Keyaki A, Kuribayashi K, Sakaguchi S, Masuda T, Yamashita J, Handa H, Nakayama E
Immunology. 1985 Sep;56(1):141-51.
We demonstrated the efficacy of a long-term cultured cytotoxic T-lymphocyte line, CTLL-D4, on tumour growth inhibition using athymic nude mice as recipients. CTLL-D4, specific for a unique surface determinant on a radiation-induced leukaemia RL male 1 of BALB/c origin, was obtained from the limiting dilution culture of MLTC cells performed between spleen cells of a CB6F1-nu/+ mouse immunized in vivo and RL male 1 stimulator cells, and cultured for several months in the absence of added TCGF as described in our preceding paper (Kuribayashi, 1985). The specific inhibition of tumour growth by CTLL-D4 was demonstrated both in Winn-type neutralization assay and in systemic transfer experiments. A subcutaneous inoculation of the mixture of CTLL-D4 and RL male 1 cells resulted in the complete inhibition of tumour growth, even at the effector to tumour cell ratio of 1:1, whereas non-cytolytic D4f, which was self-Ia antigen(s)-reactive, composed entirely of Lyt-1+23- T cells and derived originally from CTLL-D4 but completely lost its cytotoxic activity during culture with the irradiated syngeneic feeder cells alone, had no inhibitory effect at all. In the adoptive transfer studies, the subcutaneously established tumours were rejected by the single i.v. transfer of 2 X 10(7) CTLL-D4 cells into CB6F1-nu/nu mice. However, D4f was ineffective again in this systemic transfer system. When the effect of CTLL-D4 cells on tumour rejection in vivo was compared to that of non-cultured spleen cells hyperimmunized with RL male 1 cells, the former exhibited more rapid rejection in nude mice after i.v. transfer than the latter did, suggesting that CTLL-D4 cells also attack the tumour cells much more effectively as effectors in vivo. Thus, it is conceivable that CTLs are mainly involved in tumour rejection in this adoptive transfer system using RL male 1 tumour cells and athymic nude mice.
我们以无胸腺裸鼠为受体，证明了长期培养的细胞毒性T淋巴细胞系CTLL-D4对肿瘤生长的抑制作用。CTLL-D4对源自BALB/c的辐射诱导白血病RL雄性1的独特表面决定簇具有特异性，它是通过在体内免疫的CB6F1-nu/+小鼠的脾细胞与RL雄性1刺激细胞之间进行的MLTC细胞有限稀释培养获得的，并按照我们之前的论文（栗林，1985年）所述，在不添加TCGF的情况下培养了几个月。在Winn型中和试验和全身转移实验中均证明了CTLL-D4对肿瘤生长的特异性抑制作用。皮下接种CTLL-D4和RL雄性1细胞的混合物，即使效应细胞与肿瘤细胞的比例为1:1，也能完全抑制肿瘤生长，而不具有细胞溶解活性的D4f，它对自身Ia抗原具有反应性，完全由Lyt-1+23-T细胞组成，最初源自CTLL-D4，但在仅与经辐射的同基因饲养细胞培养期间完全丧失了细胞毒性活性，却完全没有抑制作用。在过继转移研究中，皮下建立的肿瘤通过向CB6F1-nu/nu小鼠单次静脉注射2×10(7)个CTLL-D4细胞而被排斥。然而，D4f在这个全身转移系统中再次无效。当将CTLL-D4细胞对体内肿瘤排斥的作用与用RL雄性1细胞超免疫的未培养脾细胞的作用进行比较时，前者在静脉注射后在裸鼠中表现出比后者更快的排斥反应，这表明CTLL-D4细胞作为效应细胞在体内对肿瘤细胞的攻击也更有效。因此，可以想象在这个使用RL雄性1肿瘤细胞和无胸腺裸鼠的过继转移系统中，细胞毒性T淋巴细胞主要参与肿瘤排斥反应。
