Identification of staphylococcal enterotoxin B sequences important for induction of lymphocyte proliferation by using synthetic peptide fragments of the toxin.
A series of 13 synthetic peptides, approximately 30 amino acids each, which spanned the entire sequence of staphylococcal enterotoxin B (SEB) were tested to evaluate their effects on T-cell proliferation in a culture system containing elutriated human peripheral blood lymphocytes incubated with a specific ratio of mononuclear cells. Four peptide regions were found to inhibit SEB-induced proliferation; they included sequences 1 to 30 (previously thought to be involved in major histocompatibility complex class II binding), 61 to 92 (sequences which relate to the T-cell receptor site), 93 to 112 (a linear sequence corresponding to the cysteine loop), and 130 to 160 (containing a highly conserved sequence, KKKVTAQEL). Antisera raised to this last peptide were capable of neutralizing SEB-induced proliferation. Antisera raised against the peptides which overlapped this sequence also were somewhat inhibitory. Neutralizing antisera were not produced from any other peptide sequence tested. To determine if any of these effects were nonspecific with regard to SEB-induced proliferation, the peptides were tested for inhibition of phorbol dibutyryl ester-induced proliferation, and only the sequence 93 to 112 (corresponding to the cysteinyl loop region) was consistently inhibitory (40%). Of the regions which displayed inhibition of SEB-induced proliferation, the peptide 130 to 160 inhibited binding of 125I-SEB to lymphocytes. These data suggest that the residues containing and surrounding the sequence KKKVTAQEL may be critical in the SEB-induced proliferation and may be useful for developing neutralizing antisera to SEB.
Jett
M
M
Division of Pathology, Walter Reed Army Institute of Research, Washington, D.C. 20307-5100.
Neill
R
R
Welch
C
C
Boyle
T
T
Bernton
E
E
Hoover
D
D
Lowell
G
G
Hunt
R E
RE
Chatterjee
S
S
Gemski
P
P
D000595:Amino Acid Sequence; D000818:Animals; D004768:Enterotoxins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology*; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D011817:Rabbits; D013211:Staphylococcus aureus / Q000472:pathogenicity*
D004768:Enterotoxins; D007106:Immune Sera; D010446:Peptide Fragments; D015240:Phorbol 12,13-Dibutyrate; C031818:enterotoxin B, staphylococcal