Celis E, Ou D, Dietzschold B, Otvos L, Koprowski H
Wistar Institute of Anatomy and Biology, Philadelphia, PA.
Hybridoma. 1989 Jun;8(3):263-75. doi: 10.1089/hyb.1989.8.263.
Rabies virus-specific T-cell hybridomas were produced from immune mice by somatic cell fusion. Cloned T-cell hybridomas were studied for antigen specificity using purified virus, a recombinant vaccinia virus expressing rabies glycoprotein and synthetic peptides containing amino acid sequences of rabies viral antigens. Two closely situated T-cell epitopes of rabies glycoprotein, and one of rabies nucleoprotein were identified using synthetic peptides corresponding to amino acid sequences of these proteins. The major histocompatibility gene complex elements that determine the recognition of antigen by these T-cell hybrids were determined using mouse fibroblasts (L cells) transfected with and expressing the I-Ad and I-Ed genes. Some of the T cell hybridomas exhibited significant cytotoxic activity against target cells expressing surface rabies antigens. This T cell mediated cytotoxicity requires cell-to-cell contact between target and effector cells since no by-stander cytotoxicity was observed. The results are discussed in the context of their significance for the design of newer subunit vaccines to prevent rabies infection.
通过体细胞融合从免疫小鼠中产生了狂犬病病毒特异性T细胞杂交瘤。使用纯化病毒、表达狂犬病糖蛋白的重组痘苗病毒以及含有狂犬病病毒抗原氨基酸序列的合成肽，对克隆的T细胞杂交瘤进行抗原特异性研究。使用与这些蛋白质氨基酸序列相对应的合成肽，鉴定出狂犬病糖蛋白的两个紧密相邻的T细胞表位以及狂犬病核蛋白的一个T细胞表位。使用转染并表达I-Ad和I-Ed基因的小鼠成纤维细胞（L细胞），确定了决定这些T细胞杂交瘤对抗原识别的主要组织相容性基因复合体元件。一些T细胞杂交瘤对表达表面狂犬病抗原的靶细胞表现出显著的细胞毒性活性。这种T细胞介导的细胞毒性需要靶细胞和效应细胞之间的细胞间接触，因为未观察到旁观者细胞毒性。结合这些结果对于设计预防狂犬病感染的新型亚单位疫苗的意义进行了讨论。
