B cell epitope mapping of the bacterial superantigen staphylococcal enterotoxin B: the dominant epitope region recognized by intravenous IgG.
We showed that i.v. IgG contains Abs against a major group of bacterial superantigens, and that they can inhibit superantigen-elicited T cell activation. The B cell epitope region of the superantigen and the inhibitory mechanism have remained unknown. To analyze the dominant B cell epitopes on the bacterial superantigen SEB (staphylococcal enterotoxin B), we constructed fusion proteins of SEB deletion mutants, and the reactivities of these recombinant proteins to i.v. IgG and healthy human sera were evaluated by means of immunoblotting. Intravenous IgG and healthy human sera mostly recognized the C-terminal fragment (amino acid (aa) 133-239). The C-terminally truncated protein (aa 1-228) and the truncated mutant delta 225-234 lost reactivity, while the truncated protein (aa 1-234) did not, suggesting that the region (aa 225-234) is the dominant B cell epitope. The mutant, in which residues 226-229 of SEB were exchanged for residues 209-212 of streptococcal pyrogenic exotoxin A, reduced the reactivity with the C-terminal region-specific IgG purified by affinity chromatography. The C-terminal region-specific IgG inhibited SEB-elicited T cell activation, suggesting that this Ab that recognizes the epitope functions as the humoral defensive factor against SEB in humans. Furthermore, the assumed epitope region was homology to the residues (aa 32-41) of human thymopoietin, containing the biologic active site.
Nishi
J I
JI
Department of Pediatrics, Faculty of Medicine, Kagoshima University, Japan.
Kanekura
S
S
Takei
S
S
Kitajima
I
I
Nakajima
T
T
Wahid
M R
MR
Masuda
K
K
Yoshinaga
M
M
Maruyama
I
I
Miyata
K
K
D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000276:immunology*; D004768:Enterotoxins / Q000276:immunology*; D018604:Epitope Mapping; D006801:Humans; D016056:Immunodominant Epitopes / Q000032:analysis* / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D016756:Immunoglobulins, Intravenous / Q000032:analysis / Q000276:immunology*; D008969:Molecular Sequence Data; D013211:Staphylococcus aureus / Q000276:immunology*; D018089:Superantigens / Q000276:immunology*
D004768:Enterotoxins; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G; D016756:Immunoglobulins, Intravenous; D018089:Superantigens; C031818:enterotoxin B, staphylococcal