Immunological and biochemical characterization of streptococcal pyrogenic exotoxins I and J (SPE-I and SPE-J) from Streptococcus pyogenes.
Recently, we described the identification of novel streptococcal superantigens (SAgs) by mining the Streptococcus pyogenes M1 genome database at Oklahoma University. Here, we report the cloning, expression, and functional analysis of streptococcal pyrogenic exotoxin (SPE)-J and another novel SAg (SPE-I). SPE-I is most closely related to SPE-H and staphylococcal enterotoxin I, whereas SPE-J is most closely related to SPE-C. Recombinant forms of SPE-I and SPE-J were mitogenic for PBL, both reaching half maximum responses at 0.1 pg/ml. Evidence from binding studies and cell aggregation assays using a human B-lymphoblastoid cell line (LG-2) suggests that both toxins exclusively bind to the polymorphic MHC class II beta-chain in a zinc-dependent mode but not to the generic MHC class II alpha-chain. The results from analysis by light scattering indicate that SPE-J exists as a dimer in solution above concentrations of 4.0 mg/ml. Moreover, SPE-J induced a rapid homotypic aggregation of LG-2 cells, suggesting that this toxin might cross-link MHC class II molecules on the cell surface by building tetramers of the type HLA-DRbeta-SPE-J-SPE-J-HLA-DRbeta. SPE-I preferably stimulates T cells bearing the Vbeta18.1 TCR, which is not targeted by any other known SAG: SPE-J almost exclusively stimulates Vbeta2.1 T cells, a Vbeta that is targeted by several other streptococcal SAgs, suggesting a specific role for this T cell subpopulation in immune defense. Despite a primary sequence diversity of 51%, SPE-J is functionally indistinguishable from SPE-C and might play a role in streptococcal disease, which has previously been addressed to SPE-C.
Proft
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Division of Molecular Medicine and School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Arcus
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Handley
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Baker
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Fraser
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D019281:Dimerization; D005098:Exotoxins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000276:immunology; D005798:Genes, Bacterial; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008213:Lymphocyte Activation / Q000276:immunology; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D013297:Streptococcus pyogenes / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018089:Superantigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D015032:Zinc / Q000737:chemistry / Q000378:metabolism
D001426:Bacterial Proteins; D005098:Exotoxins; D000949:Histocompatibility Antigens Class II; D008024:Ligands; D008565:Membrane Proteins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins; C117980:SpeA protein, Streptococcus pyogenes; C423447:SpeJ protein, Streptococcus pyogenes; D018089:Superantigens; C027951:erythrogenic toxin; D015032:Zinc