NZ520600A - Novel therapeutic compositions for treating infection by Lawsonia spp - Google Patents
Novel therapeutic compositions for treating infection by Lawsonia sppInfo
- Publication number
- NZ520600A NZ520600A NZ520600A NZ52060001A NZ520600A NZ 520600 A NZ520600 A NZ 520600A NZ 520600 A NZ520600 A NZ 520600A NZ 52060001 A NZ52060001 A NZ 52060001A NZ 520600 A NZ520600 A NZ 520600A
- Authority
- NZ
- New Zealand
- Prior art keywords
- plasmid pgte
- nmoo
- plasmid
- pgte
- group
- Prior art date
Links
- 241001469654 Lawsonia <weevil> Species 0.000 title claims abstract 32
- 208000015181 infectious disease Diseases 0.000 title claims abstract 6
- 239000000203 mixture Substances 0.000 title claims 8
- 230000001225 therapeutic effect Effects 0.000 title 1
- 229920001184 polypeptide Polymers 0.000 claims abstract 51
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract 51
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract 51
- 241001148567 Lawsonia intracellularis Species 0.000 claims abstract 33
- 241001239379 Calophysus macropterus Species 0.000 claims abstract 29
- 101100424487 Escherichia coli (strain K12) tamA gene Proteins 0.000 claims abstract 29
- 101150068991 flhB gene Proteins 0.000 claims abstract 29
- 101150065098 glnG gene Proteins 0.000 claims abstract 29
- 101150012415 ntrC gene Proteins 0.000 claims abstract 29
- 101150092975 tlyC gene Proteins 0.000 claims abstract 29
- 101100424491 Escherichia coli (strain K12) tamB gene Proteins 0.000 claims abstract 28
- 230000002163 immunogen Effects 0.000 claims abstract 17
- 229960005486 vaccine Drugs 0.000 claims abstract 13
- 238000000034 method Methods 0.000 claims abstract 11
- 101150026424 fliR gene Proteins 0.000 claims abstract 8
- -1 glnH Proteins 0.000 claims abstract 8
- 101150052329 motA gene Proteins 0.000 claims abstract 8
- 210000001744 T-lymphocyte Anatomy 0.000 claims abstract 7
- 210000003719 b-lymphocyte Anatomy 0.000 claims abstract 7
- 239000013612 plasmid Substances 0.000 claims 188
- 239000002773 nucleotide Substances 0.000 claims 58
- 125000003729 nucleotide group Chemical group 0.000 claims 58
- 125000003275 alpha amino acid group Chemical group 0.000 claims 23
- 108020004414 DNA Proteins 0.000 claims 20
- 210000004027 cell Anatomy 0.000 claims 17
- 108020004707 nucleic acids Proteins 0.000 claims 14
- 102000039446 nucleic acids Human genes 0.000 claims 14
- 150000007523 nucleic acids Chemical class 0.000 claims 14
- 241000271566 Aves Species 0.000 claims 11
- 241001465754 Metazoa Species 0.000 claims 11
- 101150110871 motB gene Proteins 0.000 claims 7
- 241000894006 Bacteria Species 0.000 claims 4
- 150000001413 amino acids Chemical class 0.000 claims 4
- 239000012472 biological sample Substances 0.000 claims 4
- 230000028993 immune response Effects 0.000 claims 4
- 230000001939 inductive effect Effects 0.000 claims 4
- 230000001681 protective effect Effects 0.000 claims 4
- 230000010076 replication Effects 0.000 claims 4
- 239000000523 sample Substances 0.000 claims 4
- 230000000295 complement effect Effects 0.000 claims 3
- 244000005700 microbiome Species 0.000 claims 3
- 210000002966 serum Anatomy 0.000 claims 3
- 241000252983 Caecum Species 0.000 claims 2
- 108091033380 Coding strand Proteins 0.000 claims 2
- 108020004635 Complementary DNA Proteins 0.000 claims 2
- 239000002671 adjuvant Substances 0.000 claims 2
- 239000000427 antigen Substances 0.000 claims 2
- 108091007433 antigens Proteins 0.000 claims 2
- 102000036639 antigens Human genes 0.000 claims 2
- 239000000969 carrier Substances 0.000 claims 2
- 210000004534 cecum Anatomy 0.000 claims 2
- 230000009918 complex formation Effects 0.000 claims 2
- 238000001514 detection method Methods 0.000 claims 2
- 239000003085 diluting agent Substances 0.000 claims 2
- 210000003608 fece Anatomy 0.000 claims 2
- 230000004727 humoral immunity Effects 0.000 claims 2
- 238000009396 hybridization Methods 0.000 claims 2
- 210000003405 ileum Anatomy 0.000 claims 2
- 210000002429 large intestine Anatomy 0.000 claims 2
- 210000001165 lymph node Anatomy 0.000 claims 2
- 238000011321 prophylaxis Methods 0.000 claims 2
- 108090000623 proteins and genes Proteins 0.000 claims 2
- 210000000813 small intestine Anatomy 0.000 claims 2
- 238000011282 treatment Methods 0.000 claims 2
- 108091026890 Coding region Proteins 0.000 claims 1
- 108010006464 Hemolysin Proteins Proteins 0.000 claims 1
- 230000003321 amplification Effects 0.000 claims 1
- 210000004369 blood Anatomy 0.000 claims 1
- 239000008280 blood Substances 0.000 claims 1
- 229940001442 combination vaccine Drugs 0.000 claims 1
- 239000003228 hemolysin Substances 0.000 claims 1
- 230000036039 immunity Effects 0.000 claims 1
- 238000003199 nucleic acid amplification method Methods 0.000 claims 1
- 235000015277 pork Nutrition 0.000 claims 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/205—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Campylobacter (G)
-
- 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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/53—DNA (RNA) vaccination
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
An isolated or recombinant immunogenic polypeptide which comprises, mimics or cross-reacts with a B-cell or T-cell epitope of a Lawsonia spp. (preferably Lawsonia intracellularis) polypeptide selected from the group consisting of flhB, fliR, ntrC, glnH, motA, tlyC, ytfM and ytfN polypeptides; a vaccine comprising said immunogenic polypeptides; antibodies which bind to the polypeptides; and methods of diagnosing infection using the antibodies.
Claims (50)
1. An isolated or recombinant immunogenic polypeptide which comprises, mimics or cross-reacts with a B-cell or T-cell epitope of a Lawsonia spp. polypeptide selected from the group consisting of flhB, fliR, ntrC, glnH, motA, motB, tlyC, ytfM, and ytfN 5 polypeptides.
2. The isolated or recombinant immunogenic polypeptide of claim 1 capable of eliciting the production of antibodies against Lawsonia spp. when administered to an avian or porcine animal.
3. The isolated or recombinant immunogenic polypeptide of claim 1 capable of conferring a protective immune response against Lawsonia spp. when administered to an avian or porcine animal. 15
4. The isolated or recombinant immunogenic polypeptide of claim 2 wherein the Lawsonia spp. is L. intracellularis.
5. The isolated or recombinant immunogenic polypeptide of claim 3 wherein the Lawsonia spp. is L. intracellularis. 20
6. An isolated or recombinant polypeptide selected from the group consisting of: (i) a polypeptide of Lawsonia spp. which comprises an amino acid sequence which has at least about 70% sequence identity overall to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 25 14, 16, and 18; (ii) a polypeptide of Lawsonia spp. which comprises an amino acid sequence which has at least about 70% sequence identity overall to an amino acid sequence encoded by Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 30 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 iponz 11 JUH20M P:\OPER\JMS\SPECIFICATIONS\520600 NZ.DOC- 9/6/04 -84- (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (iii) a polypeptide which comprises at least about 5 contiguous amino acids of an amino acid sequence selected from the group consisting of SEQ ID NOs: 2,4, 6, 8, 10, 12, 14, 16, and 18; (iv) a polypeptide which comprises at least about 5 contiguous amino acids of an amino acid sequence encoded by Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); and (v) a polypeptide which comprises an amino acid sequence encoded by a nucleotide sequence of Lawsonia spp. having at least about 80% sequence identity overall to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17; (vi) a polypeptide which comprises an amino acid sequence encoded by a nucleotide sequence of Lawsonia spp. having at least about 80% sequence identity overall to the nucleotide sequence of Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (vii) a polypeptide encoded by at least about 15 contiguous nucleotides of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5,7,9, 11, 13, 15, and 17; (viii) a polypeptide encoded by at least about 15 contiguous nucleotides of a nucleotide sequence of Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 iponz \\ JUN200*1 WO 02/38594 PCT/AUO1/01462 -85- (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); and (ix) a homologue, analogue or derivative of any one of (i) to (vii) which mimics 5 a B-cell or T-cell epitope of Lawsonia spp.
7. The isolated or recombinant polypeptide of claim 6 capable of eliciting the production of antibodies against Lawsonia spp. in a porcine or avian animal. 10
8. The isolated or recombinant polypeptide of claim 6 capable of conferring a protective immune response against Lawsonia spp. in a porcine or avian animal.
9. The isolated or recombinant polypeptide of claim 8, capable of inducing humoral immunity against Lawsonia spp. in a porcine or avian animal. 15
10. The isolated or recombinant polypeptide of claim 9, capable of inducing humoral immunity against Lawsonia spp. in a porcine animal.
11. The isolated or recombinant polypeptide of claim 7 wherein the Lawsonia spp. 20 is Lawsonia intracellularis.
12. The isolated or recombinant polypeptide of claim 8 wherein the Lawsonia spp. is Lawsonia intracellularis. 25
13. The isolated or recombinant polypeptide of claim 6 comprising an amino acid sequence selected from the group consisting of: (i) an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10,12, 14, 16, and 18; and (ii) an amino acid sequence encoded by Lawsonia intracellularis DNA 30 contained within a deposited plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 WO 02/38594 PCT/AU01/01462 -86- (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN). 5
14. The isolated or recombinant polypeptide of claim 13 capable of eliciting the production of antibodies against Lawsonia intracellularis when administered to an avian or porcine animal.
15. The isolated or recombinant polypeptide of claim 13 capable of inducing a 10 protective immune response against Lawsonia intracellularis in a porcine or avian animal.
16. The isolated or recombinant polypeptide of claim 15 capable of inducing a protective immune response against Lawsonia intracellularis in a porcine animal. 15
17. A vaccine composition for the prophylaxis or treatment of infection of an animal by Lawsonia spp., said vaccine composition comprising an immunogenic component which comprises the isolated or recombinant immunogenic polypeptide according to 20 claim 1 in combination with one or more carriers, diluents or adjuvants suitable for veterinary or pharmaceutical use.
18. The vaccine composition according to claim 17 wherein the Lawsonia spp. is Lawsonia intracellularis. 25
19. The vaccine composition according to claim 17 wherein the immunogenic component comprises an isolated or recombinant polypeptide having an amino acid sequence selected from the group consisting of: (i) an amino acid sequence selected from the group consisting of SEQ 30 ID NOs: 2, 4, 6, 8, 10, 12,14, 16, and 18; and (ii) an amino acid sequence encoded by Lawsonia intracellularis DNA contained within a deposited plasmid selected from the group consisting of P:\OPER\JMS\SPECIFICATIONS\52O6OO NZ.DOC - 9/6/04 -87- AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN);
20. The vaccine composition of claim 17, wherein the immunogenic component is a recombinant polypeptide expressed in a cell that has been transfected with a vector comprising a nucleotide sequence selected from the group consisting of: (i) a protein-encoding nucleotide sequence having at least about 80% sequence identity overall to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17; (ii) a protein-encoding nucleotide sequence having at least about 80% sequence identity overall to the protein-encoding sequence of Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (iii) a protein-encoding nucleotide sequence which comprises at least about 15 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9,11,13, 15, and 17; (iv)a protein-encoding nucleotide sequence which comprises at least about 15 contiguous nucleotides of the protein-encoding sequence of Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (v) a protein-encoding nucleotide sequence which hybridizes under at least low iponZ P:\OPER\JMS\SPECIFICATIONS\520600 NZ.DOC- 9/6/04 -88- stringency conditions to the complement of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1,3,5,7,9,11,13,15, and 17; (vi)a protein-encoding nucleotide sequence which hybridizes under at least low stringency conditions to the non-coding strand of Lawsonia intracellularis DNA 5 contained within a p plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 10 (plasmid pGTE#8 ytfN); and (vii) a homologue, analogue or derivative of any one of (i) to (vi) which encodes a polypeptide which mimics a B-cell or T-cell epitope of Lawsonia spp.
21. A combination vaccine composition for the prophylaxis or treatment of infection 15 of an animal by Lawsonia spp., said vaccine composition comprising: (i) a first immunogenic component which comprises the isolated or recombinant polypeptide having according to claim 1; (ii) a second immunogenic component different from said first immunogenic component and comprising a polypeptide selected from the group consisting 20 of the Lawsonia intracellularis FlgE, hemolysin, OmpH, SodC, flhB, fliR, ntrC, glnH, motA, motB, tlyC, ytfM, and ytfN polypeptides; and (iii) one or more carriers, diluents or adjuvants suitable for veterinary or pharmaceutical use. 25
22. A vaccine vector that comprises, in an expressible form, an isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: (i) a protein-encoding nucleotide sequence having at least about 80% sequence identity overall to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17; 30 (ii) a protein-encoding nucleotide sequence having at least about 80% sequence identity overall to the protein-encoding sequence of Lawsonia intracellularis DNA contained within a plasmid selected from the group iponz \\ JUW 20M WO 02/38594 PCT/AU01/01462 - 89 - consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); 5 and NM01/23286 (plasmid pGTE#8 ytfN); (iii) a protein-encoding nucleotide sequence which comprises at least about 15 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11,13,15, and 17; (iv)a protein-encoding nucleotide sequence which comprises at least about 15 10 contiguous nucleotides of the protein-encoding sequence of Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); 15 NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (v) a protein-encoding nucleotide sequence which hybridizes under at least low stringency conditions to the complement of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9,11, 13, 15, and 17; 20 (vi)a protein-encoding nucleotide sequence which hybridizes under at least low stringency conditions to the non-coding strand of Lawsonia intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid 25 pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); and (vii) a homologue, analogue or derivative of any one of (i) to (vi) which enqodes a polypeptide which mimics a B-cell or T-cell epitope of Lawsonia spp. 30 wherein said vaccine vector expresses the polypeptide encoded by said nucleotide sequence at a level sufficient to confer immunity against Lawsonia spp. when administered to a porcine or avian animal. P:\OPER\JMS\SPECIFICATIONS\520600 NZ.DOC - 9/6/04 -90-
23. The vaccine vector of claim 22 wherein the immunogenic polypeptide is expressed by a process comprising: (i) placing an isolated nucleic acid molecule in an expressible format, said nucleic acid molecule comprising the coding region of a gene selected from the group consisting of flhB, fliR, ntrC, glnH, motA, motB, tlyC, ytfM, and ytfN genes, or a protein-encoding homologue, analogue or derivative thereof; (ii) introducing the isolated nucleic acid molecule of (i) in an expressible format into a suitable vaccine vector; and (iii) incubating or growing the vaccine vector for a time and under conditions sufficient for expression of the immunogenic component encoded by said nucleic acid molecule to occur.
24. The vaccine vector of claim 22 wherein the Lawsonia spp. is L. intracellularis. 15
25. An isolated polyclonal antibody or a monoclonal antibody molecule that binds specifically to a Lawsonia spp. polypeptide selected from the group consisting of flhB, fliR, ntrC, glnH, motA, motB, tlyC, ytfM, and ytfN polypeptides, or a homologue, analogue or derivative of any one or more of said polypeptides. 20
26. The isolated polyclonal antibody or a monoclonal antibody molecule of claim 25 wherein the polypeptide or derivative thereof comprises an amino acid sequence selected from the group consisting of; (i) an amino acid sequence of Lawsonia sp. which has at least about 70% 25 sequence identity overall to a sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, and 18; (ii) an amino acid sequence of Lawsonia sp. which has at least about 70% sequence identity overall to a sequence encoded by L. intracellularis DNA contained within a plasmid selected from the group consisting of AGAL 30 Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid iponz \\ jUH2001! 10 P:\OPER\JMS\SPECIFICATIONS\52O6OO NZ.DOC- 9/6/04 -91 - pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (iii) an amino acid sequence which comprises at least about 5 contiguous amino acids of a sequence selected from the group consisting of SEQ ID NOs: 2, 4, 5 6, 8, 10, 12, 14, 16, and 18; (iv)an amino acid sequence which comprises at least about 5 contiguous amino acids of a sequence encoded by L. intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB) 10 NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B) NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC) NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (v) an amino acid sequence which is encoded by a nucleotide sequence 15 having at least about 80% sequence identity overall to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17; (vi)an amino acid sequence which is encoded by a nucleotide sequence having at least about 80% sequence identity overall to the nucleotide sequence of L 20 intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH) NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR) NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC) NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM) 25 and NM01/23286 (plasmid pGTE#8 ytfN); (vii) an amino acid sequence which is encoded by at least about 15 contiguous nucleotides of a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17; (viii) an amino acid sequence which is encoded by at least about 15 30 contiguous nucleotides of a nucleotide sequence of L. intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid iponz 11 JUW20W P:\OPER\JMS\SPECIFICATIONS\52O6OO NZ.DOC- 9/6/04 -92- pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); and 5 (ix) a homologue, analogue or derivative of any one of (i) to (viii) which mimics a B-cell or T-cell epitope of Lawsonia spp.
27. A method of diagnosing infection of a porcine or avian animal by Lawsonia intracellularis or a microorganism that is immunologically cross-reactive thereto, said 10 method comprising the steps of contacting a biological sample derived from said animal with the antibody molecule of claim 25 for a time and under conditions sufficient for an antigen:antibody complex to form, and then detecting said complex formation. 15
28. The method of claim 27 wherein the biological sample comprises whole serum, lymph nodes, ileum, caecum, small intestine, large intestine, faeces or a rectal swab derived from a porcine animal.
29. A method of identifying whether or not a porcine or avian animal has suffered 20 from a past infection, or is currently infected, with Lawsonia intracellularis or a microorganism that is immunologically cross-reactive thereto, said method comprising contacting blood or serum derived from said animal with the immunogenic polypeptide of claim 1 for a time and under conditions sufficient for an antigen:antibody complex to form and then detecting said complex formation. 25
30. An isolated nucleic acid molecule which consists of a nucleotide sequence encoding a Lawsonia spp. polypeptide selected from the group consisting of flhB, fliR, ntrC, glnH, motA, motB, tlyC, ytfM, and ytfN. 30
31. The isolated nucleic acid molecule according to claim 30 comprising a sequence of nucleotides selected from the group consisting of: (i) a nucleotide sequence having at least about 80% sequence identity overall iponz w m 20M P:\OPER\JMS\SPECIFICATIONS\520600 NZ.DOC • 9/6/04 -93- to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17; (ii) a nucleotide sequence having at least about 80% sequence identity overall to L. intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NMOO/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and; (iii) a nucleotide sequence which comprises at least about 15 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1,3, 5, 7, 9, 11, 13, 15, and 17; (iv)a nucleotide sequence which comprises at least about 15 contiguous nucleotides of L. intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NMOO/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (v) a nucleotide sequence which hybridizes under at least low stringency conditions to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17 or a complementary nucleotide sequence thereto; (vi)a nucleotide sequence which hybridizes under at least low stringency conditions to L. intracellularis DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NMOO/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); and (vii) a homologue, analogue or derivative of any one of (i) to (vi) which encodes a polypeptide which mimics a B-cell or T-cell epitope of Lawsonia spp. \pouz 11 JUW20M WO 02/38594 PCT/AU01/01462 -94-
32. The isolated nucleic acid molecule of claim 31 comprising a nucleotide sequence selected from the group consisting of: (i) a nucleotide sequence selected from the group consisting of SEQ ID NOs: 5 1,3, 5,7, 9,11,13, 15, and 17; (ii) a nucleotide sequence of the L. intracellularis DNA contained within a deposited plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NMOO/16479 (plasmid pGTE#4 motA/B); 10 NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN); (iii) a nucleotide sequence that encodes the same polypeptide as a nucleotide sequence of (i) or (ii), wherein said polypeptide is selected from the group 15 consisting of flhB, fliR, ntrC, glnH, motA, motB, tlyC, ytfM, and ytfN polypeptides; and (iv)a nucleotide sequence that is complementary to (i) or (ii) or (iii).
33. The isolated nucleic acid molecule of claim 32 consisting of the protein-20 encoding region of (i) or (ii).
34. A method of detecting Lawsonia intracellularis or related microorganism in a biological sample derived from a porcine or avian animal subject, said method comprising the steps of hybridising one or more probes or primers to said sample and 25 then detecting said hybridisation using a detection means, wherein said probes or primers are derived from a Lawsonia spp. gene selected from the group consisting of flhB, fliR, ntrC, glnH, motA, motB, tlyC, ytfM, and ytfN genes.
35. The method of claim 34 wherein the biological sample comprises whole serum, 30 lymph nodes, ileum, caecum, small intestine, large intestine, faeces or a rectal swab derived from a porcine animal. P:\OPER\JMS\SPECIFICAT10NS\520600 NZ.DOC • 9/6/04 -95-
36. The method of claim 34 wherein the detection means comprises any nucleic acid based hybridisation or amplification reaction.
37. A probe or primer comprising a nucleotide sequence selected from the group 5 consisting of: (i) any one of SEQ ID NOs: 19 to 68; and (ii) a complementary nucleotide sequence to (i).
38. A plasmid selected from the group consisting of AGAL Accession Nos: NMOO/16476 (plasmid pGTE#1 glnH); NMOO/16477 (plasmid pGTE#2 flhB); NMOO/16478 (plasmid pGTE#3 fliR); NMOO/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NMOO/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN).
39. A recombinant vector capable of replication in a host cell, wherein said vector comprises the isolated nucleic acid of claim 30.
40. A recombinant vector capable of replication in a host cell, wherein said vector comprises the isolated nucleic acid of claim 31.
41. A recombinant vector capable of replication in a host cell, wherein said vector comprises the isolated nucleic acid of claim 32. 25
42. A recombinant vector capable of replication in a host cell, wherein said vector comprises the isolated nucleic acid of claim 33.
43. An isolated host cell comprising the recombinant vector of claim 39. 30
44. The host cell of claim 43 wherein said host cell is a bacterium.
45. An isolated host cell comprising the recombinant vector of claim 40. iponz JUN 2001! P:\OPER\JMS\SPECIFICATIONS\520600 NZ.DOC- 9/6/04 -96-
46. The host cell of claim 45 wherein said host cell is a bacterium.
47. An isolated host cell comprising the recombinant vector of claim 41.
48. The host cell of claim 47 wherein said host cell is a bacterium.
49. An isolated host cell comprising the recombinant vector of claim 42.
50. The host cell of claim 49 wherein said host cell is a bacterium. Agriculture Victoria Services Pty. Ltd., Australian Pork Limited AND Pfizer Products, Inc. By their Attorneys Davies Collison Cave \P0W2 \\ JUU200H
Applications Claiming Priority (3)
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AUPR1381A AUPR138100A0 (en) | 2000-11-10 | 2000-11-10 | Novel therapeutic compositions for treating infection by lawsonia spp |
US24959600P | 2000-11-17 | 2000-11-17 | |
PCT/AU2001/001462 WO2002038594A1 (en) | 2000-11-10 | 2001-11-09 | Novel therapeutic compositions for treating infection by lawsonia spp |
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NZ520600A NZ520600A (en) | 2000-11-10 | 2001-11-09 | Novel therapeutic compositions for treating infection by Lawsonia spp |
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EP (1) | EP1332154A4 (en) |
JP (1) | JP2004512851A (en) |
BR (1) | BR0114835A (en) |
CA (1) | CA2399276A1 (en) |
MX (1) | MXPA03003077A (en) |
NZ (1) | NZ520600A (en) |
WO (1) | WO2002038594A1 (en) |
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AU2003295341A1 (en) * | 2002-10-04 | 2004-05-04 | Regents Of The University Of Minnesota | Nucleic acid and polypeptide sequences from lawsonia intracellularis and methods of using |
US8834891B2 (en) | 2005-03-14 | 2014-09-16 | Boehringer Ingelheim Vetmedica, Inc. | Immunogenic compositions comprising Lawsonia intracellularis |
US8398994B2 (en) | 2005-07-15 | 2013-03-19 | Boehringer Ingelheim Vetmedica, Inc. | Lawsonia vaccine and methods of use thereof |
US20100129397A1 (en) | 2006-12-11 | 2010-05-27 | Boehringer Ingelheim Vetmedica, Inc. | Effective method of treatment of porcine circovirus and lawsonia intracellularis infections |
FR2980213B1 (en) * | 2011-09-16 | 2013-09-06 | Biomerieux Sa | PROCESS FOR CHARACTERIZING BACTERIA BY DETECTION OF NON-STRUCTURAL PROTEINS OF BACTERIOPHAGES |
US9359643B2 (en) | 2013-03-08 | 2016-06-07 | Progenika Biopharma S.A. | Discrimination of blood type variants |
CL2017002196A1 (en) | 2017-08-30 | 2017-11-17 | Univ De Concepción | Recombinant vaccine against proliferative enteropathy in animals. |
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US5885823A (en) * | 1995-06-05 | 1999-03-23 | Nobl Laboratories, Inc. | Lawsonia intracellularis cultivation, anti-Lawsonia intracellularis vaccines and diagnostic agents |
JP2000502054A (en) * | 1995-11-30 | 2000-02-22 | アグリカルチャー ビクトリア サービシーズ ピーティーワイ.エルティーディー. | Therapeutic and diagnostic compositions |
WO1999058682A2 (en) * | 1998-05-12 | 1999-11-18 | Smithkline Beecham Biologicals S.A. | Compounds from moraxella catarrhalis |
TWI224107B (en) * | 1998-10-22 | 2004-11-21 | Pfizer Prod Inc | Novel proteins from actinobacillus pleuropneumoniae |
GB9902880D0 (en) * | 1999-02-09 | 1999-03-31 | Smithkline Beecham Biolog | Novel compounds |
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2001
- 2001-11-09 CA CA002399276A patent/CA2399276A1/en not_active Abandoned
- 2001-11-09 MX MXPA03003077A patent/MXPA03003077A/en unknown
- 2001-11-09 WO PCT/AU2001/001462 patent/WO2002038594A1/en not_active Application Discontinuation
- 2001-11-09 NZ NZ520600A patent/NZ520600A/en unknown
- 2001-11-09 JP JP2002541925A patent/JP2004512851A/en active Pending
- 2001-11-09 BR BR0114835-4A patent/BR0114835A/en not_active IP Right Cessation
- 2001-11-09 EP EP01983297A patent/EP1332154A4/en not_active Withdrawn
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MXPA03003077A (en) | 2003-09-25 |
BR0114835A (en) | 2003-07-01 |
JP2004512851A (en) | 2004-04-30 |
WO2002038594A1 (en) | 2002-05-16 |
CA2399276A1 (en) | 2002-05-16 |
EP1332154A4 (en) | 2005-05-11 |
EP1332154A1 (en) | 2003-08-06 |
WO2002038594A9 (en) | 2002-11-07 |
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