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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 investigate the effect of staphylococcal enterotoxin A (SEA) on the antitumor activity of tumor infiltrating lymphocytes (TIL) in hepatocellular carcinoma. Methods Five cases of HCC were resected, and TIL was isolated and cultured under the action of SEA. Time counting, to understand its proliferation. The expression of CD3, CD4 and CD8 were detected by flow cytometry. The cytotoxicity of CD3, CD4 and CD8 on HepG-2 hepatoma cell line was determined by MTT colorimetric assay. Tumor necrosis factor (TNF) -alpha and interferon-gamma (IFN-γ) concentrations. Results The TIL was amplified 100-fold at 2 weeks after SEA stimulation. After 1 week CD3 + cells accounted for more than 95%, CD8 + cells proliferate more rapidly than CD4 + cells. TIL cytotoxic activity gradually increased with culture. During the first 10 days of culture, TIL produced a large number of TNF-α peaks (453.70 ± 9.26) ng / L and IFN-γ with peak values (2013.22 ± 20.41) ng / L. Conclusion SEA can efficiently and rapidly induce the anti-tumor activity of TIL of hepatocellular carcinoma.