葡萄球菌肠毒素A对肝癌肿瘤浸润淋巴细胞抗瘤活性的作用

Antitumor efficacy of tumor infiltrating lymphocytes from hepatocellular carcinoma stimulated by staphylococcal enterotoxin A in vitro

目的探讨葡萄球菌肠毒素A(SEA)对肝癌肿瘤浸润淋巴细胞(TIL)抗瘤活性的诱导作用。方法取5例手术切除肝癌标本,分离TIL,在SEA作用下进行培养。定时记数,了解其增殖情况。流式细胞仪检测其CD3、CD4、CD8表达情况,噻唑蓝(MTT)比色法测定其对HepG-2肝癌细胞株的细胞毒活性,酶联免疫吸附试验(ELISA)测定培养上清液中肿瘤坏死因子(TNF)-α和γ-干扰素(IFN-γ)浓度。结果在SEA刺激下,2周TIL扩增100倍。1周后CD3+细胞占95%以上,CD8+细胞较CD4+细胞增殖更迅速。TIL细胞毒活性随培养逐渐增强。在培养的前10d内,TIL产生大量的TNF-α峰值达(453.70±9.26)ng/L和IFN-γ,其峰值达(2013.22±20.41)ng/L。结论SEA可高效、迅速诱导肝癌TIL的抗瘤活性。

Objective To evaluate in vitro the antitumor effects of tumor infiltrating lymphocytes (TIL) from the hepatocellular carcinoma tissues in response to the staphylococcal enterotoxin A (SEA).Methods TIL were successfully isolated from tumor tissues of 5 HCC patients.SEA was tested in vitro for its ability to stimulate TIL to acquire antitumor reactivity.The proliferation and cytotoxicity of TIL were observed.Flow cytometer was used to detect the expression of CD3,CD4 and CD8 on the surface of TIL.The TNF-α and IFN-γ secreted in the culture medium were measured by the method of ELISA.The results were compared with those in control group (TIL stimulated by IL-2).Results Being cultured with additive SEA,there was a sharp rise in number of T cells,especially CD8^+ cells.Cytotoxic activity was increased.TNF-α and IFN-γ were secreted predominantly in the first week.The peak levels of TNF-α and IFN-γ were (453.70± 9.26) ng/L and (2?013.22± 20.41) ng/L respectively.Conclusion SEA might be a potential stimulus to TIL.The antitumor efficacy of TIL induced by SEA was stronger than that by IL-2.