葡萄球菌肠毒素A脂质体激活肿瘤浸润淋巴细胞抗肿瘤活性的实验研究

Experimental Study on the Activity of Staphyloccocal Enterotoxin A Liposome for Inducing Cytotoxicity of TIL from Human Hepatocellular Carcinoma Against Tumor Cells

目的 研究葡萄球菌肠毒素 A(SEA)脂质体 (L- SEA)体外激活肿瘤浸润淋巴细胞 (TIL)抗肿瘤的活性。方法 从 5例原发性肝癌患者癌组织中分离培养 TIL,分别以 L- SEA、SEA和 IL- 2刺激后 ,观察 TIL 的增殖曲线 ,EL ISA法检测细胞因子的分泌 ,流式细胞仪检测细胞亚群的变化 ,MTT法检测 TIL 的肿瘤杀伤活性。结果 L- SEA、SEA及 IL- 2均能有效刺激 TIL 的增殖 ,最高增殖倍数分别为 4 2 .5、5 1、12 .5倍。除第 4 d L- SEA组 TIL 的IFN- γ水平低于游离 SEA组外 ,此两组的细胞因子分泌没有显著性差异 (P>0 .0 5 ) ,且都高于 IL- 2组。 L- SEA刺激后 ,TIL 的 CD8+细胞亚群增殖更快 ,CD4 +细胞 / CD8+细胞出现倒置。经过 L- SEA培养后的 TIL 能有效杀伤Hep G- 2肿瘤细胞。结论 L- SEA仍然具有游离 SEA的刺激 TIL 增殖、分泌细胞因子、杀伤肿瘤细胞的活性。

Objective To investigate the activity of Staphyloccocal enterotoxin A liposome (L-SEA) for inducing cytotoxicity of tumor infiltrating lymphocytes (TIL) against tumor cells. Methods TIL were isolated from the tumor tissues of five hepatocellular carcinoma patients. L-SEA, SEA and IL-2 were tested in vitro for their activity levels in stimulating TIL proliferation. The TNF-α and IFN-γ secretion and cytotoxicity of TIL against HepG-2 liver cancer cells were estimated by ELISA and MTT, respectively. Results Both L-SEA and SEA significantly stimulated the proliferation of TIL. The cytokine secretion of L-SEA group was significantly higher than that of IL-2 group (P<0.05). There was no significantly statistical difference in cytokine secretion between L-SEA group and SEA group (P>0.05) except that IFN-γ secretion of L-SEA group was lower than that of SEA group at day 4 (P<0.05). Both L-SEA and SEA had potent ability to induce TIL cytotoxicity against HepG-2 cells. And no significant difference was observed between these two groups(P>0.05). Conclusion These results suggest that L-SEA is as efficient as SEA in activating TIL.