【摘要】 目的观察卵巢癌(OC)患者肿瘤浸润淋巴细胞(TILs)的抗肿瘤活性。方法采用联合酶消化分离并提纯OC组织中的TILs和OC细胞,以IL-2体外诱导活化扩增TILs。活化前及活化第14天,采用流式细胞仪检测TILs免疫表型(CD+3、CD+4、CD+8、CD+4/CD+8);活化前及活化第3、7、14天,采用MTT法检测TILs对自体OC细胞、人白血病细胞株K562、人B细胞淋巴瘤细胞株Raji的杀伤活力;收集经活化第14天TILs作用前后的OC细胞培养上清液,ELISA法检测肿瘤分子标志物CA-125、CA-153、血管内皮生长因子(VEGF)、基质金属蛋白酶9(MMP-9)。结果与活化前比较,活化第14天TILs中CD+3、CD+4、CD+8细胞比例增加,CD+4/CD+8细胞比值降低;随着活化时间延长,TILs对三种靶细胞的杀伤率增高(P均<0.05),且对OC细胞的杀伤率高于其余2种细胞(P均<0.05);与处理前相比较,活化第14天TILs作用的OC细胞培养上清液中CA-125、VEGF、MMP-9水平均显著降低(P均<0.05)。结论卵巢癌患者的TILs具有抗肿瘤活性,低浓度IL-2可增强其特异性和非特异性抗瘤活性。更多还原

【Abstract】 Objective To observe the antitumor effect of tumor-infiltrating lymphocytes( TILs) in patients with ovarian cancer( OC). Methods The TILs and autologous OC cells were isolated and purified from OC tissues by using joint enzyme digestion method,then the TILs were activated and amplified in vitro by IL-2. The immunophenotypes( CD+3,CD+4,CD+8and CD+4/ CD+8) of TILs were tested by flow cytometry before the activation and the 14 th day of activation; the Methyl Thiazolyl Tetrazolium( MTT) was used to detect the cytotoxic effects of the TILs on autologous OC cells,human leukemia cell line K562 and human B lymphoma cell line Raji before activation and on the 3th,7th and 14 th day of activation; the culture supernatant of ovarian cancer was collected before and after the autologous OC cells were killed by TILs on14 th day,then,the tumor molecular markers CA-125,CA-153,vascular endothelial growth factor( VEGF) and matrix metalloproteinase 9( MMP-9) of culture supernatant were detected by using the enzyme-linked immunosorbent assay( ELISA). Results Compared with before activation,the proportions of CD+3,CD+4and CD+8of TILs were significant increased on the 14 th day of activation,while the CD+4/ CD+8ratio was significant decreased. With the prolonged activation time,the cytotoxic effects of TILs on the three kinds of target cells were significantly increased( all P < 0. 05),and the killing rate of OC cells was higher than the rest of two kinds of cells( all P < 0. 05). Compared with before treatment,the CA-125,VEGF and MMP-9 of autologous OC in the culture supernatant were significantly decreased on the 14 th day of activation( all P < 0. 05). Conclusion The TILs from OC tissues have antitumor activity,and the specificity and non-specificity cytotoxic function of TILs can be effectively activated and amplified by low concentration of IL-2.更多还原