Shimizu Y, Weidmann E, Iwatsuki S, Herberman R B, Whiteside T L
Pittsburgh Cancer Institute, Pennsylvania 15213.
Cancer Res. 1991 Nov 15;51(22):6153-62.
Three autotumor-reactive T-cell clones have been established from tumor-infiltrating lymphocytes isolated from a metastatic lesion of human gastric carcinoma in the liver. The clones all were shown to be CD3+, CD8+, CD4-, CD16-, T-cell receptor alpha/beta +, and T-cell receptor gamma/delta-, and they have retained both their autotumor reactivity and the same phenotype for over a year in culture. Each clone had a different rearrangement of the T-cell receptor gamma chain genes as indicated by Southern blot analysis. Tested against a panel of 18 tumor cell targets, the clones preferentially lysed autologous tumor (AuTu) cells, but each clone also showed weak cytotoxicity against one allogeneic cholangiocarcinoma cell line. At the same time, each clone showed appreciable cytotoxicity against K562 targets. In blocking experiments, anti-CD3, anti-WT31, anti-CD8, or anti-HLA class I monoclonal antibodies blocked AuTu cytotoxicity but not cytotoxicity against K562. In contrast, allocytotoxicity against the cholangiocarcinoma was blocked only by anti-CD3 monoclonal antibody. All 10 subclones of one T-cell clone had high levels of AuTu cytotoxicity but variable levels of anti-K562 cytotoxic activity. Proliferation of the T-cell clones was significantly stimulated by the addition of irradiated autologous but not allogeneic tumor cells. Preincubation of cultured AuTu cells with tumor necrosis factor alpha or gamma-interferon (IFN-gamma), but not with IFN-alpha, increased their susceptibility to lysis by the T-cell clones; however, it increased resistance of AuTu to lysis by interleukin 2-activated natural killer cells. The expression of an adhesion molecule, intercellular adhesion molecule 1, on the surface of AuTu was also up-regulated by tumor necrosis factor alpha or IFN-gamma, but not by IFN-alpha. All three cytokines up-regulated HLA-class-I antigens on AuTu. Pretreatment of K562 targets or allogeneic cholangiocarcinoma cells with the same cytokines increased their resistance to lysis by the T-cell clones. Overall, the results indicate that these T-cell clones show specificity for AuTu but also independently recognize a limited number of allogeneic tumor targets and lyse K562 targets. The mechanisms involved in the recognition by the T-cell clones of autologous, allogeneic, and K562 tumor targets appeared to be distinct.
从一名人类胃癌肝转移病灶中分离出的肿瘤浸润淋巴细胞中，已建立了三个自体肿瘤反应性T细胞克隆。这些克隆均显示为CD3 +、CD8 +、CD4 -、CD16 -、T细胞受体α/β +、T细胞受体γ/δ -，并且在培养一年多的时间里，它们既保留了自体肿瘤反应性，又保持了相同的表型。如Southern印迹分析所示，每个克隆的T细胞受体γ链基因都有不同的重排。在针对一组18种肿瘤细胞靶标的测试中，这些克隆优先裂解自体肿瘤（AuTu）细胞，但每个克隆对一种同种异体胆管癌细胞系也表现出较弱的细胞毒性。同时，每个克隆对K562靶标都表现出明显的细胞毒性。在阻断实验中，抗CD3、抗WT31、抗CD8或抗HLA I类单克隆抗体可阻断AuTu细胞毒性，但不能阻断对K562的细胞毒性。相比之下，对胆管癌的同种异体细胞毒性仅被抗CD3单克隆抗体阻断。一个T细胞克隆的所有10个子克隆都具有高水平的AuTu细胞毒性，但抗K562细胞毒性活性水平各不相同。添加经辐照的自体而非同种异体肿瘤细胞可显著刺激T细胞克隆的增殖。用肿瘤坏死因子α或γ干扰素（IFN-γ）而非IFN-α对培养的AuTu细胞进行预孵育，可增加它们对T细胞克隆裂解的敏感性；然而，这增加了AuTu对白细胞介素2激活的自然杀伤细胞裂解的抗性。肿瘤坏死因子α或IFN-γ可上调AuTu表面粘附分子细胞间粘附分子1的表达，但IFN-α不能。所有这三种细胞因子均可上调AuTu上的HLA I类抗原。用相同的细胞因子对K562靶标或同种异体胆管癌细胞进行预处理，可增加它们对T细胞克隆裂解的抗性。总体而言，结果表明这些T细胞克隆对AuTu具有特异性，但也能独立识别有限数量的同种异体肿瘤靶标并裂解K562靶标。T细胞克隆识别自体、同种异体和K562肿瘤靶标的机制似乎各不相同。
