对8例晚期恶性肿瘤的肿瘤浸润淋巴细胞(TIL)分离、培养至10~9细胞以上,再重输给病人,同时应用小剂量IL2。在治疗前一周及治疗后一个月采集病人外周静脉血、分离单个核细胞作为效应细胞,选用一组不同类型人肿瘤细胞株为靶细胞,以MTT比色法测定效应细胞对靶细胞的杀伤活性,同时分析效应细胞表型变化,结果显示,治疗后对不同靶细胞的杀伤活性均有不同程度提高。最有趣的是对与病人有相同组织学类型的瘤细胞的杀伤活性显著提高(43升高到1249溶解单位p<0.05),用T4、T8单抗双染流式细胞仪分析表明T4、T8细胞表面荧光强度明显增强(T4∶1.9→3.98,T8∶4.45→7.2 p<0.05)。提示TIL能够明显上调病人的免疫状况,其作用途径除直接的细胞毒效应外,免疫调节作用可能是其主要效应途径。

Tumor infiltrating lymphocytes(TILs) were isolated by enzymatic digestion and discontinuous gradient centrifugation from 8 human advanced tumors (4 stomach carcinoma, 2 liver cancer, 1 non-small-cell lung carcinoma and 1 colon cancer). These cells were cultured in complete RPMI 1640 medium supplemented with l000U/ml of rhIL2 for 4-6 weeks, till the cell number reach over l09/total, reinfused to the same patients i.v. meanwhile, the patients received 105U of rhIL2 i.m for 5 days. One week before and one month after TIL infusion periphery blood from the patients was collected and the mononuclear cells were isolated. Cytotoxicity against a panel of tumor cell targets by MTT colorometric assay and lymphocyte phenotype by two-color flow cytometry were mornitored. The results showed that there was significant increase in the killing ability to the tested tumor targets to different extent, especially the killing to the target cells which shared the same histological type with the patients tumor. (43 against 1249 lytic units p<0.05), an enhancement of fluoreceint intensity on CD4 and CD8 cell subsets. ( T4 1.92 to 3.98,4.45 to 7.2 p<0.05) All the findings indicate that TILs plus lower dose rhIL2 are able to up-regulate the cellular immunological state in patients through immunological regulatory effect by cytokine cascade addition to the direct cytotoxicity.