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目的 研究胃癌组织中T淋巴细胞克隆的表型及其人白细胞抗原 (HLA)限制性。方法 通过有限稀释法建立T细胞克隆。应用免疫组织化学染色法检测T细胞克隆的表型 ,并以四甲基偶氮盐 (MTT)法检测T细胞克隆的细胞毒活性 ,然后应用HLA抗血清阻断抑制实验分析T细胞克隆的HLA限制性。采用放射免疫法检测T细胞克隆反应体系中肿瘤细胞坏死因子 β(TNF β)及γ 干扰素 (γ IFN)的释放。结果 共得到 3株可稳定增殖的T细胞克隆 ,分别标记为T1 0 8、T1 1 0、T30 2。其表型分别为 :CD3+ CD4 - CD8+ 、CD3+ CD4 + CD8- 及CD3+ CD4 + CD8- 。HLA抗血清阻断抑制实验表明T1 0 8HLA限制性为HLA A2。 3株T细胞克隆体系中TNF β的含量分别为 :1 6 .2 3、2 3 .67和 0 .34mg/L ;γ IFN的含量分别为 :2 0 .2 2、38.1 9和 2 7.64mg/L。结论 从胃癌组织中可分离得到T淋巴细胞克隆 ,T1 0 8的细胞毒活性受严格的HLA A2限制 ,T细胞克隆的细胞毒活性与其分泌的细胞因子有关
Objective To investigate the phenotype of T lymphocyte clones in gastric cancer and their human leucocyte antigen (HLA) restriction. Methods T cell clones were established by limiting dilution. The phenotypes of T cell clones were detected by immunohistochemical staining. The cytotoxic activity of T cell clones was detected by MTT assay, and then HLA anti - Restrictive. Radioimmunoassay was used to detect the release of tumor necrosis factor-β (TNF-β) and interferon-gamma (IFN-γ) in T cell cloning reaction system. Results A total of 3 stable T cell clones were obtained, which were labeled as T1 0 8, T1 1 0 and T30 2, respectively. The phenotypes were: CD3 + CD4 - CD8 +, CD3 + CD4 + CD8- and CD3 + CD4 + CD8-. HLA antisera blocking inhibition experiments showed that the restriction of T1 0 8 HLA was HLA A2. The content of TNFβ in the three T cell clones was respectively 16.22, 23.67 and 0.34 mg / L; the content of γ IFN was 20.22.38.1 9 and 2 7.64mg / L. Conclusion T lymphocyte clones can be isolated from gastric cancer tissue. The cytotoxic activity of T1 0 8 is restricted by the strict HLA A2. The cytotoxic activity of T cell clones is related to the secretion of cytokines