Compositions relating to a mutant Clostridium difficile toxin and methods thereof

Abstract

In one aspect, the invention relates to an immunogenic composition that includes a mutant Clostridium difficile toxin A and/or a mutant Clostridium difficile toxin B. Each mutant toxin includes a glucosyltransferase domain having at least one mutation and a cysteine protease domain having at least one mutation, relative to the corresponding wild-type C. difficile toxin. The mutant toxins may further include at least one amino acid that is chemically crosslinked. In another aspect, the invention relates to antibodies or binding fragments thereof that binds to said immunogenic compositions. In further aspects, the invention relates to isolated nucleotide sequences that encode any of the foregoing, and methods of use of any of the foregoing compositions.

The invention claimed is: 
     
       1. A recombinant cell or progeny thereof expressing a polypeptide comprising SEQ ID NO: 84. 
     
     
       2. The A recombinant cell or progeny thereof according to  claim 1 , wherein the expressing a polypeptide comprises comprising the amino acid sequence SEQ ID NO: 4. 
     
     
       3. The recombinant cell or progeny thereof according to claim  1  2, comprising the nucleic acid sequence, which comprises SEQ ID NO: 12. 
     
     
       4. The recombinant cell or progeny thereof according to  claim 3 , comprising the nucleic acid sequence, which comprises SEQ ID NO: 45. 
     
     
       5. The recombinant cell or progeny thereof according to claim  1  2, wherein the cell is derived from a Gram positive bacterium cell. 
     
     
       6. The recombinant cell or progeny thereof according to claim  1  2, wherein the cell is a Clostridium difficile cell. 
     
     
       7. The recombinant cell or progeny thereof according to  claim 6 , further comprising an inactivated sporulation gene. 
     
     
       8. The recombinant cell or progeny thereof according to  claim 6 , wherein the cell lacks an endogenous polynucleotide encoding a Clostridium difficile toxin. 
     
     
       9. The recombinant cell or progeny thereof according to  claim 6 , wherein the cell is derived from the group consisting of Clostridium difficile 1351 (ATCC 43593™), C. difficile 3232 (ATCC BAA-1801™), C. difficile 7322 (ATCC 43601™), C. difficile 5036 (ATCC 43603™), C. difficile 4811 (ATCC 43602™), and C. difficile VPI 11186 (ATCC 700057™). 
     
     
       10. The recombinant cell or progeny thereof according to  claim 6 , wherein the cell is a Clostridium difficile VPI 11186 cell (ATCC 700057™). 
     
     
       11. The recombinant cell or progeny thereof according to claim  1  2, further comprising an additional recombinant nucleotide sequence. 
     
     
       12. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence encodes a mutant Clostridium difficile toxin. 
     
     
       13. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence encodes a mutant Clostridium difficile toxin A. 
     
     
       14. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence encodes a mutant Clostridium difficile toxin B. 
     
     
       15. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence encodes a polypeptide comprising SEQ ID NO: 86. 
     
     
       16. The recombinant cell or progeny thereof according to  claim 15 , wherein the polypeptide comprises SEQ ID NO: 6. 
     
     
       17. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence encodes a polypeptide comprising SEQ ID NO: 82. 
     
     
       18. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence encodes a polypeptide comprising SEQ ID NO: 85. 
     
     
       19. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence comprises SEQ ID NO: 14. 
     
     
       20. The recombinant cell or progeny thereof according to  claim 11 , wherein the recombinant nucleotide sequence comprises SEQ ID NO: 47. 
     
     
       21. A recombinant cell or progeny thereof expressing a polypeptide comprising SEQ ID NO 83.