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为探索进一步纯化肿瘤浸润淋巴细胞（ＴＩＬ）的方法及其体外抗瘤作用特点，为临床应用ＴＩＬ治疗肺癌提供依据，用不连续密度梯度离心法从接种于Ｃ５７ＢＬ／６小鼠体内的Ｌｅｗｉｓ肺癌（ＬＬＣ）瘤体中分离提取ＴＩＬ，过夜贴壁，剔除贴壁细胞后，可明显提高ＴＩＬ的纯度。纯化的ＴＩＬ在含１０００Ｕ／ｍｌＩＬ２的完全培养基中继续培养。免疫荧光表型分析发现，ＴＩＬ中ＣＤ＋４、ＣＤ＋８初为０９１±０１２，培养后为０８３±００７，体外ＴＩＬ对ＮＫ敏感的Ｌ９２９细胞有一定的杀伤作用，效靶比为２００∶１和１００∶１时的杀伤率高于５０∶１（Ｐ＜００５），表明ＬＬＣ浸润淋巴细胞体外的杀伤活性具有一定的非特异性。
In order to explore the method of further purification of tumor infiltrating lymphocytes (TIL) and its in vitro anti-tumor effects, it provides a basis for the clinical application of TIL in the treatment of lung cancer, and Lewis lung cancer inoculated into C57BL/6 mice by discontinuous density gradient centrifugation ( TIL was isolated and extracted from tumors, and adhered to the wall overnight. After removing adherent cells, the purity of TIL was significantly increased. Purified TIL was further cultured in complete medium containing 1000 U/ml IL-2. Immunofluorescence phenotype analysis revealed that CD+4 and CD+8 in TIL were initially 0.991±0.12, and after culture they were 083±007. In vitro, TIL had a certain killing effect on NK-sensitive L929 cells. The killing rate at 200:1 and 100:1 was higher than 50:1 (P < 0.05), indicating that the in vitro killing activity of LLC infiltrating lymphocytes has certain non-specificity.