Determination of T cell epitopes on the S1 subunit of pertussis toxin.
To understand the immunologic characteristics of pertussis toxin molecule and to explore the possibility of developing a synthetic vaccine, T cell epitopes on the enzymatic S1 subunit of pertussis toxin were studied by measuring the proliferative response of immune murine lymph node cells and T cell lines to Ag and to synthetic peptides. The maximum in vitro T cell proliferative response was obtained by stimulating immune lymphoid cells with 20 nM of the enzymatic S1 subunit. When the T cell proliferative response of murine lymphoid cells with different MHC backgrounds was tested, only mice bearing the H-2d haplotype were high responder to the S1 subunit. To determine T cell epitopes on the S1 subunit, the proliferative response of BALB/c immune lymphoid cells to several synthetic S1 peptides was measured. Only the peptide containing amino acid residues, 65-79, was recognized by BALB/c lymphoid cells and was confirmed to contain a T cell epitope by generating S1 specific BALB/c T cell line. By using this T cell line, the response of BALB/c mice to the S1 subunit as well as to peptide 65-79 was shown to be restricted to the I-Ad sublocus of class II Ag. Finally, we showed that lymph node cells of mice immunized with peptide 65-79 respond to the native S1 subunit.
Laboratory of Pertussis, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892.
K J
KJ
Kim
S
S
McKinness
C R
CR
Manclark
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes; D066298:In Vitro Techniques; D008198:Lymph Nodes / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D037342:Pertussis Toxin; D013601:T-Lymphocytes / Q000276:immunology*; D010566:Virulence Factors, Bordetella / Q000276:immunology*
D000939:Epitopes; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin