Multivalent live vector vaccine against
The invention relates to a multivalent Clostridium difficile vaccine comprising a Salmonella Typhi live vector comprising the cell binding domain of TcdA toxin (CBD/A) of Clostridium difficile or an antigenic fragment thereof and the cell binding domain of TcdB toxin (CBD/B) of Clostridium difficile or an antigenic fragment thereof and optionally the cell-binding subunit component (CdtB) of binary toxin of Clostridium difficile or an antigenic fragment thereof. The invention further provides methods of inducing an immune response and methods of preventing recurrence of C. difficile infections in subjects.
1. An immunogenic composition comprising a live Salmonella Typhi vector comprising a cell binding domain of TcdA toxin (CBD/A) of Clostridium difficile , a cell binding domain of TcdB toxin (CBD/B) of Clostridium difficile and a cell-binding subunit (CdtB) of the Clostridium difficile colonization factor binary toxin, wherein at least one of said toxins is inserted into a Salmonella Typhi chromosomal locus selected from the group consisting of guaBA and htrA.
2. The immunogenic composition of claim 1 , wherein the TcdB toxin (CBD/B) is expressed on a plasmid in Salmonella Typhi.
3. The immunogenic composition of claim 2 , wherein the plasmid has a non-antibiotic based plasmid selection system.
4. The immunogenic composition of claim 3 , wherein the plasmid expresses a gene that is essential for the growth of Salmonella Typhi and has been chromosomally mutated in Salmonella Typhi.
5. The immunogenic composition of claim 4 , wherein the gene encodes single stranded binding protein (SSB).
6. The immunogenic composition of claim 5 , wherein the TcdA toxin (CBD/A) and CdtB from binary toxin are chromosomally integrated in Salmonella Typhi.
7. The immunogenic composition of claim 6 , wherein the TcdA toxin (CBD/A), the TcdB toxin (CBD/B) and CdtB from binary toxin are fused to cytolysin A (ClyA) protein from Salmonella Typhi to facilitate export from the cell.
8. The immunogenic composition of claim 7 , wherein ClyA has been mutated to reduce hemolytic activity of ClyA and the ClyA mutant is selected from the group consisting of ClyA I198N and ClyA C285W, wherein the isoleucine residue at position 198 and the cysteine residue at position 285 correspond to isoleucine and cysteine residues at those positions in SEQ ID NO: 14.
9. The immunogenic composition of claim 6 , wherein the TcdA toxin (CBD/A) and CdtB from binary toxin are inserted into the guaBA locus of Salmonella Typhi.
10. The immunogenic composition of claim 6 , wherein CdtB from binary toxin is inserted into the htrA locus of Salmonella Typhi and the TcdA toxin (CBD/A) is inserted into the guaBA locus of Salmonella Typhi.
11. The immunogenic composition of claim 6 , wherein one or more nucleic acid sequences encoding the TcdA toxin (CBD/A), the TcdB toxin (CBD/B) or CdtB from binary toxin are codon optimized for expression in Salmonella Typhi.