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Title:
$i(CHLAMYDIA) ANTIGENS AND CORRESPONDING DNA FRAGMENTS AND USES THEREOF
Document Type and Number:
WIPO Patent Application WO/2000/066739
Kind Code:
A2
Abstract:
The present invention provides a method of nucleic acid, including DNA, immunization of a host, including humans, against disease caused by infection by a strain of $i(Chlamydia), specifically $i(C. pneumoniae), employing a vector containing a nucleotide sequence encoding full-length, 5'-truncated or 3'-truncated 76kDa protein of a strain of $i(Chlamydia pneumoniae) and a promoter to effect expression of the 76kDa protein gene in the host. Modifications are possible within the scope of this invention.

Inventors:
MURDIN ANDREW D (CA)
OOMEN RAYMOND P (CA)
WANG JOE (CA)
DUNN PAMELA (CA)
Application Number:
PCT/CA2000/000511
Publication Date:
November 09, 2000
Filing Date:
May 03, 2000
Export Citation:
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Assignee:
AVENTIS PASTEUR (CA)
MURDIN ANDREW D (CA)
OOMEN RAYMOND P (CA)
WANG JOE (CA)
DUNN PAMELA (CA)
International Classes:
G01N33/53; A61K39/118; A61K39/39; A61K39/395; A61K48/00; A61P9/10; A61P11/00; A61P11/02; A61P11/06; A61P31/04; C07K14/295; C07K16/12; C07K19/00; C12N1/15; C12N1/19; C12N1/21; C12N5/10; C12N15/09; C12N15/31; C12N15/85; C12P21/02; C12Q1/68; G01N33/566; G01N33/569; A61K39/00; (IPC1-7): C12N15/31; C12N15/11; C12N15/62; C12N15/85; C07K14/295; C07K16/12; A61K39/118; A61K31/70
Domestic Patent References:
WO1998058953A21998-12-30
Foreign References:
EP0784059A11997-07-16
Other References:
DATABASE TREMBL [Online] EBI, Hinxton, U.K.; 1 May 1999 (1999-05-01) XP002150537
PEREZ MELGOZA M ET AL: "ISOLATION AND CHARACTERIZATION OF A GENE ENCODING A CHLAMYDIA PNEUMONIAE 76-KILODALTON PROTEIN CONTAINING A SPECIES-SPECIFIC EPITOPE" INFECTION AND IMMUNITY,US,AMERICAN SOCIETY FOR MICROBIOLOGY. WASHINGTON, vol. 62, no. 3, 1 March 1994 (1994-03-01), pages 880-886, XP000198425 ISSN: 0019-9567
See also references of EP 1177301A2
Attorney, Agent or Firm:
Morrow, Joy D. (Ontario K1P 5Y6, CA)
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Claims:
CLAIMS:
1. A nucleic acid molecule comprising a nucleic acid sequence which encodes a polypeptide selected from any of: (a) SEQ ID No: 2; (b) SEQ ID No. 4; (c) SEQ ID No. 6; (d) an immunogenic fragment comprising at least 12 consecutive amino acids from a polypeptide of (a); and (e) a polypeptide of any one of (a) to (d) which has been modified to improve its immunogenicity, wherein said modified polypeptide is at least 75% identical in amino acid sequence to the corresponding polypeptide of any one of (a) to (d).
2. A nucleic acid molecule comprising a nucleic acid sequence selected from any of: (a) SEQ ID No: 1; (b) SEQ ID No: 3; (c) SEQ ID No: 5; (d) a sequence which encodes a polypeptide encoded by any one of SEQ ID Nos: 1,3 and 5; (e) a sequence comprising at least 38 consecutive nucleotides from any one of the nucleic acid sequences of (a) to (d); and (f) a sequence which encodes a polypeptide which is at least 75% identical in amino acid sequence to the polypeptides encoded by any one of SEQ ID Nos: 1,3 and 5.
3. A nucleic acid molecule comprising a nucleic acid sequence which is antisense to the nucleic acid molecule of claim 1.
4. A nucleic acid molecule comprising a nucleic acid sequence which encodes a fusion protein, said fusion protein comprising a polypeptide encoded by a nucleic acid molecule according to claim 1 and an additional polypeptide.
5. The nucleic acid molecule of claim 4 wherein the additional polypeptide is a heterologous signal peptide.
6. The nucleic acid molecule of claim 4 wherein the additional polypeptide has adjuvant activity.
7. A nucleic acid molecule according to any one of claims 1 to 6, operatively linked to one or more expression control sequences.
8. A vaccine comprising at least one first nucleic acid according to any one of claims 1,2, and 4 to 7 and a vaccine vector wherein each first nucleic acid is expressed as a polypeptide, the vaccine optionally comprising a second nucleic acid encoding an additional polypeptide which enhances the immune response to the polypeptide expressed by said first nucleic acid.
9. The vaccine of claim 8 wherein the second nucleic acid encodes an additional Chlamydia polypeptide.
10. A pharmaceutical composition comprising a nucleic acid according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.
11. A pharmaceutical composition comprising a vaccine according to claim 8 or 9 and a pharmaceutically acceptable carrier.
12. A unicellular host transformed with the nucleic acid molecule of claim 7.
13. A nucleic acid probe of 5 to 100 nucleotides which hybridizes under stringent conditions to the nucleic acid molecule of SEQ ID No: 1, or to a homolog or complementary or antisense sequence of said nucleic acid molecule.
14. A primer of 10 to 40 nucleotides which hybridizes under stringent conditions to the nucleic acid molecules of SEQ ID No: 1, or to a homolog or complementary or antisense sequence of said nucleic acid molecule.
15. A polypeptide encoded by a nucleic acid sequence according to any one of claims 1,2 and 4 to 7.
16. A polypeptide comprising an amino acid sequence selected from any of: (a) SEQ ID No: 2; (b) SEQ ID No: 4; (c) SEQ ID No: 6; (d) an immunogenic fragment comprising at least 12 consecutive amino acids from a polypeptide of (a); and (e) a polypeptide of any one of (a) to (d) which has been modified to improve its immunogenicity, wherein said modified polypeptide is at least 75% identical in amino acid sequence to the corresponding polypeptide of any one of (a) to (d).
17. A fusion polypeptide comprising a polypeptide of claim 15 or 16 and an additional polypeptide.
18. The fusion polypeptide of claim 17 wherein the additional polypeptide is a heterologous signal peptide.
19. The fusion protein of claim 17 wherein the additional polypeptide has adjuvant activity.
20. A method for producing a polypeptide of claim 15 or 16, comprising the step of culturing a unicellular host according to claim 12.
21. An antibody against the polypeptide of any one of claims 15 to 19.
22. A vaccine comprising at least one first polypeptide according to any one of claims 15 to 19 and a pharmaceutically acceptable carrier, optionally comprising a second polypeptide which enhances the immune response to the first polypeptide.
23. The vaccine of claim 22 wherein the second polypeptide comprises an additional Chlamydia polypeptide.
24. A pharmaceutical composition comprising a polypeptide according to any one of claims 15 to 19 and a pharmaceutically acceptable carrier.
25. A pharmaceutical composition comprising a vaccine according to claim 22 or 23 and a pharmaceutically acceptable carrier.
26. A pharmaceutical composition comprising an antibody according to claim 21 and a pharmaceutically acceptable carrier.
27. A method for preventing or treating Chlamydia infection using: (a) the nucleic acid of any one of claims 1 to 7; (b) the vaccine of any one of claims 8,9,22 and 23; (c) the pharmaceutical composition of any one of claims 10,11,24 to 26; (d) the polypeptide of any one of claims 15 to 19; or (e) the antibody of claim 21.
28. A method of detecting Chlamydia infection comprising the step of assaying a body fluid of a mammal to be tested, with a component selected from any one of: (a) the nucleic acid of any one of claims 1 to 7; (b) the polypeptide of any one of claims 15 to 19; and (c) the antibody of claim 21.
29. A diagnostic kit comprising instructions for use and a component selected from any one of: (a) the nucleic acid of any one of claims 1 to 7; (b) the polypeptide of any one of claims 15 to 19; and (c) the antibody of claim 21.
30. A method for identifying a polypeptide of claims 15 to 19 which induces an immune response effective to prevent or lessen the severity of Chlamydia infection in a mammal previously immunized with polypeptide, comprising the steps of: (a) immunizing a mouse with the polypeptide; and (b) inoculating the immunized mouse with Chlamydia ; wherein the polypeptide which prevents or lessens the severity of Chlamydia infection in the immunized mouse compare to a nonimmunized control mouse is identified.
31. An expression plasmid selected from the group consisting of pCACPNM555a, pCAI555 and pCAD76kDa.
32. A nucleic acid molecule selected from the group consisting of SEQ ID Nos: 1,3,5 and 7.
33. A polypeptide selected from the group consisting of SEQ ID Nos: 2,4,6 and 8.
34. An isolated 76kDa protein from Chlamydia.
35. An isolated 76kDa protein from Chlamydia pneumoniae.
Description:
INTERNATIONALSEARCHREPORT InternialApplicationNo PCT/CA00/00511 C.(Continuation)DOCUMENTSCONSIDEREDTOBERELEVANT Category°Citationofdocument,withindication,whereappropnate, oftherelevantpassagesRelevanttoclaimNo. AEP0784059A(HITACHICHEMICALCOLTD)1-35 16July1997(1997-07-16) abstract AWO9858953A(MADSENANNASOFIE1-35 ;BIRKELUNDSVEND(DK);KNUDSENKATRINE (DK);MYG)30December1998(1998-12-30) abstract 3 INTERNATIONALSEARCHREPORT Intern lal ApplirationNo ormationonpatentfamilymembers PCT/CA00/00511 PatentdocumentPublication Patent family Publication citedinsearchreportdate member(s)date EP0784059A16-07-1997AU 685680 B 22-01-1998 AU 3532995 A 09-04-1996 WO 9609320 A 28-03-1996 JP 8143594 A 04-06-1996 JP 9009974 A 14-01-1997 JP 9009976 A 14-01-1997 JP 9009999 A 14-01-1997 JP 9015243 A 17-01-1997 JP 9015244 A 17-01-1997 WO9858953A30-12-1998AU 8011998 A 04-01-1999 CN 1261403 T 26-07-2000 EP 1007685 A 14-06-2000