Induction of cytotoxicity of human tumor-infiltrating lymphocytes against autologous tumor cells and its application for cancer therapy

1)采用白细胞介素-2（IL-2）培养的自体外周血淋巴细胞（PBL）诱导淋巴因子活化杀伤细胞（LAK）进行过继免疫治疗。通过动脉或腔内输注LAK细胞的局部给药比静脉输注的全身给药具有更好的治疗效果。2)人实体癌浸润细胞中有一半是T淋巴细胞，且CD8^+T淋巴细胞的比例大于PBL。肿瘤浸润淋巴细胞（TIL）中含有γ - T细胞，结直肠癌中大多数细胞为CD8^+ γ - T细胞，其谱系与PBL不同。肝细胞癌比其他人类实体癌含有更多的γ - δ TIL。3)单纯用IL-2培养TIL不足以使其在体外扩增，我们发现在LAK细胞诱导的条件培养基中加入20%的条件培养基可以有效扩增TIL。4)新鲜分离的TIL未检测到对肿瘤细胞的细胞毒性。TIL的细胞毒性在培养过程中逐渐增强，在2 ~ 3周时达到最大，在4 ~ 7周时消失。发现植物血凝素（Phytohemagglutinin， PHA）能够部分恢复培养TIL的细胞毒性。5)通过术前瘤内注射生物反应调节剂OK-432在体内激活晚期胃癌TIL，并将治愈性手术后患者的生存与对照组患者进行比较。虽然轻度TIL患者的生存率没有差异，但TIL的激活提高了中度或重度TIL患者的3年生存率。

1) Cancer patients were treated with the adoptive immunotherapy with lymphokine-activated killer (LAK) cells which were induced by culture of autologous peripheral blood lymphocytes (PBL) with interleukin-2(IL-2). Local administration by intraarterial or intracavital infusions of LAK cells produced a superior therapeutic efficacy to systemic administration by intravenous infusions.2)A half of tumor-infiltrating cells into human solid cancers consisted of T lymphocytes and the proportion of CD8^+T lymphocytes were greater than that of PBL.Tumor-infiltrating lymphocytes (TIL) contained gammadelta T cells and the majority of those cells in colorectal cancers were CD8^+ gammadelta T cells whose lineage was different from that of PBL.Hepatocellular carcinoma contained a greater number of gammadelta TIL than other human solid cancers.3)Culture of TIL with IL-2 alone was inadequate for their in vitro expansion, and we found that TIL effectively expanded in culture by addition of 20% conditioned medium derived from LAK cell induction.4)Cytotoxicity against tumor cells was notdetected in freshly isolated TIL.Cytotoxicity of TIL gradually increased in culture, reached its maximum at 2 to 3 weeks, and disappeared at 4 to 7 weeks. Phytohemagglutinin (PHA) was found to be capable of partial restoration of the diminished cytotoxicity of cultured TIL.5) TIL of advanced gastric cancer were activated in vivo by preoperative intratumoral injection with a biological response modifier, OK-432, and survival of patients after curative sugery was compared with control patients. Although there was no difference in survival of patients with scanty TIL,the activation of TIL improved 3-year survival in patients with TIL of a medium or over degree.