GROUP A STREP IMMUNOGENIC COMPOSITIONS WITH POLYSACCHARIDE-PROTEIN CONJUGATES
The present disclosure provides immunogenic compositions comprising Group A Streptococcus (GAS) polypeptide antigens and at least one polypeptide-polysaccharide conjugate. The present disclosure further provides methods of using such compositions to induce immune responses against GAS infections in subjects.
1 . An immunogenic composition comprising
a. a first Group A Streptococcus (GAS) polypeptide antigen;
b. a second GAS polypeptide antigen; and
c. at least one polypeptide-polysaccharide conjugate, wherein the conjugate polypeptide is a third GAS polypeptide antigen or a non-GAS carrier polypeptide and comprises at least one non-natural amino acid, and wherein the conjugate polysaccharide is a GAS polysaccharide or a variant thereof that lacks an immunodominant N-acetyl Glucosamine (GlcNAc) side chain.
2 . The immunogenic composition of claim 1 , wherein the first and second GAS polypeptide antigens are independently selected from C5a peptidase, streptolysin O (SLO), Sib35, Sfb1, and SpyAD.
3 . The immunogenic composition of claim 1 , wherein the first and second GAS polypeptide antigens are C5a peptidase and SLO.
4 . The immunogenic composition of claim 1 , wherein the first and second GAS polypeptide antigens are C5a peptidase and SLO, and the C5a peptidase polypeptide antigen comprises the amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30, or a fragment thereof.
5 . (canceled)
6 . The immunogenic composition of claim 1 , wherein the SLO polypeptide antigen comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 31 or SEQ ID NO: 32.
7 . The immunogenic composition of claim 1 , wherein the first and second GAS polypeptide antigens are C5a peptidase and SLO, and the SLO polypeptide antigen comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, or SEQ ID NO: 53, or a fragment of any of the foregoing.
8 . The immunogenic composition of claim 1 , wherein the SLO polypeptide antigen comprises the amino acid sequence of SEQ ID NO: 31 or SEQ ID NO: 32, or a fragment thereof.
9 . (canceled)
10 . The immunogenic composition of claim 1 , wherein the first and second GAS polypeptide antigens are C5a peptidase and SLO, and the SLO polypeptide antigen comprises the amino acid sequence of SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, or SEQ ID NO: 53, or a fragment of any of the foregoing.
11 . (canceled)
12 . The immunogenic composition of claim 1 , wherein the first and second GAS polypeptide antigens are C5a peptidase and SpyAD.
13 . The immunogenic composition of claim 1 , wherein the C5a peptidase polypeptide antigen comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 29 or SEQ ID NO: 30, or a fragment thereof.
14 . The immunogenic composition of claim 1 , wherein the C5a peptidase polypeptide antigen comprises the amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30, or a fragment thereof.
15 - 16 . (canceled)
17 . The immunogenic composition of claim 1 , wherein the third GAS polypeptide antigen or non-GAS polypeptide antigen is selected from Streptococcus pyogenes Adhesion and Division (SpyAD) polypeptide, arginine deiminase (ADI), SEQ ID NO: 25, Protein D, and SLO, or a fragment of any of the foregoing.
18 . The immunogenic composition of claim 1 , wherein the nnAA comprises a click chemistry reactive group.
19 . The immunogenic composition of claim 1 , wherein the nnAA is selected from 2-amino-3-(4-azidophenyl)propanoic acid (pAF), 2-amino-4-azidobutanoic acid, 2-azido-3-phenylpropionic acid, 2-amino-3-azidopropanoic acid, 2-amino-3-(4-(azidomethyl)phenyl)propanoic acid (pAMF), 2-amino-3-(5-(azidomethyl)pyridin-2-yl)propanoic acid, 2-amino-3-(4-(azidomethyl)pyridin-2-yl)propanoic acid, 2-amino-3-(6-(azidomethyl)pyridin-3-yl)propanoic acid, and 2-amino-5-azidopentanoic acid.
20 . The immunogenic composition of claim 19 , wherein the nnAA is pAMF.
21 . The immunogenic composition of claim 17 , wherein the conjugate polypeptide is a third GAS protein, which is a SpyAD polypeptide and comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 33, or a fragment thereof.
22 . The immunogenic composition of claim 21 , wherein the SpyAD polypeptide comprises a pAMF substitution at positions K64, K287, K386, and K657 of SEQ ID NO: 33, or a fragment thereof.
23 . The immunogenic composition of claim 20 , wherein the SpyAD polypeptide comprises the amino acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34, or a fragment thereof.
24 . (canceled)
25 . The immunogenic composition of claim 17 , wherein the conjugate polypeptide is a third GAS polypeptide, which is an ADI polypeptide and comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 35 or SEQ ID NO: 36, or a fragment thereof.
26 . The immunogenic composition of claim 25 , wherein the ADI polypeptide comprises the amino acid sequence of SEQ ID NO: 27, SEQ ID NO: 37 or SEQ ID NO: 38, or a fragment thereof.
27 . (canceled)
28 . The immunogenic composition of claim 17 , wherein the conjugate polypeptide is a third GAS polypeptide, which is a SLO polypeptide and comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 31, SEQ ID NO: 32, or SEQ ID NO: 53, or a fragment of any of the foregoing.
29 . The immunogenic composition of claim 28 , wherein the SLO polypeptide comprises 3 or 4 pAMF substitutions at positions selected from K98, K112, R151, K189, K272, K323, K357, K375, K407, or K464.
30 . The immunogenic composition of claim 29 , wherein the SLO polypeptide comprises the amino acid sequence of SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, or SEQ ID NO: 64, or a fragment of any of the foregoing.
31 . (canceled)
32 . The immunogenic composition of claim 28 , wherein the SLO polypeptide comprises the amino acid sequence of SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, or SEQ ID NO: 76, or a fragment of any of the foregoing.
33 . (canceled)
34 . The immunogenic composition of claim 18 , wherein the conjugate polypeptide is a ferritin polypeptide and comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 21, or a fragment thereof.
35 . The immunogenic composition of claim 34 , wherein the ferritin polypeptide comprises a pAMF substitution at position 15 of SEQ ID NO: 21, or a fragment thereof.
36 . The immunogenic composition of claim 1 , wherein the conjugate polysaccharide is a GAS polysaccharide that lacks the immunodominant GlcNAc side chain.
37 . The immunogenic composition of claim 1 , wherein the conjugate polysaccharide has an average molecular weight of about 10 kDa to about 45 kDa.
38 . An immunogenic composition comprising
a. a Group A Streptococcus (GAS) C5a peptidase polypeptide antigen;
b. a GAS streptolysin O (SLO) polypeptide antigen; and
c. a polypeptide-polysaccharide conjugate comprising a Streptococcus pyogenes Adhesion and Division (SpyAD) conjugate polypeptide and a GAS conjugate polysaccharide or a variant thereof that lacks an immunodominant N-acetyl Glucosamine (GlcNAc) side chain.
39 . The immunogenic composition of claim 38 , wherein the SpyAD protein comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 33, or a fragment thereof, and comprises a pAMF substitution at positions K64, K287, K386, and K657.
40 . The immunogenic composition of claim 38 , wherein the SLO polypeptide antigen comprises the amino acid sequence of SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, or SEQ ID NO: 53, or a fragment of any of the foregoing.
41 . The immunogenic composition of claim 1 , comprising a C5a peptidase polypeptide antigen of SEQ ID NO: 30 or a fragment thereof; a SLO polypeptide antigen of SEQ ID NO: 53 or a fragment thereof; and a polypeptide-polysaccharide conjugate comprising a SpyAD conjugate polypeptide of SEQ ID NO: 34 or a fragment thereof and a GAS conjugate polysaccharide.
42 . The immunogenic composition of claim 1 , comprising a C5a peptidase polypeptide antigen of SEQ ID NO: 30 or a fragment thereof; a SLO polypeptide antigen of SEQ ID NO: 52 or a fragment thereof; and a polypeptide-polysaccharide conjugate comprising a SpyAD conjugate polypeptide of SEQ ID NO: 34 or a fragment thereof and a GAS conjugate polysaccharide.
43 . The immunogenic composition of claim 1 , comprising a C5a peptidase polypeptide antigen of SEQ ID NO: 30 or a fragment thereof; a SLO polypeptide antigen of SEQ ID NO: 32 or a fragment thereof; and a polypeptide-polysaccharide conjugate comprising a SpyAD conjugate polypeptide of SEQ ID NO: 34 or a fragment thereof and a GAS conjugate polysaccharide.
44 . The immunogenic composition of claim 1 , comprising a C5a peptidase polypeptide antigen of SEQ ID NO: 29 or a fragment thereof; a SLO polypeptide antigen of SEQ ID NO: 53 or a fragment thereof; and a polypeptide-polysaccharide conjugate comprising a SpyAD conjugate polypeptide of SEQ ID NO: 34 or a fragment thereof and a GAS conjugate polysaccharide.
45 . An immunogenic composition comprising
a. a Group A Streptococcus (GAS) C5a peptidase polypeptide antigen;
b. a GAS streptolysin O (SLO) polypeptide antigen; and
c. a polypeptide-polysaccharide conjugate comprising an arginine deiminase (ADI) conjugate polypeptide and a GAS conjugate polysaccharide or a variant thereof that lacks an immunodominant N-acetyl Glucosamine (GlcNAc) side chain.
46 . The immunogenic composition of claim 45 , wherein the ADI polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 35 or SEQ ID NO: 36, or a fragment thereof, and comprises a pAMF substitution at positions K15, K193, and K316.
47 . An immunogenic composition comprising
a. a Group A Streptococcus (GAS) C5a peptidase polypeptide antigen;
b. a GAS streptolysin O (SLO) polypeptide antigen; and
c. a polypeptide-polysaccharide conjugate comprising a ferritin polypeptide and a GAS conjugate polysaccharide or a variant thereof that lacks an immunodominant N-acetyl Glucosamine (GlcNAc) side chain.
48 . The immunogenic composition of claim 47 , wherein the ferritin protein comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 21 or a fragment thereof and comprises a pAMF substitution at position 15.
49 . The immunogenic composition of claim 1 , wherein the polypeptide-polysaccharide conjugate has an average molecular mass of about 10 kDa to about 5000 kDa.
50 . A method of inducing a protective immune response against a Group A Streptococcus (GAS) bacterium in a subject comprising administering the immunogenic composition of claim 1 to the subject.
51 . The method of claim 50 , wherein the immunogenic composition induces an antibody response in the subject against the Group A Streptococcus (GAS) bacterium and does not induce an antibody response in the subject against human tissue.
52 . The method of claim 50 , wherein the immunogenic composition induces a protective immune response against a Shigella bacterium in the subject, wherein the Shigella bacterium comprises a polysaccharide with a polyrhamnose backbone.
53 . A method of inducing a protective immune response against a Shigella bacterium in a subject comprising administering the immunogenic composition of claim 1 .
54 - 57 . (canceled)
58 . The immunogenic composition of claim 1 , wherein one or more of the first or second GAS polypeptide antigens is a fragment of the first or second polypeptide antigen.
59 . The immunogenic composition of claim 1 , wherein the third GAS polypeptide antigen is a fragment of the third GAS polypeptide antigen.