WO2001021810B1 - Chlamydia antigens and corresponding dna fragments and uses thereof - Google Patents
Chlamydia antigens and corresponding dna fragments and uses thereofInfo
- Publication number
- WO2001021810B1 WO2001021810B1 PCT/CA2000/001085 CA0001085W WO0121810B1 WO 2001021810 B1 WO2001021810 B1 WO 2001021810B1 CA 0001085 W CA0001085 W CA 0001085W WO 0121810 B1 WO0121810 B1 WO 0121810B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- polypeptide
- nucleic acid
- seq
- vaccine
- acid sequence
- Prior art date
Links
- 241000606161 Chlamydia Species 0.000 title claims abstract 6
- 239000012634 fragment Substances 0.000 title claims 19
- 239000000427 antigen Substances 0.000 title 1
- 102000036639 antigens Human genes 0.000 title 1
- 108091007433 antigens Proteins 0.000 title 1
- 150000007523 nucleic acids Chemical class 0.000 claims abstract 49
- 108020004707 nucleic acids Proteins 0.000 claims abstract 33
- 102000039446 nucleic acids Human genes 0.000 claims abstract 33
- 239000002773 nucleotide Substances 0.000 claims abstract 8
- 125000003729 nucleotide group Chemical group 0.000 claims abstract 8
- 230000000694 effects Effects 0.000 claims abstract 5
- 238000000034 method Methods 0.000 claims abstract 4
- 241001647372 Chlamydia pneumoniae Species 0.000 claims abstract 3
- 230000003053 immunization Effects 0.000 claims abstract 2
- 208000015181 infectious disease Diseases 0.000 claims abstract 2
- 229920001184 polypeptide Polymers 0.000 claims 91
- 108090000765 processed proteins & peptides Proteins 0.000 claims 91
- 102000004196 processed proteins & peptides Human genes 0.000 claims 91
- 229960005486 vaccine Drugs 0.000 claims 24
- 108020001507 fusion proteins Proteins 0.000 claims 17
- 102000037865 fusion proteins Human genes 0.000 claims 17
- 108091028043 Nucleic acid sequence Proteins 0.000 claims 14
- 125000003275 alpha amino acid group Chemical group 0.000 claims 12
- 230000002163 immunogen Effects 0.000 claims 10
- 230000028993 immune response Effects 0.000 claims 7
- 150000001413 amino acids Chemical class 0.000 claims 6
- 230000005847 immunogenicity Effects 0.000 claims 6
- 239000008194 pharmaceutical composition Substances 0.000 claims 6
- 239000003937 drug carrier Substances 0.000 claims 5
- 108010076504 Protein Sorting Signals Proteins 0.000 claims 4
- 239000002671 adjuvant Substances 0.000 claims 4
- 208000007190 Chlamydia Infections Diseases 0.000 claims 3
- 230000000692 anti-sense effect Effects 0.000 claims 3
- 241000124008 Mammalia Species 0.000 claims 2
- 230000000295 complement effect Effects 0.000 claims 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims 1
- 238000009007 Diagnostic Kit Methods 0.000 claims 1
- 108020004711 Nucleic Acid Probes Proteins 0.000 claims 1
- 210000001124 body fluid Anatomy 0.000 claims 1
- 239000010839 body fluid Substances 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- 239000013613 expression plasmid Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 239000002853 nucleic acid probe Substances 0.000 claims 1
- 101150103033 lpxB gene Proteins 0.000 abstract 2
- -1 DNA Chemical class 0.000 abstract 1
- 201000010099 disease Diseases 0.000 abstract 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract 1
- 238000002649 immunization Methods 0.000 abstract 1
- 238000012986 modification Methods 0.000 abstract 1
- 230000004048 modification Effects 0.000 abstract 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1051—Hexosyltransferases (2.4.1)
- C12N9/107—1,4-Alpha-glucan branching enzyme (2.4.1.18)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
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 Chlamydia, specifically C. pneumoniae, employing a vector containing a nucleotide sequence encoding lpxB of a strain of Chlamydia pneumoniae and a promoter to effect expression of the lpxB gene in the host. Modifications are possible within the scope of this invention.
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) an immunogenic fragment comprising at least 12 consecutive amino acids from a polypeptide of (a) ; and
(c) a polypeptide of (a) or (b) which has been modified without loss of immunogenicity, wherein said modified polypeptide is at least 75% identical in amino acid sequence to the corresponding polypeptide of (a) or (b) .
2. A nucleic acid molecule comprising a nucleic acid sequence selected from any of :
(a) SEQ ID No: 1;
(b) a sequence which encodes a polypeptide encoded by SEQ ID No: 1;
(c) a sequence comprising at least 38 consecutive nucleotides from any one of the nucleic acid sequences of (a) and (b) ; and
(d) a sequence which encodes a polypeptide which is at least 75% identical in amino acid sequence to the polypeptide encoded by SEQ ID No: 1.
3. A nucleic acid molecule comprising a nucleic acid sequence which is anti-sense to the nucleic acid molecule of claim 1 or 2.
4. A nucleic acid molecule comprising a nucleic acid sequence which encodes a fusion protein, said fusion protein
59
comprising a polypeptide encoded by a nucleic acid molecule according to claim 1 and a second polypeptide.
5. The nucleic acid molecule of claim 4 wherein the second polypeptide is a heterologous signal peptide.
6. The nucleic acid molecule of claim 4 wherein the second 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 a vaccine vector and at least one first nucleic acid selected from any of:
(i) SEQ ID No: 1;
(ii) a nucleic acid sequence which encodes a polypeptide encoded by SEQ ID No: 1;
(iii) a nucleic acid sequence comprising at least 38 consecutive nucleotides from any one of the nucleic acid sequences of (i) and (ii) ;
(iv) a nucleic acid sequence which encodes a polypeptide which is at least 75% identical in amino acid sequence to the polypeptide encoded by SEQ ID No: 1;
(v) a nucleic acid sequence which encodes a polypeptide whose sequence is set forth in SEQ ID No: 2;
(vi) a nucleic acid sequence which encodes an immunogenic fragment comprising at least 12 consecutive amino acids from SEQ ID No: 2; and
(vii) a nucleic acid sequence which encodes a polypeptide as defined in (i) to (v) or an immunogenic fragment
60
as defined in (vi) which has been modified without loss of immunogenicity, wherein said modified polypeptide or fragment is at least 75% identical in amino acid sequence to the corresponding polypeptide of (i) to (v) or the corresponding fragment of (vi) ;
wherein each first nucleic acid is capable of being expressed and wherein the vaccine optionally comprises a second nucleic acid encoding and capable of expressing an additional polypeptide which enhances the immune response to the polypeptide expressed by the first nucleic acid.
9. A vaccine comprising a vaccine vector and at least one first nucleic acid encoding a fusion protein, wherein the fusion protein comprises:
(a) a first polypeptide selected from any of:
(i) a polypeptide encoded by SEQ ID No: 1;
(ii) a polypeptide encoded by a nucleic acid sequence comprising at least 38 consecutive nucleotides from SEQ ID NO: 1;
(iϋ) a polypeptide which is at least 75% identical in amino acid sequence to the polypeptide encoded by SEQ ID No: l;
(iv) a polypeptide whose sequence is set forth in SEQ ID No: 2 ;
(v) an immunogenic fragment comprising at least 12 consecutive amino acids from SEQ ID No: 2; and
61
(vi) a polypeptide as defined in (i) to (iv) or an immunogenic fragment as defined in (v) which has been modified without loss of immunogenicity, wherein said modified polypeptide or fragment is at least 75% identical in amino acid sequence to the corresponding polypeptide of (i) to (iv) or the corresponding fragment of (v) ; and
(b) a second polypeptide;
wherein each first nucleic acid is capable of being expressed and wherein the vaccine optionally comprises a second nucleic acid encoding and capable of expressing an additional polypeptide which enhances the immune response to the first polypeptide.
10. The vaccine of claim 9 wherein the second polypeptide is a heterologous signal peptide.
11. The vaccine of claim 9 wherein the second polypeptide has adjuvant activity.
12. The vaccine of any one of claims 8 to 11 wherein wherein each first nucleic acid is operatively linked to one or more expression control sequences.
13. 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.
14. The vaccine of any one of claims 8 to 13 wherein the second nucleic acid encodes an additional Chlamydia polypeptide.
15. A pharmaceutical composition comprising a nucleic acid according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.
16. A pharmaceutical composition comprising a vaccine according to any one of claims 8 to 14 and a pharmaceutically acceptable carrier.
17. A unicellular host transformed with the nucleic acid molecule of claim 7.
18. An isolated 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 complementary or anti-sense sequence of said nucleic acid molecule.
19. An isolated primer of 10 to 40 nucleotides which hybridizes under stringent conditions to the nucleic acid molecules of SEQ ID No: 1, or to a complementary or anti-sense sequence of said nucleic acid molecule.
20. A polypeptide encoded by a nucleic acid sequence according to any one of claims 1, 2 and 4 to 7.
21. A polypeptide comprising an amino acid sequence selected from any of:
(a) SEQ ID No: 2;
(b) an immunogenic fragment comprising at least 12 consecutive amino acids from a polypeptide of (a) ,- and
(c) a polypeptide of (a) or (b) which has been modified without loss of immunogenicity, wherein said modified polypeptide is at least 75% identical in amino acid sequence to the corresponding polypeptide of (a) or (b) .
6 3
22. A fusion protein comprising a polypeptide of claim 20 or 21 and a second polypeptide.
23. The fusion protein of claim 22 wherein the second polypeptide is a heterologous signal peptide.
24. The fusion protein of claim 22 wherein the second polypeptide has adjuvant activity.
25. A method for producing a polypeptide of claim 20 or 21, or a fusion protein of any one of claims 22 to 24, comprising the step of culturing a unicellular host of claim 17.
26. An antibody against the polypeptide of claim 20 or 21, or against a fusion protein of any one of claims 22 to 24.
27. A vaccine comprising at least one first polypeptide selected from any o :
(i) a polypeptide encoded by SEQ ID No: 1;
(ii) a polypeptide encoded by a nucleic acid sequence comprising at least 38 consecutive nucleotides from SEQ ID No: 1;
(iii) a polypeptide which is at least 75% identical in amino acid sequence to the polypeptide encoded by SEQ ID No: 1;
(iv) a polypeptide whose sequence is set forth in SEQ ID No: 2 ;
(v) an immunogenic fragment comprising at least 12 consecutive amino acids from SEQ ID No: 2; and
(vi) a polypeptide as defined in (i) to (iv) or an immunogenic fragment as defined in (v) which has been modified
6 4
without loss of immunogenicity, wherein said modified polypeptide or fragment is at least 75% identical in amino acid sequence to the corresponding polypeptide of (i) to (iv) or the corresponding fragment of (v) ;
wherein the vaccine optionally comprises an additional polypeptide which enhances the immune response to the first polypeptide.
28. A vaccine comprising at least one fusion protein, wherein the fusion protein comprises:
(a) a first polypeptide selected from any of:
(i) a polypeptide encoded by SEQ ID No: 1;
(ii) a polypeptide encoded by a nucleic acid sequence comprising at least 38 consecutive nucleotides from SEQ ID No: 1;
(iii) a polypeptide which is at least 75% identical in amino acid sequence to the polypeptide encoded by SEQ ID No: 1;
(iv) a polypeptide whose sequence is set forth in SEQ ID No: 2 ;
(v) an immunogenic fragment comprising at least 12 consecutive amino acids from SEQ ID No:2; and
(vi) a polypeptide as defined in (i) to (iv) or an immunogenic fragment as defined in (v) which has been modified without loss of immunogenicity, wherein said modified polypeptide or fragment is at least 75% identical in amino acid sequence to the corresponding polypeptide of (i) to (iv) or the corresponding fragment of (v) ; and
65
(b) a second polypeptide;
wherein the vaccine optionally comprises an additional polypeptide which enhances the immune response to the first polypeptide.
29. The vaccine of claim 28 wherein the second polypeptide is a heterologous signal peptide.
30. The vaccine of claim 28 wherein the second polypeptide has adjuvant activity.
31. A vaccine comprising at least one first polypeptide according to any one of claims 20 to 24, optionally comprising an additional polypeptide which enhances the immune response to the first polypeptide.
32. The vaccine of any one of claims 27 to 31 wherein the additional polypeptide comprises a Chlamydia polypeptide.
33. A pharmaceutical composition comprising a polypeptide according to any one of claims 20 to 24 and a pharmaceutically acceptable carrier.
34. A pharmaceutical composition comprising a vaccine according to any one of claims 27 to 32 and a pharmaceutically acceptable carrier.
35. A pharmaceutical composition comprising an antibody according to claim 26 and a pharmaceutically acceptable carrier.
36. A method for preventing or treating Chla nydia infection using:
(a) the nucleic acid of any one of claims 1 to 7;
66
(b) the vaccine of any one of claims 8 to 14 and 27 to 32;
(c) the pharmaceutical composition of any one of claims 15, 16 and 33 to 35;
(d) the polypeptide of claim 20 or 21, or a fusion protein of any one of claims 22 to 24; or
(e) the antibody of claim 26.
37. 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 claim 20 or 21, or a fusion protein of any one of claims 22 to 24; and
(c) the antibody of claim 26.
38. 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 claim 20 or 21, or a fusion protein of any one of claims 22 to 24; and
(c) the antibody of claim 26.
39. A method for identifying a polypeptide of claim 20 or 21, or a fusion protein of any one of claims 22 to 24 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:
6 7
(a) immunizing a mouse with the polypeptide or fusion protein; and
(b) inoculating the immunized mouse with Chlamydia;
wherein the polypeptide or fusion protein which prevents or lessens the severity of Chlamydia infection in the immunized mouse compared to a non-immunized control mouse is identified.
40. Expression plasmid pCABkl043 as shown in Figure 3.
41. A nucleic acid molecule of SEQ ID NO. 3 or 4.
42. An isolated IpxB polypeptide from Chlamydia .
43. An isolated IpxB polypeptide from Chlamydia pneumoniae .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU73982/00A AU7398200A (en) | 1999-09-17 | 2000-09-15 | Chlamydia antigens and corresponding dna fragments and uses thereof |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US15446199P | 1999-09-17 | 1999-09-17 | |
| US60/154,461 | 1999-09-17 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO2001021810A1 WO2001021810A1 (en) | 2001-03-29 |
| WO2001021810B1 true WO2001021810B1 (en) | 2002-06-06 |
Family
ID=22551451
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CA2000/001085 WO2001021810A1 (en) | 1999-09-17 | 2000-09-15 | Chlamydia antigens and corresponding dna fragments and uses thereof |
Country Status (2)
| Country | Link |
|---|---|
| AU (1) | AU7398200A (en) |
| WO (1) | WO2001021810A1 (en) |
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|---|---|---|---|---|
| JP2002536958A (en) * | 1997-11-21 | 2002-11-05 | ジェンセット | Chlamydiapneumoniae genomic sequences and polypeptides, fragments thereof, and their use especially for the diagnosis, prevention and treatment of infectious diseases |
-
2000
- 2000-09-15 AU AU73982/00A patent/AU7398200A/en not_active Abandoned
- 2000-09-15 WO PCT/CA2000/001085 patent/WO2001021810A1/en active Application Filing
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Also Published As
| Publication number | Publication date |
|---|---|
| WO2001021810A1 (en) | 2001-03-29 |
| AU7398200A (en) | 2001-04-24 |
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