Mycobacterial sulfation pathway proteins and methods of use thereof

Abstract

Novel mycobacterial sulfation pathway proteins and polypeptides related thereto, as well as nucleic acid compositions encoding the same, are provided. The subject polypeptide and nucleic acid compositions find use in a variety of applications, including research, diagnostic, and therapeutic agent screening applications. Also provided are methods of inhibiting growth and/or virulence of a pathogenic mycobacterium, and methods of treating disease conditions associated with a pathogenic mycobacterium, particularly by administering an inhibitor of a mycobacterial sulfation pathway protein. The present invention further provides genetically modified mycobacteria having a defect in a sulfation pathway enzyme gene; and immunogenic compositions that include such genetically modified mycobacteria.

1 . A genetically modified mycobacterium, wherein said genetically modified mycobacterium comprises a functionally disabled sulfation pathway gene, such that said sulfation pathway gene does not direct expression of a functional sulfation pathway polypeptide, wherein said genetically modified mycobacterium is avirulent. 
     
     
         2 . The mycobacterium of  claim 1 , wherein the mycobacterium has a functionally disabled APS reductase gene. 
     
     
         3 . The mycobacterium of  claim 1 , wherein the mycobacterium has a functionally disabled APS kinase gene. 
     
     
         4 . The mycobacterium of  claim 1 , wherein the mycobacterium has a functionally disabled sulfotransferase gene. 
     
     
         5 . The mycobacterium of  claim 1 , wherein the mycobacterium is  M. tuberculosis.    
     
     
         6 . The mycobacterium of  claim 1 , wherein the LD 50  is at least about 10-fold higher than a wild-type mycobacterium of the same strain. 
     
     
         7 . The mycobacterium of  claim 1 , wherein the LD 50  is at least about 50-fold higher than a wild-type mycobacterium of the same strain. 
     
     
         8 . An immunogenic composition comprising a genetically modified mycobacterium according to  claim 1 ; and a buffer. 
     
     
         9 . A pharmaceutical composition comprising a genetically modified mycobacterium according to  claim 1 ; and a pharmaceutically acceptable excipient. 
     
     
         10 . The composition of  claim 9 , further comprising an adjuvant. 
     
     
         11 . A kit comprising a sterile container comprising a genetically modified mycobacterium according to  claim 1 . 
     
     
         12 . The kit according to  claim 11 , wherein the sterile container comprises a unit dose of mycobacterium of from about 10 2  to about 10 7  mycobacterium. 
     
     
         13 . The kit according to  claim 11 , wherein the mycobacteria are lyophilized. 
     
     
         14 . The kit according to  claim 11 , wherein the sterile container further comprises a pharmaceutically acceptable excipient. 
     
     
         15 . The kit according to  claim 11 , wherein the sterile container further comprises an adjuvant. 
     
     
         16 . A method of increasing an immune response to a pathogenic mycobacterium in a host, comprising administering to the host an immunogenic composition according to  claim 8 . 
     
     
         17 . The method of  claim 16 , wherein said administering is intramuscular. 
     
     
         18 . The method of  claim 16 , wherein a protective immune response to a wild-type, virulent mycobacterium is induced. 
     
     
         19 . The method of  claim 18 , wherein the virulent mycobacterium is of the same species as the genetically modified mycobacterium. 
     
     
         20 . The method of  claim 18 , wherein the virulent mycobacterium is of a different species than the genetically modified mycobacterium. 
     
     
         21 . The method of  claim 16 , wherein cytotoxic T lymphocytes specific for mycobacteria are induced. 
     
     
         22 . A genetically modified mycobacterium, wherein said genetically modified mycobacterium comprises a modified sulfation pathway gene, such that said sulfation pathway gene does not direct expression of a sulfation pathway polypeptide, wherein said genetically modified mycobacterium is avirulent.