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应用含重组OVA微小基因及全长基因的牛痘病毒感染能够稳定表达Kb 分子的多种真核细胞 ,通过构象特异的单克隆抗体 2 5 .D 1.16 (anti OVA2 57～ 2 64/Kb)检测细胞表面复合物的形成情况 ,并借助四聚体技术检测了它们对抗原特异性T细胞的诱导能力。结果表明四聚体的构建是成功的 ,进一步用已被重组牛痘病毒感染的L92 9 Kb 细胞作为抗原呈递细胞以诱导特异性细胞毒性T淋巴细胞 (CTL)的产生 ,并对活化后特异性CTL进行检测 ,从胞外IFN γ的分泌情况看 ,诱导后的整个群体细胞发生了活化与增殖 ,H 2Kb OVA四聚体对特异性T细胞进行的结合分析与经典的细胞毒效应相一致。可见 ,重组牛痘病毒介导的微小基因产物OVA2 57～ 2 64不仅能为MHC I类分子的合成提供更有效的肽来源 ,且能更有效诱导特异性CTL的产生。
The vaccinia virus containing recombinant OVA minigene and full-length gene was used to infect a variety of eukaryotic cells capable of stably expressing the Kb molecule. The cells were detected by conformation-specific monoclonal antibody 2.5D 1.16 (anti OVA2 57-264 / Kb) Surface complex formation and their ability to induce antigen-specific T cells was examined by means of tetramer technology. The results showed that the construction of tetramers was successful. L92 9 Kb cells infected with recombinant vaccinia virus were further used as antigen presenting cells to induce the production of specific cytotoxic T lymphocytes (CTLs), and the activation of specific CTLs For the detection of extracellular IFN-γ secretion, activation and proliferation of the entire population of cells after induction were observed. The binding analysis of H 2Kb OVA tetramers to specific T cells was consistent with classical cytotoxicity. It can be seen that OVA2 57-264 mediated by the recombinant vaccinia virus not only can provide a more effective peptide source for the synthesis of MHC class I molecules but also can induce the production of specific CTL more effectively.