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目的:探讨葡萄球菌肠毒素A(SEA)对卵巢癌肿瘤浸润淋巴细胞(TIL)及其外周血淋巴细胞(PBL)抗瘤活性的诱导作用。方法:取10例卵巢癌伴腹水患者实体瘤、腹水及外周血标本,分离TIL和PBL。在SEA及IL-2作用下培养,定时计数,了解其增殖情况;流式细胞仪检测其CD3、CD4、CD8表达;噻唑蓝(MTT)比色法测定其对K562及自体肿瘤细胞的细胞毒活性;酶联免疫吸附试验(ELISA)测定培养上清液中TNF-a和IFN-γ浓度。结果:10例中8例成功分离实体瘤TIL、腹水TIL及PBL。(1)SEA刺激的实体瘤TIL、腹水TIL及PBL增殖速率明显较IL-2诱导组快(P<0.05),但增殖高峰后出现下降趋势,IL-2组未出现此现象;(2)CD3+CD4+及CD3+CD8+T表达率均明显上升,其中SEA诱导组比IL-2组增加比例明显(P<0.05),以SEA作用的CD3+CD8+T比例增加最快;(3)TIL对自体肿瘤细胞的杀伤活性明显高于对K562细胞的杀伤活性(P<0.05),PBL对自体肿瘤细胞的杀伤活性则明显低于对K562细胞的杀伤活性(P<0.05),SEA激活组比IL-2组杀伤率高(P<0.05);(4)各效应细胞分泌的TNF-a、IFN-γ分别在培养的第2天和第4天达到高峰,高峰后迅速下降,SEA诱导组在前10天明显高于IL-2诱导组(P<0.05)。结论:SEA可高效、迅速诱导卵巢癌TIL的抗瘤活性。
Objective: To investigate the effect of staphylococcal enterotoxin A (SEA) on the tumorigenicity of tumor-infiltrating lymphocytes (TILs) and peripheral blood lymphocytes (PBLs) in ovarian cancer. Methods: Ten cases of solid tumor, ascites and peripheral blood from patients with ovarian cancer and ascites were isolated and TIL and PBL were isolated. The cells were cultured under the action of SEA and IL-2, counted regularly and their proliferation was observed. The expressions of CD3, CD4 and CD8 were detected by flow cytometry. The cytotoxicity of K562 and autologous tumor cells was assayed by MTT assay Activity and the concentration of TNF-a and IFN-γ in culture supernatants were determined by enzyme linked immunosorbent assay (ELISA). Results: TIL, TIL and PBL in 8 cases of 10 cases were successfully separated. (1) The proliferation rate of TIL, TIL and PBL of solid tumor stimulated by SEA was significantly faster than that of IL-2 induction group (P <0.05), but decreased after the peak of proliferation, but not in IL-2 group. (2) (P <0.05), and the proportion of CD3 + CD8 + T increased obviously with SEA; (3) The expression of CD3 + CD8 + The killing activity of TIL to autologous tumor cells was significantly higher than that of K562 cells (P <0.05). The killing activity of PBL to autologous tumor cells was significantly lower than that of K562 cells (P <0.05) (4) The levels of TNF-a and IFN-γ secreted by various effector cells peaked on the 2nd and 4th day of culture respectively, then decreased rapidly after the peak, and SEA was induced Group in the first 10 days was significantly higher than the IL-2 induction group (P <0.05). Conclusion: SEA can efficiently and rapidly induce the anti-tumor activity of ovarian cancer TIL.