转染自体胃癌细胞总RNA的树突状细胞诱导个体化免疫治疗的体外实验

目的探讨转染自体胃癌细胞总RNA的树突状细胞(DC)体外介导抗胃癌的免疫效应。方法制备短期培养的原代胃癌细胞。用rhGM-CSF、rhIL-4和TNF-α体外诱导胃癌患者外周血单个核细胞(PBMC)中DC的发育和成熟,并转染自体肿瘤细胞总RNA,激活自体T细胞产生CTL,用CCK-8试剂盒检测CTL的杀伤活性。应用流式细胞术及混合淋巴细胞培养技术检测DC的免疫功能状态。用ELISA法测定IL-12和INF-γ的水平。结果转染自体肿瘤细胞总RNA的成熟DC,不仅可高表达MHC-I、II类分子及CD80、CD83和CD86协同刺激分子,并可获得高效刺激自体或异体T细胞增殖的能力。转染RNA的成熟DC,分泌IL-12的水平及其刺激产生的CTL培养上清液中INF-γ的水平显著高于单纯成熟DC及未成熟DC;且CTL对自体胃癌细胞的杀伤率显著高于异体组。结论转染自体胃癌细胞总RNA的成熟DC能够体外诱导产生对自体肿瘤细胞具有高度抗原特异性杀伤活性的CTL。

In vitro experimental study on individualised immunotherapy induced by dendritic cells transfected with total RNA of autologous gastric cancer cells

AIM: To explore the efficiency of antitumor immunity induced by autologous dendritic cells (DCs) transfected with total RNA of autologous gastric cancer cells. METHODS: Short-term cultured primary gastric cancer cells were prepared. DCs in peripheral blood mononuclear cells (PBMCs) from gastric cancer patients were induced with rhGM-CSF, rhIL-4 and TNF-alpha. The mature DCs transfected with total RNA of autologous gastric cancer cells were subjected to activate autologous T cells transforming into CTLs, and the activity of CTLs was detected by using CCK-8 kit. The immunological function of DCs were evaluated by flow cytometry and mixed lymphocyte culture(MLC) assay. The levels of IFN-gamma and IL-12 were detected by ELISA. RESULTS: Mature DCs transfected with total RNA of autologous gastric cancer cells not only highly expressed costimulatory molecules (CD80, CD83 and CD86) and (MHC-I and MHC-II), but also powerfully stimulated allogenic or autologous T cell proliferation. The level of IL-12 secreted by mature DCs transfered with tumor RNA was notably higher than those secreted by untransfered and immature DCs, and the rate of killing autologous gastric cancer cells by CTLs was markedly higher than that of killing allogenic tumor cells. CONCLUSION: Mature DCs transfected with autologous gastric cancer cell total RNA can induce and activate high antigen-specific CTLs directed at autologous gastric cancer cells in vitro.