Novel therapeutic compositions for treating infection by Lawsonia spp.

Abstract

The present invention relates generally to therapeutic compositions for the treatment and/or prophylaxis of intestinal disease conditions in animals and birds caused or exacerbated by Lawsonia intracellularis or similar or otherwise related microorganism. In particular, the present invention provides a novel gene derived from Lawsonia intracellularis , which encodes an immunogenic polypeptide that is particularly useful as an antigen in a vaccine preparation for conferring humoral immunity against Lawsonia intracellularis and related pathogens in animal hosts, wherein said polypeptide is 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. The present invention is also directed to methods for the treatment and/or prophylaxis of such intestinal disease conditions and to diagnostic agents and procedures for detecting Lawsonia intracellularis or similar or otherwise related microorganisms.

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 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.  
     
     
         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.    
     
     
         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 60% sequence identity overall to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, and 18;    (ii) a polypeptide of  Lawsonia  spp. which comprises an amino acid sequence which has at least about 60% 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: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (plasmid pGTE#6 ntrC); NM00/16482 (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: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (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 60% 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 60% sequence identity overall to the nucleotide sequence of  Lawsonia intracellularis  DNA contained within a plasmid selected from the group consisting of AGAL Accession Nos: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (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: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motB); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (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 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.  
     
     
         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.  
     
     
         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. is  Lawsonia intracellularis.    
     
     
         12 . The isolated or recombinant polypeptide of  claim 8  wherein the  Lawsonia  spp. is  Lawsonia intracellularis.    
     
     
         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 contained within a deposited plasmid selected from the group consisting of AGAL Accession Nos: NM00/16476 (plasmid pGTE#1 glnH);    NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (plasmid pGTE#6 ntrC); NM00/16482 (plasmid pGTE#7 ytfM); and NM01/23286 (plasmid pGTE#8 ytfN).    
     
     
         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 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.  
     
     
         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  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.    
     
     
         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 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 AGAL Accession Nos: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (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 60% 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 60% 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: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (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: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (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;    (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 p plasmid selected from the group consisting of AGAL Accession Nos: NM00/16476 (plasmid pGTE#1 glnH); NM00/16477 (plasmid pGTE#2 flhB); NM00/16478 (plasmid pGTE#3 fliR); NM00/16479 (plasmid pGTE#4 motA/B); NM00/16480 (plasmid pGTE#5 tlyC); NM00/16481 (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.    
     
     
         21 . A combination vaccine composition for the prophylaxis or treatment of infection 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 of the  Lawsonia intracellularis  FIgE, 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.    
     
     
         22 - 50 . (canceled)