro Glu Ala Val Leu# 110- Val Pro Ser Asp Asn Phe Thr Ile Thr Phe Hi - #s His Cys Met Ser Gly# 125- Arg Glu Gln Val Ser Leu Val Asp Pro Glu Ty - #r Leu Pro Arg Arg His# 140- Val Lys Leu Asp Pro Pro Ser Asp Leu Gln Se - #r Asn Ile Ser Ser Gly145 1 - #50 1 - #55 1 -#60- His Cys Ile Leu Thr Trp Ser Ile Ser Pro Al - #a Leu Glu Pro Met Thr# 175- Thr Leu Leu Ser Tyr Glu Leu Ala Phe Lys Ly - #s Gln Glu Glu Ala Trp# 190- Glu Gln Ala Gln His Arg Asp His Ile Val Gl - #y Val Thr Trp Leu Ile# 205- Leu Glu Ala Phe Glu Val Asp Pro Gly Phe Il - #e His Glu Ala Arg Leu# 220- Arg Val Gln Met Ala Thr Leu Glu Asp Asp Va - #l Val Glu Glu Glu Arg225 2 - #30 2 - #35 2 -#40- Tyr Thr Gly Gln Trp Ser Glu Trp Ser Gln Pr - #o Val Cys Phe Gln Ala# 255- Pro Gln Arg Gln Gly Pro Leu Ile Pro Pro Tr - #p Gly Trp Pro Gly Asn# 270- Thr Leu Val Ala Val Ser Ile Phe Leu Leu Le - #u Thr Gly Pro Thr Tyr# 285- Leu Leu Phe Lys Leu Ser Pro Arg Leu Gly Tr - #p Gly Pro Thr Gly Pro# 300- Val Cys Cys305- (2) INFORMATION FOR SEQ ID NO: 12:- (i) SEQUENCE CHARACTERISTICS:#acid residuesLENGTH: 536 amino (B) TYPE: amino acid (D) TOPOLOGY: linear#12: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:- Met Leu Trp Leu Val Val Pro Gly Cys Thr Le - #u His Ser Arg Lys Gly#15- Ala Leu Ser Gln Ser Leu Ala Gly Trp Thr Le - #u Glu Ser Glu Ala Leu# 30- Arg Arg Asp Met Gly Thr Trp Leu Leu Ala Cy - #s Ile Cys Ile Cys Thr# 45- Cys Val Cys Leu Gly Val Ser Val Thr Gly Gl - #u Gly Gln Gly Pro Arg# 60- Ser Arg Thr Phe Thr Cys Leu Thr Asn Asn Il - #e Leu Arg Ile Asp Cys# 80- His Trp Ser Ala Pro Glu Leu Gly Gln Gly Se - #r Ser Pro Trp Leu Leu# 95- Phe Thr Ser Asn Gln Ala Pro Gly Gly Thr Hi - #s Lys Cys Ile Leu Arg# 110- Gly Ser Glu Cys Thr Val Val Leu Pro Pro Gl - #u Ala Val Leu Val Pro# 125- Ser Asp Asn Phe Thr Ile Thr Phe His His Cy - #s Met Ser Gly Arg Glu# 140- Gln Val Ser Leu Val Asp Pro Glu Tyr Leu Pr - #o Arg Arg His Val Lys145 1 - #50 1 - #55 1 -#60- Leu Asp Pro Pro Ser Asp Leu Gln Ser Asn Il - #e Ser Ser Gly His Cys# 175- Ile Leu Thr Trp Ser Ile Ser Pro Ala Leu Gl - #u Pro Met Thr Thr Leu# 190- Leu Ser Tyr Glu Leu Ala Phe Lys Lys Gln Gl - #u Glu Ala Trp Glu Gln# 205- Ala Gln His Arg Asp His Ile Val Glu Val Th - #r Trp Leu Ile Leu Glu# 220- Ala Phe Glu Leu Gln Pro Gly Phe Ile His Gl - #u Ala Arg Leu Arg Val225 2 - #30 2 - #35 2 -#40- Gln Met Ala Thr Leu Gly Asp Asp Val Val Gl - #u Glu Glu Arg Tyr Thr# 255- Gly Gln Trp Ser Glu Trp Ser Gln Pro Val Cy - #s Phe Gln Arg Pro Gln# 270- Arg Gln Gly Pro Leu Ile Pro Pro Trp Gly Tr - #p Pro Gly Asn Thr Leu# 285- Val Ala Val Ser Ile Phe Leu Leu Leu Thr Gl - #y Pro Thr Tyr Leu Leu# 300- Phe Lys Leu Ser Pro Arg Val Lys Arg Ile Ph - #e Tyr Gln Asn Val Pro305 3 - #10 3 - #15 3 -#20- Ser Pro Ala Met Phe Phe Gln Pro Leu Tyr Se - #r Val His Asn Gly Asn# 335- Phe Gln Thr Trp Met Gly Ala His Arg Ala Gl - #y Val Leu Leu Ser Gln# 350- Asp Cys Ala Gly Thr Pro Gln Gly Ala Leu Gl - #y Pro Cys Val Gln Glu# 365- Ala Thr Ala Leu Leu Thr Cys Gly Pro Ala Ar - #g Pro Trp Lys Ser Val# 380- Ala Leu Gly Glu Glu Gln Glu Gly Pro Gly Th - #r Arg Leu Pro Gly Asn385 3 - #90 3 - #95 4 -#00- Leu Ser Ser Glu Asp Val Leu Pro Ala Gly Cy - #s Thr Glu Trp Arg Val# 415- Gln Thr Leu Ala Tyr Leu Pro Gln Glu Asp Tr - #p Ala Pro Thr Ser Leu# 430- Thr Arg Pro Ala Pro Pro Asp Ser Glu Gly Se - #r Arg Ser Ser Ser Ser# 445- Ser Ser Ser Ser Ser Asn Asn Asn Asn Tyr Cy - #s Ala Leu Gly Cys Tyr# 4605- Gly Gly Trp His Leu Ser Ala Leu Pro Gly As - #n Thr Gln Ser Ser Gly465 4 - #70 4 - #75 4 -#80- Pro Ile Pro Ala Leu Ala Cys Gly Leu Ser Cy - #s Asp His Gln Gly Leu# 495- Glu Thr Gln Gln Gly Val Ala Trp Val Leu Al - #a Gly His Cys Gln Arg# 510- Pro Gly Leu His Glu Asp Leu Gln Gly Met Le - #u Leu Pro Ser Val Leu# 525- Ser Lys Ala Arg Ser Trp Thr Phe# 535__________________________________________________________________________

Claims

1. A process for producing an antibody which specifically binds to interleukin 9 receptor comprising immunizing a subject animal with a cell line transfected with a nucleic acid molecule which codes for interleukin-9 receptor in an amount sufficient to generate antibodies which specifically bind to interleukin-9 receptor, and isolating said antibodies from said animal .

2. A method for producing an antibody which specifically binds to interleukin 9 receptor comprising immunizing a non-human animal with a non-glycosylated interleukin 9 receptor in an amount sufficient to generate an antibody specific for interleukin 9 receptor, and purifying said antibody.

3. Monoclonal antibody which specifically binds to an interleukin-9 receptor.

4. The monoclonal antibody of claim 3, which specifically binds to human interleukin-9 receptor.

5. Hybridoma cell line which produces the monoclonal antibody of claim 3.

6. Hybridoma cell line which produces the monoclonal antibody of claim 4.

7. The method according to claim 2 wherein the interleukin 9 receptor is a murine interleukin 9 receptor.

8. The method of claim 1, wherein said cell line is eukaryotic.

9. A method for producing an isolated monoclonal antibody which specifically binds to interleukin 9 receptor comprising immunizing a non-human animal with a protein having an amino acid sequence selected from the group consisting of SEQ ID NO.: 7, 8, 9, 10, 11, and 12, in an amount sufficient to generate a monoclonal antibody specific for interleukin 9 receptor.