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目的体外高效扩增肿瘤浸润淋巴细胞（ＴＩＬ），为进一步开展基因治疗肿瘤奠定基础。②方法采用机械分离、酶消化和不连续密度梯度离心法分离提取ＴＩＬ前体细胞；以ＩＬ－２作为诱导剂，采用液相三步扩增方法扩增培养，并分别用ＭＴＴ和ＩＦＡ法与对照的自体ＬＡＫ细胞比较细胞动力学。同时对Ｋ５６２细胞和自体肿瘤细胞（ＡＴＣ）进行杀伤试验以及表型检测。③结果从２８例实体瘤组织和肿瘤引流淋巴结中获取的ＴＩＬ前体细胞，２６例培养成功，其中２３例ＴＩＬ数量达到治疗数量级；ＴＩＬ细胞对ＡＴＣ杀伤作用强于自体ＬＡＫ细胞（ｔ＝１１．４，１８．９，Ｐ＜０．０１）。ＴＩＬ细胞表型特点为ＣＤ３，ＣＤ４，ＣＤ８淋巴细胞亚群。④结论ＴＩＬ细胞具有高效杀肿瘤细胞作用，ＴＩＬ细胞可作为基因治疗的理想载体细胞。
Objective To efficiently infiltrate tumor infiltrating lymphocytes (TIL) in vitro and lay a foundation for further gene therapy for tumors. 2 Methods TIL precursor cells were isolated and isolated by mechanical separation, enzyme digestion and discontinuous density gradient centrifugation; IL-2 was used as an inducer to amplify the cells using the three-step liquid phase amplification method, and the MTT and IFA methods were used respectively. Control autologous LAK cells compare cell kinetics. At the same time, K562 cells and autologous tumor cells (ATC) were killed and phenotyped. 3 Results TIL precursor cells obtained from 28 solid tumor tissues and tumor-draining lymph nodes were successfully cultured in 26 cases, of which 23 cases had a TIL number of therapeutic magnitude; TIL cells had a stronger ATC killing effect than autologous LAK cells (t=11. 4, 18.9, P < 0.01). The phenotype of TIL cells is CD3, CD4, and CD8 lymphocyte subsets. 4 Conclusions TIL cells have an effect of killing tumor cells efficiently, and TIL cells can serve as ideal carrier cells for gene therapy.