自体胸水上清培养肿瘤浸润性淋巴细胞生长所需白细胞介素-2的适宜剂量

目的：模拟体内环境,摸索出应用自体胸水上清培养肿瘤浸润性淋巴细胞（TIL）时所需白细胞介素-2（IL-2）的最适剂量。方法：恶性胸水中的TIL经贴壁法分离后,分成A、B、C三组,分别应用含有细胞因子IL-2、CD3单克隆抗体（OKT3）及外源凝集素（PHA）的自体胸水上清进行培养,其中A组IL-2终浓度为6000u/ml;B组IL-2首剂终浓度为6 000u/ml,其后换液时改为1000u/ml;C组IL-2终浓度为1000u/ml,比较不同剂量IL-2培养的TIL增殖速率、培养前后免疫表型变化和对自体肿瘤细胞的杀伤活性。结果：培养的第3、7、14和21 d检测发现,三组TIL均发生明显扩增,其中A、B两组间TIL的生长速度无差异,但均明显快于C组;TIL免疫表型变化和对自体肿瘤细胞杀伤活性经组间比较无显著差异。结论：TIL培养时,首剂IL-2的终浓度6000u/ml有着非常重要的意义,它有助于早期解除TIL的免疫抑制状态,使其在培养的第3 d就发生明显的活化和扩增,为自体TIL体外分离后仅作短期培养即回输胸腔,后续胸腔内注入IL-2,使其进一步生长,发挥杀瘤效应,从而为治疗恶性胸水奠定了理论基础。

Appropriate dosage of interleukin-2 for tumor infiltrating lymphocyte cultured in self-supernatant of malignant pleura fluid

Objective:To select an appropriate dosage of IL-2 for tumor infiltrating lymphocyte(TIL).Methods:Isolated by the attachment method we established,TIL was cultivated in self-supernatant of malignant pleura,with three different concentrations of IL-2,such as 6000u per ml,6000u per ml for the first administration followed by 1000u per ml,or 1000u per ml.The expansion,killing activity and phenotype changes of TIL cultured in different cultures were assessed.Results:As cultured in self-supernatant of malignant pleura fluid,the concentrations of IL-2,such as 6000u per ml or 6000u per ml for the first administration followed by 1000u per ml seemed benefit for TIL proliferation.Conclusion:6000u per ml of IL-2 for the first administration was very important.It could help TIL to activate and proliferate early.The study described here offers the possibility for TIL cultured in vitro self-supernatant of malignant pleura fluid in vitro for a short time,and then reinfuse to thorax for further expanding and controlling the malignant pleura effusion in the presence of IL-2,and thereby minimizing the risks of contamination.