胃癌组织中T淋巴细胞克隆的研究

Study of T-cell clones from the tumor tissue of a patient with gastric carcinoma

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 。

Objectives To study the phenotype and human leukocyte antigen (HLA) restriction of T cell clones established from the tumor tissue in a patient with gastric carcinoma.Methods T cell clones were isolated by limiting dilution.T cell clones' phenotype was analyzed by immunohistochemistry and the cytotoxic activities of T cell clones detected by MTT test.Analysis of HLA restriction were carried out by HLA anti serum inhibition assay.TNF β and γ IFN production were examined by RIA. Results The phenotypes of three strains of T cell clones,labelled with T108,T110 and T302,were CD 3 +CD 4 -CD 8 +,CD 3 +CD 4 +CD 8 -and CD 3 +CD 4 +CD 8 -,respectively.HLA anti serum blocking inhibitory test indicated that HLA constriction of T108 was HLA A2.The average contents of TNF β in the T cell clone system of three strains were 16.23,23.67 and 0.34 mg/L respectively and those of γ IFN 20.22,38.19 and 27.64 mg/L respectively.Conclusion It was able to separate TIL from gastric carcinoma patients.The cytotoxic activity of T108 was strictly restricted by HLA A2.The cytotoxic activity of T lymphocytes might be related with the secreted cytokines.