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目的 体外建立可长期培养的抗原特异性T细胞系及克隆 ,并分析其T细胞受体的使用情况。方法 用卵清白蛋白 (OVA)免疫BALB c小鼠 ,取淋巴结细胞在体外用抗原反复刺激建立抗原特异性T细胞系 ,通过有限稀释法进行克隆 ,应用锚定PCR方法分析其特异的T细胞受体的应用。结果 建立了H 2 d 限制的OVA特异性T细胞系 ,获得了 3株特异性较高的T细胞克隆 ,T细胞系中TCRAV11S4和BV4S1应用频率最高 ,3株T细胞克隆的TCR均为AV5S2和BV2S1,这些T细胞所使用的TCRα链和 β链在CDR3区有明显的共性。结论 获得了卵清白蛋白特异的T细胞系和克隆 ,且其T细胞受体的应用有一定的限制性
Objective To establish a long-term culture of antigen-specific T cell lines and clones in vitro and analyze their T cell receptor usage. Methods BALBc mice were immunized with ovalbumin (OVA). Antigen-specific T cell lines were established by repeatedly stimulating lymph node cells with antigen in vitro and cloning by limiting dilution. The specific T cell receptor Body application. Results The H 2 d-restricted OVA-specific T cell line was established and three highly specific T cell clones were obtained. TCRAV11S4 and BV4S1 were the most frequently used T cell lines. The TCRs of the three T cell clones were both AV5S2 and BV2S1, the TCRα chain and the β chain used by these T cells have obvious commonalities in the CDR3 region. Conclusion Ovalbumin-specific T cell lines and clones were obtained, and the application of T cell receptors was limited