Kim K J, McKinness S, Manclark C R
Laboratory of Pertussis, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892.
J Immunol. 1990 May 1;144(9):3529-34.
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.
为了解百日咳毒素分子的免疫学特性并探索开发合成疫苗的可能性,通过检测免疫小鼠淋巴结细胞和T细胞系对抗原及合成肽的增殖反应,对百日咳毒素酶促S1亚基上的T细胞表位进行了研究。用20 nM的酶促S1亚基刺激免疫淋巴细胞可获得最大的体外T细胞增殖反应。在检测具有不同MHC背景的小鼠淋巴细胞的T细胞增殖反应时,只有携带H-2d单倍型的小鼠对S1亚基是高反应者。为确定S1亚基上的T细胞表位,检测了BALB/c免疫淋巴细胞对几种合成S1肽的增殖反应。只有含有氨基酸残基65 - 79的肽被BALB/c淋巴细胞识别,并通过产生S1特异性BALB/c T细胞系证实其含有一个T细胞表位。利用该T细胞系,发现BALB/c小鼠对S1亚基以及对肽65 - 79的反应受限于II类抗原的I-Ad亚区。最后,我们发现用肽65 - 79免疫的小鼠的淋巴结细胞对天然S1亚基有反应。