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目的研究转染存活素基因对树突状细胞(DC)免疫功能的影响,观察修饰后的DC在体外诱导的抗消化道肿瘤免疫效应。方法脂质体介导存活素基因转染入DC,用蛋白印迹法检测培养上清存活素的表达,检测转存活素基因DC分泌细胞因子白细胞介素12(IL12)、肿瘤坏死因子α(TNFα)的功能,以及经流式细胞仪检测DC表面CD1a、CD83、MHCⅡ、CD80、CD86表达的高低,用噻唑蓝(MTT)法诱导人特异的细胞毒性T淋巴细胞(CTL)的能力。结果培养上清中均可以检测到存活素表达;转存活素基因DC的上清IL12、TNFα含量分别为(2652±327)pg/ml和(4371±835)pg/ml,比单纯DC组高(P<005);CD1a、CD83、MHCⅡ、CD80、CD86等在单纯DC表面低表达,在转基因DC表面高表达;MTT法检测,经转染存活素基因的DC提呈的细胞对胃癌细胞、结肠癌细胞、胆管癌细胞杀伤率分别为65%、77%、85%,而单纯DC杀伤作用较低。结论存活素基因转染修饰的DC能诱导细胞毒性T淋巴细胞的特异性,显著地提高DC的抗原提呈功能,体外能诱导高效而特异的抗癌免疫效应。
Objective To investigate the effect of transfection survivin gene on immune function of dendritic cells (DCs) and to observe the anti-digestive tract immune response induced by modified DCs in vitro. Methods The liposome - mediated Survivin gene was transfected into DC. The expression of Survivin in supernatant of culture supernatant was detected by Western blotting. The levels of interleukin - 12 (IL12), tumor necrosis factor - α (TNFα) ), And the ability of inducing human-specific cytotoxic T lymphocytes (CTL) by flow cytometry (FCM) was used to detect the expression of CD1a, CD83, MHCⅡ, CD80 and CD86 on DCs. Results Survivin expression was detected in the culture supernatant. The contents of IL12 and TNFα in supernatant of survivin gene DC were (2652 ± 327) pg / ml and (4371 ± 835) pg / ml, respectively, (P0.005). The expression of CD1a, CD83, MHCⅡ, CD80 and CD86 on the surface of pure DCs was low and was highly expressed on the surface of transgenic DCs. Colon cancer cells, cholangiocarcinoma cell killing rates were 65%, 77%, 85%, while the role of DC alone was low. Conclusion Survivin gene transfected DCs can induce cytotoxic T lymphocyte specificity, significantly improve DC antigen presenting function, in vitro can induce efficient and specific anti-cancer immune effect.