Salmonella vaccine materials and methods

Abstract

Attenuated mutant Salmonella bacteria containing inactivated virulence genes are provided for use in safe efficacious vaccines.

1 . (canceled)  
     
     
         2 . The vaccine composition of  claim 5  wherein said genes are inactivated by deletion of a portion of the coding region of the gene.  
     
     
         3 . The vaccine composition of  claim 5  wherein said genes are inactivated by an insertional mutation.  
     
     
         4 . (canceled)  
     
     
         5 . A vaccine composition comprising an immunologically protective amount of a first attenuated, non-reverting mutant  Salmonella  bacterium in which two or more secretion system apparatus (ssa) genes have been inactivated, wherein the genes are selected from the group consisting of ssaT, ssaJ, ssaC and ssaM, and wherein: 
 (a) said ssaT gene consists of SEQ ID NO: 1 or 2, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 1 or 2 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 1 or 2;    (b) said ssaJ gene consists of SEQ ID NO: 3 or 4, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 3 or 4 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 3 or 4;    (c) said ssaC gene consists of SEQ ID NO: 5 or 6, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 5 or 6 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 5 or 6; and    (d) said ssaM gene consists of SEQ ID NO: 7 or 30, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 7 or 30 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 7 or 30.    
     
     
         6 . (canceled)  
     
     
         7 . (canceled)  
     
     
         8 . The vaccine composition of  claim 5  further comprising a second attenuated mutant  Salmonella  bacterium in which one or more genes within the SPI2 region have been inactivated.  
     
     
         9 . The vaccine composition of  claim 8  wherein said genes within the SPI2 region are selected from the group consisting of ssaT, ssaJ, ssaC, and ssaM and wherein: 
 (a) said ssaT gene consists of SEQ ID NO: 1 or 2, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 1 or 2 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 1 or 2:    (b) said ssaJ gene consists of SEQ ID NO: 3 or 4, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 3 or 4 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 3 or 4;    (c) said ssaC gene consists of SEQ ID NO: 5 or 6, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 5 or 6 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 5 or 6; and    (d) said ssaM gene consists of SEQ ID NO: 7 or 30, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO: 7 or 30 under stringent conditions, or a full length  Salmonella  nucleotide sequence that has 95% sequence identity to SEQ ID NO: 7 or 30.    
     
     
         10 . The vaccine composition of  claim 8  wherein said first and second mutant  Salmonella  bacteria are from different serogroups.  
     
     
         11 . The vaccine composition of  claim 5  or  8  wherein said  Salmonella  bacteria are  Salmonella enterica  subsp  Enterica.    
     
     
         12 . The vaccine composition of  claim 5  or  8  wherein said  Salmonella  bacteria are from any of serogroups A, B, C1, C2, D1 or E1.  
     
     
         13 . The vaccine composition of  claim 8  wherein said first and second attenuated mutant  Salmonella  bacteria are selected from the group consisting of  S. dublin, S. choleraesuis  and  S. typhimurium.    
     
     
         14 . The vaccine composition of  claim 5  wherein said first attenuated mutant  Salmonella  bacterium further comprises a polynucleotide encoding a non- Salmonella  polypeptide.  
     
     
         15 . A method of conferring protective immunity on a non-rodent animal comprising the step of administering to said animal a vaccine composition comprising an immunologically protective amount of an attenuated, non-reverting mutant  Salmonella  bacterium according to  claim 5 .  
     
     
         16 . The method of  claim 15  wherein said immunologically protective amount of said attenuated bacterium provides an improvement in mortality, symptomatic diarrhea, physical condition, or milk production.  
     
     
         17 . The method of  claim 15  wherein said gene is inactivated by deletion of a portion of the coding region of the gene.  
     
     
         18 . (canceled)  
     
     
         19 . (canceled)  
     
     
         20 . A method of reducing the amount or duration of bacterial shedding during infection comprising the step of administering to a non-rodent animal a vaccine composition comprising an attenuated, non-reverting mutant  Salmonella  bacterium according to  claim 5 , in an amount effective to reduce bacterial shedding by said animal.  
     
     
         21 . The method of  claim 20  wherein the amount of bacterial shedding is reduced by about 10% or more.  
     
     
         22 . The method of  claim 20  wherein the duration of bacterial shedding is reduced by about 10% or more.  
     
     
         23 . (canceled)  
     
     
         24 . (canceled)  
     
     
         25 . The method of  claim 15  or  20  wherein said animal is selected from the group consisting of cattle, sheep, goats, horses, pigs, poultry and other birds, cats, dogs, and humans.  
     
     
         26 . The method of  claim 25  wherein said animal is a pig.  
     
     
         27 . The method of  claim 25  wherein said animal is cattle.  
     
     
         28 . A method of delivering a polypeptide antigen to an animal comprising the step of administering the vaccine composition of  claim 13  to said animal.  
     
     
         29 . The vaccine composition of  claim 5  wherein said genes are ssaJ and ssaC genes.  
     
     
         30 . The vaccine composition of  claim 5  wherein said genes are ssaM and ssaC genes.  
     
     
         31 . The vaccine composition of  claim 5  wherein said genes are ssaT and ssaC genes.  
     
     
         32 . The vaccine composition of  claim 5  wherein said genes are ssaT and ssaJ genes.  
     
     
         33 . The vaccine composition of  claim 5  wherein said gene is inactivated by deletion of the gene.  
     
     
         34 . The vaccine composition of  claim 5  wherein said genes are inactivated by a substitution within the coding region of the gene.  
     
     
         35 . The vaccine composition of  claim 5  wherein the first attenuated mutant  Salmonella  bacteria is selected from the group consisting of  S. dublin, S. choleraesuis  and  S. typhimurium.    
     
     
         36 . The vaccine composition of  claim 8 , wherein said first and second mutant  Salmonella  bacterium are from different serovars.  
     
     
         37 . The vaccine composition of  claim 8 , wherein each  Salmonella  bacterium is from any serovar of  S. bongoni  or  S. enterica.    
     
     
         38 . The method of  claim 15  wherein said gene is inactivated by deletion of the gene.  
     
     
         39 . The method of  claim 15  wherein said gene is inactivated by a substitution within the coding region of the gene.  
     
     
         40 . The method of  claim 25  wherein said bird is poultry.  
     
     
         41 . The method of  claim 25  wherein said animal is a horse.