Claims
- 1. A purified polymorphic membrane protein E (PMPE) polypeptide which is encoded by a nucleotide sequence which hybridizes under highly stringent conditions to a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 2 or 73; and which is specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 2 or 73, wherein said highly stringent conditions comprise hybridization at 65° C. in a buffer composed of 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 100 μg/ml denatured salmon sperm DNA, washing at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% BSA, followed by washing at 50° C. in 0.1×SSC for 45 minutes, provided that the purified PMPE polypeptide is not bound specifically by the antibody secreted by hybridoma ATCC No. HB10861.
- 2. The purified PMPE polypeptide of claim 1 which is obtained from a Chlamydia spp.
- 3. The purified PMPE polypeptide of claim 2 wherein the Chlamydia spp. is Chlamydia trachomatis, Chlamydia pneumonia, Chlamydia psittaci or Chlamydia pecorum.
- 4. The purified PMPE polypeptice of claim 2, wherein the Chlamydia spp. is C. trachomatis.
- 5. The purified PMPE polypeptice of claim 1 which has an at least 90% amino acid sequence identity with an amino acid sequence of one of SEQ ID NO.: 2 or 73.
- 6. The purified PMPE polypeptide of claim 1 which has an amino acid sequence of SEQ ID NO.: 2 or 73.
- 7. A purified peptide fragment of the PMPE polypeptide of claim 1, which fragment is at least 8 amino acids in length and which fragment is specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO.: 2 or 73.
- 8. The purified peptide fragment of claim 7 wherein said peptide fragment comprises the amino acid sequence of one of SEQ ID NOs.: 5-22.
- 9. A fusion polypeptide comprising at least two peptides, said at least two peptides each consisting of an amino acid sequence of one of SEQ ID NOs.: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, or 34, with the proviso that the peptides are arranged in a configuration that is different from the configuration of a naturally occurring PMPE or polymorphic membrane protein I (PMPI) polypeptide.
- 10. The fusion polypeptide of claim 9 which is purified.
- 11. An isolated antibody or an antigen-binding fragment thereof that specifically binds the PMPE polypeptide of claim 1.
- 12. An isolated antibody or an antigen-binding fragment thereof that specifically binds a peptide fragment consisting of an amino acid sequence of SEQ ID NO.: 5-22.
- 13. The isolated antibody of claim 11 or 12 which is purified.
- 14. A vaccine comprising the purified PMPE polypeptide of claim 1 and an adjuvant or immunostimulatory compound.
- 15. A vaccine comprising (a) an isolated PMPE polypeptide which is encoded by a nucleotide sequence which hybridizes under highly stringent conditions to a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 2 or 73, and which is specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 2 or 73, wherein said highly stringent conditions comprise hybridization at 65° C. in a buffer composed of 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 100 μg/ml denatured salmon sperm DNA, washing at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% BSA, followed by washing at 50° C. in 0.1×SSC for 45 minutes; (b) an isolated Chlamydia high molecular weight (HMW) protein, polymorphic membrane protein H (PMPH), HtrA protein or major outer membrane protein (MOMP), or an epitope-containing fragment thereof; and (c) and an adjuvant or immunostimulatory compound.
- 16. A vaccine comprising the purified polypeptide fragment of claim 7 and an adjuvant or immunostimulatory compound.
- 17. A vaccine comprising the fusion polypeptide of claim 9 and an adjuvant or immunostimulatory compound.
- 18. The vaccine of claim 17 in which at least one of said peptides is an epitope-containing fragment of a Chlamydia HMW protein, PMPH, HtrA protein or MOMP.
- 19. An isolated nucleic acid molecule comprising a nucleotide sequence encoding a PMPE polypeptide, said nucleotide sequence hybridizing under highly stringent conditions to a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 2 or 73; and said PMPE polypeptide being specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 2 or 73, wherein said highly stringent conditions comprise hybridization at 65° C. in a buffer composed of 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 100 μg/ml denatured salmon sperm DNA, washing at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% BSA, followed by washing at 50° C. in 0.1×SSC for 45 minutes, provided that said PMPE polypeptide is not bound specifically by the antibody secreted from hybridoma ATCC No. HB10861.
- 20. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 1 or 72, or the complement thereof.
- 21. An isolated nucleic acid molecule-comprising a nucleotide sequence encoding an at least 8 amino acid fragment of a PMPE polypeptide, said nucleotide sequence hybridizing under highly stringent conditions to a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 2 or 73; and said fragment being specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 2 or 73, wherein said highly stringent conditions comprise hybridization at 65° C. in a buffer composed of 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 100 μg/ml denatured salmon sperm DNA, washing at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% BSA, followed by washing at 50° C. in 0.1×SSC for 45 minutes.
- 22. The isolated nucleic acid molecule of claim 19, 20, or 21 which is purified.
- 23. A pharmaceutical composition comprising the isolated nucleic acid molecule of claim 19, 20, or 21 and a pharmaceutically acceptable carrier or diluent.
- 24. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the PMPE polypeptide of claim 1.
- 25. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the fusion polypeptide of claim 9.
- 26. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the nucleic acid molecule of claim 19, 20, or 21.
- 27. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the vaccine of claim 14.
- 28. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the vaccine of claim 15.
- 29. Plasmid M15 pREP (pQE-pmpE-Ct-Uni)#37 obtainable from E. coli, as deposited with the ATCC and assigned accession number PTA˜2462.
- 30. A recombinant expression vector adapted for transformation of a host cell comprising the nucleotide sequence of claim 19, 20 or 21.
- 31. The recombinant expression vector of claim 30 further comprising an expression means operatively coupled to the nucleic acid molecule for expression of said nucleic acid by the host cell.
- 32. A transformed host cell containing the recombinant expression vector of claim 31, and progeny of said transformed host cell.
- 33. A host cell containing the nucleotide sequence of claim 19, 20, or 21 operatively linked to a heterologous promoter.
- 34. A purified recombinant PMPE polypeptide produced by a method comprising culturing the host cell of claim 33 under conditions suitable for expression of said PMPE polypeptide and purifying said PMPE polypeptide from said culture of said host cell.
- 35. A method of preventing or treating a disorder or disease associated with infection of an animal with Chlamydia by administering to a subject in need of such prevention or treatment an effective amount of the purified PMPE polypeptide of claim 1.
- 36. A method of preventing or treating a disorder or disease associated with infection of an animal with Chlamydia by administering to a subject in need of such prevention or treatment an effective amount of the vaccine of claim 14.
- 37. A method of preventing or treating a disorder or disease associated with infection of an animal with Chlamydia by administering to a subject in need of such prevention or treatment an effective amount of the vaccine of claim 15.
- 38. A purified PMPI polypeptide which is encoded by a nucleotide sequence which hybridizes under highly stringent conditions to a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 4; and which is specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 4, wherein said highly stringent conditions comprise hybridization at 65° C. in a buffer composed of 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 100 μg/ml denatured salmon sperm DNA, washing at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% BSA, followed by washing at 50° C. in 0.1×SSC for 45 minutes, provided that the purified PMPI polypeptide is not bound specifically by the antibody secreted by hybridoma ATCC No. HB10861.
- 39. The purified PMPI polypeptide of claim 38 which is obtained from a Chlamydia spp.
- 40. The purified PMPI polypeptide of claim 39 wherein the Chlamydia spp. is Chlamydia trachomatis, Chlamydia pneumonia, Chlamydia psittaci or Chlamydia pecorum.
- 41. The purified PMPI polypeptide of claim 39, wherein the Chlamydia spp. is C. trachomatis.
- 42. A purified peptide fragment of the PMPI polypeptide of claim 38, which fragment is at least 8 amino acids in length and which fragment is specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 4.
- 43. The purified peptide fragment of claim 42 wherein said peptide fragment comprises the amino acid sequence of one of SEQ ID NOs.: 23-34.
- 44. An isolated antibody or an antigen-binding fragment thereof that specifically binds the PMPI polypeptide of claim 38.
- 45. A vaccine comprising the purified PNPI polypeptide of claim 38 and an adjuvant or immunostimulatory compound.
- 46. A vaccine comprising (a) an isolated PMPI polypeptide which is encoded by a nucleotide sequence which hybridizes under highly stringent conditions to a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 4, and which is specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 4, wherein said highly stringent conditions comprise hybridization at 65° C. in a buffer composed of 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 100 μg/ml denatured salmon sperm DNA, washing at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% BSA, followed by washing at 50° C. in 0.1×SSC for 45 minutes; (b) an isolated Chlamydia HMW protein, PMPH, HtrA protein or MOMP, or an epitope-containing fragment thereof; and (c) and an adjuvant or immunostimulatory compound.
- 47. An isolated nucleic acid molecule comprising a nucleotide sequence encoding a PMPI polypeptide, said nucleotide sequence hybridizing under highly stringent conditions to a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of SEQ ID NO: 4; and said PMPI polypeptide being specifically bound by an antibody which specifically binds a polypeptide consisting of an amino acid sequence of SEQ ID NO: 4, wherein said highly stringent conditions comprise hybridization at 65° C. in a buffer composed of 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 100 μg/ml denatured salmon sperm DNA, washing at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% BSA, followed by washing at 50° C. in 0.1 ×SSC for 45 minutes, provided that the PMPI is not bound specifically by the antibody secreted by hybridoma ATCC No. HB10861.
- 48. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 3, or the complement thereof.
- 49. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the PMPI polypeptide of claim 38.
- 50. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the nucleic acid molecule of claim 47 or 48.
- 51. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the vaccine of claim 45.
- 52. A method of producing an immune response in an animal comprising administering to the animal an immunogenic amount of the vaccine of claim 46.
- 53. Plasmid TOP10 (pBAD-pmpI-Ct-Uni)#7 obtainable from E. coli, as deposited with the ATCC and assigned accession number PTA˜2461.
- 54. A recombinant expression vector adapted for transformation of a host cell comprising the nucleotide sequence of claim 47 or 48.
- 55. A host cell containing the nucleotide sequence of claim 47 or 48 operatively linked to a heterologous promoter.
- 56. A method of preventing or treating a disorder or disease associated with infection of an animal with Chlamydia by administering to a subject in need of such prevention or treatment an effective amount of the purified PMPI polypeptide of claim 38.
- 57. A method of preventing or treating a disorder or disease associated with infection of an animal with Chlamydia by administering to a subject in need of such prevention or treatment an effective amount of the vaccine of claim 45.
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation-in-part of application Ser. No. 09/677,752, filed Oct. 2, 2000, which is hereby incorporated by reference in its entirety.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/US01/30345 |
9/28/2001 |
WO |
|