Immunogenic compositions comprising
The present disclosure provides immunogenic compositions useful in prevention and treatment of Staphylococcus aureus infection. In particular, the disclosure provides methods of inducing an immune response against a LukAB-expressing S. aureus, methods of preventing or treating S. aureus infections, and composition for preventing or treating S. aureus infections.
1. An isolated mutant staphylococcal leukocidin subunit polypeptide comprising a wild-type staphylococcal LukA subunit, a wild-type staphylococcal LukB subunit, or a wild-type staphylococcal LukAB dimer, except for having one or more amino acid substitutions at conserved residues in the LukA subunit, the LukB subunit, or in the LukAB dimer,
comprising a mutation in the LukAB dimer/dimer interface region,
wherein the mutation in the LukAB dimer/dimer interface region is at a LukB position corresponding to R23 of SEQ ID NO: 29, and
wherein the LukB position corresponding to R23 of SEQ ID NO: 29 is substituted with glutamate (E).
2. The mutant subunit polypeptide of claim 1 , further comprising a mutation at a LukA position corresponding to D 39 of SEQ ID NO: 28, a LukA position corresponding to D75 of SEQ ID NO: 28, a LukA position corresponding to K138 of SEQ ID NO: 28, a LukA position corresponding to D197 of SEQ ID NO: 28, a LukB position corresponding to K12 of SEQ ID NO: 29, a LukB position corresponding to K19 of SEQ ID NO: 29, a LukB position corresponding to K58 of SEQ ID NO: 29, a LukB position corresponding to E112 of SEQ ID NO: 29, a LukB position corresponding to K218 of SEQ ID NO: 29, or any combination thereof.
3. The mutant subunit polypeptide of claim 1 , further comprising a mutation at a LukA position corresponding to D39 of SEQ ID NO: 28,
wherein the LukA position corresponding to D39 of SEQ ID NO: 28 is substituted with alanine (A) or arginine (R).
4. The mutant subunit polypeptide of claim 1 , further comprising a mutation at a LukA position corresponding to D39 of SEQ ID NO: 28,
wherein the LukA position corresponding to D39 of SEQ ID NO: 28 is substituted with alanine (A).
5. The mutant subunit polypeptide of claim 2 , wherein the LukA position corresponding to D75 of SEQ ID NO: 28 is substituted with alanine (A).
6. The mutant subunit polypeptide of claim 2 , wherein the LukA position corresponding to K138 of SEQ ID NO: 28 is substituted with alanine (A).
7. The mutant subunit polypeptide of claim 2 , wherein the LukA position corresponding to D197 of SEQ ID NO: 28 is substituted with alanine (A) or lysine (K).
8. The mutant subunit polypeptide of claim 2 , wherein the LukB position corresponding to K12 of SEQ ID NO: 29 is substituted with alanine (A) and/or the LukB position corresponding to K19 of SEQ ID NO: 29 is substituted with alanine (A).
9. The mutant subunit polypeptide of claim 2 , comprising a triple-mutant LukB, wherein the LukB position corresponding to K12 of SEQ ID NO: 29 is substituted with alanine (A), the LukB position corresponding to K19 of SEQ ID NO: 29 is substituted with alanine (A), and the LukB position corresponding to R23 of SEQ ID NO: 29 is substituted with glutamate (E).
10. The mutant subunit polypeptide of claim 2 , wherein the LukB position corresponding to K58 of SEQ ID NO: 29 is substituted with alanine (A) or glutamate (E).
11. The mutant subunit polypeptide of claim 2 , wherein the LukB position corresponding to E112 of SEQ ID NO: 29 is substituted with alanine (A).
12. The mutant subunit polypeptide of claim 2 , wherein the LukB position corresponding to K218 of SEQ ID NO: 29 is substituted with alanine (A).
13. The mutant subunit polypeptide of claim 1 , which is less toxic in a neutrophil toxicity assay compared to the corresponding wild-type leukocidin subunit.
14. The mutant subunit polypeptide of claim 2 , wherein the wild-type LukA subunit comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 35, and SEQ ID NO: 41.
15. The mutant subunit polypeptide of claim 1 , wherein the wild-type LukB subunit comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 17, SEQ ID NO: 49, SEQ ID NO: 53, and SEQ ID NO: 54.
16. A polypeptide complex comprising a mutant LukAB dimer of claim 1 ,
wherein the mutant LukAB dimer comprises both a mutant LukA subunit and a mutant LukB subunit, and
wherein the LukAB dimer comprises LukB R23E.
17. The polypeptide complex of claim 16 , wherein the LukAB dimer comprises LukA D39A or LukA D39R.
18. The polypeptide complex of claim 17 comprising LukA D39A and LukB R23E.
19. A composition comprising the mutant subunit polypeptide of claim 3 , and a carrier.
20. The mutant subunit polypeptide of claim 2 , wherein the wild-type LukB subunit comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 17, SEQ ID NO: 49, SEQ ID NO: 53, and SEQ ID NO: 54.
21. The mutant subunit polypeptide of claim 2 , wherein the wild-type LukA subunit comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 35, and SEQ ID NO: 41, and wherein the wild-type LukB subunit comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 17, SEQ ID NO: 49, SEQ ID NO: 53, and SEQ ID NO: 54.