Takagi S, Chen K, Schwarz R, Iwatsuki S, Herberman R B, Whiteside T L
Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania.
Cancer. 1989 Jan 1;63(1):102-11. doi: 10.1002/1097-0142(19890101)63:1<102::aid-cncr2820630117>3.0.co;2-t.
Tumor-infiltrating lymphocytes (TIL) were isolated from 40 of 51 consecutive human liver tumor samples (primary hepatocellular carcinoma, 16 of 18; metastatic, 23 of 29; benign, one of four). Functional and phenotypic characteristics of fresh and recombinant interleukin-2 (rIL-2)-expanded TIL were evaluated. The expansion of TIL from hepatic tumors in the presence of 1000 units/ml of rIL-2 was possible in 60% of cases. In comparison to TIL from metastatic liver tumors, TIL obtained from primary liver tumors expanded faster and better in rIL-2 cultures. Expanded TIL from primary tumors had significantly higher cytotoxicity against K562 targets, but not Raji targets, than those from metastatic tumors. Cytotoxicity against fresh autologous tumor targets was detected in seven of eight cultures tested. TIL from primary tumors retained antitumor reactivity significantly longer in culture. The optimal in vitro cytotoxicity was achieved between days 20 and 60 of culture in the presence of rIL-2. Antitumor activity was associated with the increase in these TIL cultures of a cell population expressing the Leu19 antigen with or without the CD3 antigen. The frequency of the CD3+Leu19+ population showed a bimodal distribution during culture: the first peak of CD3+Leu19+ cells occurred between days 30 and 60 and was associated with the increased antitumor activity; the second peak occurred after day 60 and was not associated with activity. These findings demonstrate that TIL from most human hepatic tumors can be successfully isolated, cultured in rIL-2, and enriched in Leu19+ effectors. In addition, these TIL upon IL-2 activation in vitro are capable of lysing fresh autologous and/or allogeneic tumor targets.
从51例连续的人类肝脏肿瘤样本（原发性肝细胞癌18例中的16例；转移性肝癌29例中的23例；良性肿瘤4例中的1例）中的40例分离出肿瘤浸润淋巴细胞（TIL）。对新鲜的和重组白细胞介素-2（rIL-2）扩增的TIL的功能和表型特征进行了评估。在1000单位/毫升rIL-2存在的情况下，60%的病例中肝脏肿瘤的TIL能够扩增。与转移性肝脏肿瘤的TIL相比，原发性肝脏肿瘤的TIL在rIL-2培养中扩增得更快且更好。原发性肿瘤扩增的TIL对K562靶细胞的细胞毒性显著高于转移性肿瘤的TIL，但对Raji靶细胞无此差异。在测试的8个培养物中有7个检测到对新鲜自体肿瘤靶细胞的细胞毒性。原发性肿瘤的TIL在培养中保留抗肿瘤反应性的时间明显更长。在rIL-2存在的情况下，培养20至60天之间可达到最佳体外细胞毒性。抗肿瘤活性与这些TIL培养物中表达Leu19抗原（有或无CD3抗原）的细胞群体增加有关。CD3+Leu19+群体的频率在培养过程中呈双峰分布：CD3+Leu19+细胞的第一个峰值出现在第30至60天之间，与抗肿瘤活性增加相关；第二个峰值出现在第60天之后，与活性无关。这些发现表明，大多数人类肝脏肿瘤的TIL能够成功分离，在rIL-2中培养，并富集Leu19+效应细胞。此外，这些TIL在体外经IL-2激活后能够裂解新鲜自体和/或同种异体肿瘤靶细胞。
