Vaccine against necrotic enteritis in poultry

Abstract

An immunogenic polypeptide selected from an isolated Clostridium perfringens pilus polypeptide, a variant of the pilus polypeptide; a fragment of the pilus polypeptide; and a fragment of the variant, is useful for the preparation of a vaccine for the treatment or prevention of enteric necrosis in poultry. The isolated Clostridium perfringens pilus polypeptide includes an assembled pilus or the pilus subunits CnaA, FimA and/or FimB.

1 . A method of treating or preventing necrotic enteritis or  Clostridium perfringens  infection in poultry, the method comprising administering to the poultry an effective amount of an immunogenic polypeptide or an effective amount of a vaccine comprising the immunogenic polypeptide,
 wherein the immunogenic polypeptide is selected from the group consisting of an isolated  Clostridium perfringens  pilus polypeptide, a variant of the pilus polypeptide, a fragment of the pilus polypeptide, and a fragment of the variant of the pilus polypeptide;   wherein the pilus polypeptide, the variant of the pilus polypeptide, the fragment of the pilus polypeptide and the fragment of the variant of the pilus polypeptide are immunogenic in poultry;   wherein the pilus polypeptide is an assembled sortase-dependent pilus or a subunit thereof, and   wherein the pilus polypeptide is encoded by one or more genes in the VR-10B genetic locus.   
     
     
         2 . The method according to  claim 1  wherein the variant of the pilus polypeptide has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99% or at least 99.9% sequence identity to the pilus polypeptide. 
     
     
         3 . The method according to  claim 1 , wherein the subunit of the assembled sortase-dependent pilus is selected from the group consisting of a CnaA polypeptide, a FimA polypeptide, and a FimB polypeptide; and wherein the assembled sortase-dependent pilus comprises one or more subunits each individually selected from the CnaA polypeptide, the FimA polypeptide, and the FimB polypeptide. 
     
     
         4 . The method according to  claim 3 , wherein the CnaA polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:10 and SEQ ID NO:13. 
     
     
         5 . The method according to  claim 3 , wherein the CnaA polypeptide is encoded by a polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4 and SEQ ID NO:7. 
     
     
         6 . The method according to  claim 3 , wherein the CnaA polypeptide is encoded by a polynucleotide which hybridizes under at least moderately stringent conditions to a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4 and SEQ ID NO:7. 
     
     
         7 . The method according to  claim 3 , wherein the FimA polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:11 and SEQ ID NO:14. 
     
     
         8 . The method according to  claim 3 , wherein the FimA polypeptide is encoded by a polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5 and SEQ ID NO:8. 
     
     
         9 . The method according to  claim 3 , wherein the FimA polypeptide is encoded by a polynucleotide which hybridizes under at least moderately stringent conditions to a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5 and SEQ ID NO:8. 
     
     
         10 . The method according to  claim 3 , wherein the FimB polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:12 and SEQ ID NO:15. 
     
     
         11 . The method according to  claim 3 , wherein the FimB polypeptide is encoded by a polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6 and SEQ ID NO:9. 
     
     
         12 . The method according to  claim 3 , wherein the FimB polypeptide is encoded by a polynucleotide which hybridizes under at least moderately stringent conditions to a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6 and SEQ ID NO:9. 
     
     
         13 . A vaccine comprising an effective amount of an immunogenic polypeptide and an adjuvant,
 wherein the immunogenic polypeptide is selected from the group consisting of an isolated  Clostridium perfringens  pilus polypeptide, a variant of the pilus polypeptide, a fragment of the pilus polypeptide, and a fragment of the variant of the pilus polypeptide;   wherein the pilus polypeptide, the variant of the pilus polypeptide, the fragment of the pilus polypeptide and the fragment of the variant of the pilus polypeptide are immunogenic in poultry;   wherein the pilus polypeptide is an assembled sortase-dependent pilus or a subunit thereof,   wherein the pilus polypeptide is encoded by one or more genes in the VR-10B genetic locus, and   wherein the vaccine is effective to treat or prevent necrotic enteritis or  Clostridium perfringens  infection in poultry.   
     
     
         14 . The vaccine according to  claim 13  wherein the variant of the pilus polypeptide has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99% or at least 99.9% sequence identity to the pilus polypeptide. 
     
     
         15 . The vaccine according to  claim 13 , wherein the subunit of the assembled sortase-dependent pilus is selected from the group consisting of a CnaA polypeptide, a FimA polypeptide, and a FimB polypeptide; and wherein the assembled sortase-dependent pilus comprises one or more subunits each individually selected from the CnaA polypeptide, the FimA polypeptide, and the FimB polypeptide. 
     
     
         16 . The vaccine according to  claim 15 , wherein the CnaA polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:10 and SEQ ID NO:13. 
     
     
         17 . The vaccine according to  claim 15 , wherein the CnaA polypeptide is encoded by a polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4 and SEQ ID NO:7. 
     
     
         18 . The vaccine according to  claim 15 , wherein the CnaA polypeptide is encoded by a polynucleotide which hybridizes under at least moderately stringent conditions to a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4 and SEQ ID NO:7. 
     
     
         19 . The vaccine according to  claim 15 , wherein the FimA polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:11 and SEQ ID NO:14. 
     
     
         20 . The vaccine according to  claim 15 , wherein the FimA polypeptide is encoded by a polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5 and SEQ ID NO:8. 
     
     
         21 . The vaccine according to  claim 15 , wherein the FimA polypeptide is encoded by a polynucleotide which hybridizes under at least moderately stringent conditions to a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5 and SEQ ID NO:8. 
     
     
         22 . The vaccine according to  claim 15 , wherein the FimB polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:12 and SEQ ID NO:15. 
     
     
         23 . The vaccine according to  claim 15 , wherein the FimB polypeptide is encoded by a polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6 and SEQ ID NO:9. 
     
     
         24 . The vaccine according to  claim 15 , wherein the FimB polypeptide is encoded by a polynucleotide which hybridizes under at least moderately stringent conditions to a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6 and SEQ ID NO:9. 
     
     
         25 . A method of treating or preventing necrotic enteritis or  Clostridium perfringens  infection in poultry, the method comprising administering to the poultry an effective amount of a vaccine according to  claim 13 . 
     
     
         26 . A method of immunizing a subject, the method comprising administering an effective amount of a vaccine according to  claim 13  to a subject. 
     
     
         27 . The method of  claim 26  wherein the subject is poultry.