Patent
The present disclosure relates to immunogenic compositions for inducing an immune response in a subject for the treatment and/or prevention of a Staphylococcus aureus infection. The immunogenic compositions disclosed herein comprise a S. aureus protein A (SpA) polypeptide and a S. aureus Leukocidin A (LukA) and/or Leukocidin B (LukB) variant polypeptide. The present disclosure further relates to methods of generating an immune response against S. aureus in a subject that involve administering the disclosed immunogenic compositions.
1 . An immunogenic composition comprising: (i) a Staphylococcus aureus protein A (SpA) polypeptide, and (ii) a S. aureus LukA variant polypeptide, said LukA variant polypeptide comprising an amino acid substitution at each of amino acid residues corresponding to amino acid residues Lys83, Ser141, Val113, and Val193 of SEQ ID NO: 25.
2 . A combination of two or more immunogenic compositions, together comprising: (i) a Staphylococcus aureus protein A (SpA) polypeptide, and (ii) a S. aureus LukA variant polypeptide, said LukA variant polypeptide comprising an amino acid substitution at each of amino acid residues corresponding to amino acid residues Lys83, Ser141, Val113, and Val193 of SEQ ID NO: 25.
3 . The immunogenic composition of claim 1 , wherein the LukA variant polypeptide further comprises an amino acid substitution at the amino acid residue corresponding to Glu323 of SEQ ID NO: 25.
4 . The immunogenic composition of claim 3 , wherein the amino acid substitutions comprise Lys83Met, Ser141Ala, Val113Ile, Val193Ile, and Glu323 Ala.
5 . The immunogenic composition of claim 1 , wherein said LukA variant polypeptide comprises:
an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 3; or
an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 4; or
an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 5; or
an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 6; or
an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 7; or
an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 8.
6 . The immunogenic composition of claim 1 wherein said LukA variant polypeptide further comprises: an amino acid substitution at one or more amino acid residues corresponding to amino acid residues Tyr74, Asp140, Gly 149, and Gly156 of SEQ ID NO: 25.
7 . The immunogenic composition of claim 6 , wherein the amino acid substitutions comprise Tyr74Cys, Asp140Cys, Gly 149Cys, and Gly156Cys.
8 . The immunogenic composition of claim 1 , wherein said variant LukA protein or polypeptide further comprises: an amino acid substitution at the amino acid residue corresponding to amino acid residue Thr249 of SEQ ID NO: 25.
9 . The immunogenic composition of claim 1 , wherein the SpA polypeptide is a SpA variant polypeptide.
10 . The immunogenic composition of claim 9 , wherein the SpA variant polypeptide has at least one amino acid substitution that disrupts Fc binding and at least a second amino acid substitution that disrupts VH3 binding.
11 . The immunogenic composition of claim 9 , wherein the SpA variant polypeptide comprises a SpA D domain, said SpA D domain comprising an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 58.
12 . The immunogenic composition of claim 10 , wherein the SpA variant polypeptide has an amino acid substitution at one or both of amino acid positions corresponding to positions 9 and 10 of SEQ ID NO: 58.
13 . The immunogenic composition of claim 12 , wherein the SpA variant polypeptide further comprises a SpA E, A, B, or C domain, wherein the SpA variant polypeptide has an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 54, and/or wherein each SpA E, A, B, and C domain has an amino acid substitution at one or both amino acid positions corresponding to amino acid positions 9 and 10 of SEQ ID NO: 58.
14 . The immunogenic composition of claim 12 wherein the amino acid substitution at one or both amino acid positions corresponding to positions 9 and 10 of SEQ ID NO: 58 is a lysine residue for a glutamine residue.
15 . The immunogenic composition of claim 9 , wherein the SpA variant polypeptide comprises at least one SpA A, B, C, D, or E domain, and wherein the at least one domain has (i) a lysine substitution at the glutamine residues corresponding to positions 9 and 10 of SEQ ID NO: 58 and (ii) a glutamate substitution at the amino acid position corresponding to position 33 of SEQ ID NO: 58.
16 . The immunogenic composition of claim 1 , wherein said composition further comprises a S. aureus Leukocidin B (LukB) polypeptide or variant thereof.
17 . The immunogenic composition of claim 16 , wherein the LukB polypeptide is a LukB polypeptide of SEQ ID NO: 15 or a LukB polypeptide of SEQ ID NO: 16.
18 . The immunogenic composition of claim 16 , wherein the LukB polypeptide is a LukB variant polypeptide.
19 . The immunogenic composition of claim 18 , wherein the LukB variant polypeptide comprises an amino acid sequence having at least 85% sequence similarity to the amino acid sequence of SEQ ID NO:15 or an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 16.
20 . The immunogenic composition of claim 19 , wherein the LukB variant polypeptide comprises an amino acid substitution at the amino acid position corresponding to position 53 of SEQ ID NO: 15 and SEQ ID NO: 16.
21 . The immunogenic composition of claim 20 , wherein the amino acid substitution is a valine to leucine substitution.
22 . The immunogenic composition of claim 18 , wherein said LukB variant polypeptide comprises an amino acid substitution at one or more amino acid residues corresponding to amino acid residues Glu45, Glu109, Thr121, and Arg154 of SEQ ID NO: 15, and/or wherein said LukB variant polypeptide comprises an amino acid substitution at one or more amino acid residues corresponding to amino acid residues Glu45, Glu110, Thr122, and Arg155 of SEQ ID NO: 16.
23 . The immunogenic composition of claim 18 , wherein the LukB variant polypeptide comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 17-22.
24 . The immunogenic composition of claim 16 , wherein said composition comprises a LukA variant polypeptide comprising the amino acid sequence SEQ ID NO: 4 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 4 and a LukB polypeptide comprising the amino acid sequence of SEQ ID NO: 16, or an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 16; or
wherein said composition comprises a LukA variant polypeptide comprising the amino acid sequence SEQ ID NO: 3 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 3 and a LukB polypeptide comprising the amino acid sequence of SEQ ID NO: 15, or an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 15; or
wherein said composition comprises a LukA variant polypeptide comprising the amino acid sequence SEQ ID NO: 3 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 3 and a LukB polypeptide comprising the amino acid sequence of SEQ ID NO: 18, or an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18.
25 . The immunogenic composition of claim 24 , wherein the SpA polypeptide is a SpA variant polypeptide.
26 . The immunogenic composition of claim 25 , wherein the SpA variant polypeptide comprises at least one SpA A, B, C, D, or E domain, and wherein the at least one domain has (i) a lysine substitution at the glutamine residues corresponding to positions 9 and 10 of SEQ ID NO: 58 and (ii) a glutamate substitution at the amino acid position corresponding to position 33 of SEQ ID NO: 58.
27 . The immunogenic composition of claim 16 , wherein (i) the SpA variant polypeptide comprises at least one SpA A, B, C, D, or E domain, and wherein the at least one domain has lysine substitutions at the amino acid positions corresponding to positions 9 and 10 of SEQ ID NO: 58 and a glutamate substitution at the amino acid position corresponding to position 33 of SEQ ID NO: 58; (ii) the LukA variant polypeptide comprises a CC8 LukA variant polypeptide comprising a methionine substitution at the amino acid position corresponding to position 80 of SEQ ID NO: 1, an alanine substitution at the amino acid position corresponding to position 138 of SEQ ID NO: 1, an isoleucine substitutions at the amino acid positions corresponding to positions 110 and 190 of SEQ ID NO:1, and an alanine substitution at the amino acid position corresponding to position 320 of SEQ ID NO: 1; and (iii) the LukB polypeptide is a CC45 LukB variant polypeptide comprising a leucine substitution at the amino acid position corresponding to position 53 of SEQ ID NO: 16.
28 . The immunogenic composition of claim 27 , wherein the SpA variant polypeptide comprises consecutively SpA E, D, A, B, and C domains, each domain having the lysine substitutions at the amino acid positions corresponding to positions 9 and 10 of SEQ ID NO: 58 and a glutamate substitution at the amino acid position corresponding to position 33 of SEQ ID NO: 58.
29 . The immunogenic composition of claim 1 , further comprising an adjuvant.
30 . The immunogenic composition of claim 29 , wherein the adjuvant comprises a stable oil-in-water emulsion, a saponin, a TLR4 agonist, or a combination thereof.
31 . The immunogenic composition of claim 30 , wherein the adjuvant comprises a saponin and the saponin is QS21.
32 . The immunogenic composition of claim 30 , wherein the adjuvant comprises a TLR4 agonist and the TLR4 agonist is lipid A or an analog or derivative thereof, and/or wherein the TLR4 agonist is glycopyranosyl lipid adjuvant (GLA).
33 . The immunogenic composition of claim 32 , wherein the TLR4 agonist is GLA-SE or GLA-LSQ.
34 . The immunogenic composition of claim 30 , wherein the adjuvant comprises a TLR-4 agonist in combination with either a saponin or a stable oil-in-water emulsion.
35 . An immunogenic composition, wherein said composition comprises one or more nucleic acid molecules encoding the Staphylococcus aureus protein A (SpA) polypeptide, the LukA variant polypeptide, and the LukB polypeptide or variant thereof of the immunogenic compositions of claim 16 .
36 . The immunogenic composition of claim 35 , wherein the one or more nucleic acid molecules are contained in one or more vectors.
37 . The immunogenic composition of claim 36 , wherein said composition comprises a host cell, wherein said host cell comprises said one or more nucleic acid molecules or said one or more vectors.
38 . A method for treating or preventing a Staphylococcus infection in a subject in need thereof, the method comprising: administering to the subject in need thereof an effective amount of the immunogenic composition of claim 1 .
39 . A method for eliciting an immune response to a Staphylococcus bacterium in a subject in need thereof, the method comprising: administering to the subject in need thereof an effective amount of the immunogenic composition of claim 1 .
40 . A method for decolonization or preventing colonization or recolonization of a Staphylococcus bacterium in a subject in need thereof, the method comprising: administering to the subject in need thereof an effective amount of the immunogenic composition of claim 1 .