脐血及外周血中细胞毒性T细胞的体外诱导及杀瘤活性研究

Study on Cytotoxity of CTL Cell From Umbilical Blood in Ovarian Cancer Cell Line

目的 在体外研究卵巢癌细胞诱导脐血来源的CTL的扩增及其杀瘤活性。探索其用于卵巢癌过继免疫治疗的可行性。方法 从脐血中分离出单个核细胞 ,用不同剂量的SKOV 3细胞及IL -2刺激扩增 ;用FCM检测表型变化 ,MTT比色法检测其对SKOV 3及K 5 62的杀伤活性 ,并与用同样方法所得外周血来源CTL细胞相对照。结果 用 2× 10 6个 /ml的卵巢癌细胞可诱导脐血CTL细胞较高的杀伤活性为 5 3 .46%± 8.45 % ;诱导 10天后杀伤SKOV 3活性为 44 .0 9%± 6.95 % ,杀伤K 5 62的活性为41.0 3 %± 5 .79% ,CD3、CD4、CD 8及CD4/CD8值分别为 5 2 .0 0± 9.5 0、3 6.0 0± 4.80、3 8.0 0± 10 .2 0、1.10± 0 .40。结论 应用适当剂量的抗原可诱导较高活性的脐血源CTL 。

Objective To study the proliferation and activity of CTL from umbilical blood which induced by inactivated ovarian cancer cell line SKOV3.Investigate to some valuable reference of CTL from cord blood as adoptive immunotherapy for ovarian cancer.Methods Mononuclear Cell separated from umbilical blood(UBMC) by Ficoll.UBMC was stimulated by inactivated SKOV3 and IL-2.Assay the phenotypes(CD3?CD4?CD8) by Flow cytometry.Assay the activity of kill K562 or SKOV3 by MTT colometric.The pheripheral blood Mononuclear cells as the contral.Results The activity of CTL cells from cod blood is the strongest(53.46±8.45)% when the induced ovarian tumor cell is 2×10 6/ml.Observation on cytotoxity against SKOV3 and K562 cells shows that CTL from cord blood have higher activity(44.09±6.95)% than NK(41.03±5.79)% on tenth day.On tenth day,the positive rates of CD3?CD4?CD8 and CD4/CD8 of CTL cells is 52.00±9.50?36.00±4.80?38.00±10.21 and 1.10±0.40,respectively.Conclusion These research have shown that higher activity of CTL from cord blood can be induced by suitable antigen doses.CTL cells from cord blood have obvious cytotoxity against ovarian tumor in vitro.