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本文从12例骨肉瘤患者手术切除的实体瘤中分离TIL,从患者外周血中分离淋巴细胞,以4小时~(51)cr 释放试验测定经rI1-2(500～1000U/m1)激活后的TIL 和LAK 细胞体外抗瘤活性及其特异性,并在培养的不同时间对TIL 表型进行分析。结果表明:12例骨肉瘤患者的TIL 培养10～20天,对K_(562)和LiBr靶细胞(效靶比25:1)平均杀伤活性分别为50.6±12.6%和42.6±10.2%。LAK 细胞对K_(562)和LiBr 细胞的杀伤活性分别为47.4±12.3%和37.6±8.5%。二者对K_(562)和LiBr 细胞的杀伤活性相差不显著(P>0.05)。其中7例患者的TIL 和LAK 细胞对自身肿瘤细胞杀伤活性分别为39.9±7.9%和26.3±6.8%,二者相差显著(P<0.01).TIL 表型特征为,CD3~+细胞比例在培养期间无明显变化,CD4~+细胞比例随培养时间延长有逐渐增加趋势.CD8~+细胞比例有逐渐减少趋势。
In this paper, TIL was isolated from 12 solid tumors resected from patients with osteosarcoma. Lymphocytes were isolated from peripheral blood of patients and measured after activation with rI1-2 (500-1000 U/m1) in a 4-hour (51) cr release test. The anti-tumor activity and specificity of TIL and LAK cells in vitro, and analysis of TIL phenotype at different time of culture. The results showed that in 12 cases of osteosarcoma TIL cultured for 10 to 20 days, the average killing activity of K_(562) and LiBr target cells (Effect ratio=25:1) was 50.6±12.6% and 42.6±10.2%, respectively. The killing activity of LAK cells against K562 and LiBr cells was 47.4±12.3% and 37.6±8.5%, respectively. The killing activity of K_(562) and LiBr cells was not significantly different (P>0.05). Among them, the killing activities of TIL and LAK cells on autologous tumor cells were 39.9±7.9% and 26.3±6.8%, respectively, which were significantly different (P<0.01). The TIL phenotype was characterized by the proportion of CD3~+ cells in culture. There was no significant change during the period, and the proportion of CD4~+ cells gradually increased with the extension of culture time. The proportion of CD8~+ cells gradually decreased.