de Vries J E, Spits H
J Immunol. 1984 Jan;132(1):510-9.
Activation of peripheral blood lymphocytes (PBL) from a melanoma patient either in secondary MLC in which EBV-transformed B cells from the cell line JY were used as stimulator cells, or by co-cultivation with the autologous melanoma cells in a mixed leukocyte tumor cell culture (MLTC) resulted in the generation of cytotoxic activity against the autologous melanoma (O-mel) cells. From these activated bulk cultures four cloned cytotoxic T lymphocyte (CTL) lines were isolated. The CTL clone O-1 (T3+, T4+, T8-, OKM-1-, HNK-, and HLA-DR+), and O-36 (T3+, T4-, T8+, OKM-, HNK-, and HLA-DR+) were obtained from MLC, whereas the CTLC clones O-C7 (T3+, T4+, T8-, OKM-1-, HNK-, and HLA-DR+) and O-D5 (T3+, T4-, T8+, OKM-1-, HNK, and HLA-DR+) were isolated from autologous MLTC. All four CTL clones were strongly cytotoxic for O-mel cells but failed to lyse autologous fibroblasts and autologous T lymphoblasts. Moreover, the CTL clones lacked NK activity as measured against K562 and Daudi cells. Panel studies indicated that the CTL clones also killed approximately 50% of the allogeneic melanoma cells preferentially, whereas the corresponding T lymphoblasts were not lysed. Monoclonal antibodies against class I (W6/32) and class II (279) MHC antigens failed to block the reactivity of the CTL clones against O-mel and allogeneic melanoma cells, indicating that a proportion of human melanoma cells share determinants that are different from HLA antigens and that are recognized by CTL clones. In contrast to the CTL clones isolated from MLTC, the clones obtained from MLC also lysed JY cells, which initially were used as stimulator cells. The reactivity of O-36 against JY could be inhibited with W6/32, demonstrating that this reactivity was directed against class I MHC antigens. These results suggest that the lysis of O-mel and JY cells by O-36 has to be attributed to two independent specificities of this CTL clone. The specificity of the other cross-reactive CTL clone (O-1) could not be determined. The notion that individual CTL clones can have two specificities was supported by the following observations. The cytotoxic reactivity of both O-1 (T4+) and O-36 (T8+) against JY was blocked by monoclonal antibodies directed against T3 and human LFA-1, and against T3, T8, and human LFA-1, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)
来自一名黑色素瘤患者的外周血淋巴细胞（PBL）的激活，无论是在二级混合淋巴细胞培养（MLC）中（其中来自JY细胞系的EB病毒转化的B细胞用作刺激细胞），还是通过在混合白细胞肿瘤细胞培养（MLTC）中与自体黑色素瘤细胞共培养，都导致产生针对自体黑色素瘤（O-mel）细胞的细胞毒性活性。从这些活化的大量培养物中分离出四个克隆的细胞毒性T淋巴细胞（CTL）系。CTL克隆O-1（T3 +、T4 +、T8 -、OKM-1 -、HNK -、HLA-DR +）和O-36（T3 +、T4 -、T8 +、OKM -、HNK -、HLA-DR +）是从MLC中获得的，而CTL克隆O-C7（T3 +、T4 +、T8 -、OKM-1 -、HNK -、HLA-DR +）和O-D5（T3 +、T4 -、T8 +、OKM-1 -、HNK、HLA-DR +）是从自体MLTC中分离出来的。所有四个CTL克隆对O-mel细胞都具有强烈的细胞毒性，但不能裂解自体成纤维细胞和自体T淋巴母细胞。此外，根据对K562和Daudi细胞的检测，CTL克隆缺乏NK活性。分组研究表明，CTL克隆还优先杀死了约50%的同种异体黑色素瘤细胞，而相应的T淋巴母细胞未被裂解。针对I类（W6/32）和II类（279）MHC抗原的单克隆抗体未能阻断CTL克隆对O-mel和同种异体黑色素瘤细胞的反应性，表明一部分人黑色素瘤细胞具有与HLA抗原不同的决定簇，并且能被CTL克隆识别。与从MLTC中分离的CTL克隆不同，从MLC中获得的克隆也能裂解最初用作刺激细胞的JY细胞。O-36对JY的反应性可以被W6/32抑制，表明这种反应性是针对I类MHC抗原的。这些结果表明，O-36对O-mel和JY细胞的裂解必须归因于该CTL克隆的两种独立特异性。另一个交叉反应性CTL克隆（O-1）的特异性无法确定。以下观察结果支持了单个CTL克隆可以具有两种特异性的观点。O-1（T4 +）和O-36（T8 +）对JY的细胞毒性反应分别被针对T3和人淋巴细胞功能相关抗原-1（LFA-1）以及针对T3、T8和人LFA-1的单克隆抗体阻断。
