【摘要】 目的探讨负载自体肿瘤细胞裂解物的成熟树突状细胞(ATLs-mDCs)体外诱导个体化抗胃癌过继免疫治疗的效应。方法建立短期培养的原代胃癌细胞系。用ATLs-mDCs激活自体T细胞,制备肿瘤特异性细胞毒性T细胞(CTLs)。自体树突状细胞(DCs)均分为未成熟DCs、成熟DCs和ATLs-mDCs 3组,分别应用流式细胞仪及混合淋巴细胞增殖反应方法,检测DCs的免疫功能状态;应用细胞毒杀伤试验验证肿瘤特异性CTLs的杀伤活性;应用酶联免疫吸附试验(ELISA)测定DCs培养上清中白细胞介素12(IL-12)和CTLs上清中γ干扰素(IFN-γ)的分泌水平。结果ATLs-mDCs上调HLA-DR、CD80、CD83和CD86的表达水平,同时获得高效刺激自体T细胞增殖的能力。ATLs-mDCs诱导产生的CTLs对自体胃癌细胞的杀伤率为(84±11)%,显著高于对两株异体胃癌细胞的杀伤率[(19±7)%和(19±11)%(t=54.18和56.46,P值均<0.01)]。ATLs-mDCs上清液中IL-12的浓度显著高于单纯成熟DCs(t=15.47,P<0.01)及未成熟DCs(t=28.44,P<0.01)。3组DCs分别激活自体T细胞产生的CTLs上清液中INF-γ的浓度ATLs-mDCs组高于单纯成熟DCs组(t=4.84,P<0.05),并显著高于未成熟DCs组(t=13.74,P<0.01)。结论ATLs-mDCs在体外诱导产生的CTLs能有效特异性杀伤自体胃癌细胞。更多还原

【Abstract】 Objective To investigate the antitumor effects of cytotoxic T lymphocytes(CTLs) induced by autologous dendritic cells that were inspired by autologous tumor lysates(ATLs-mDCs).Methods(Primary gastric) cancer cells prepared by short-term culture were used as targets.ATLs-mDCs were subjected to activate autologous T cells to generate CTLs.The immunological functions of DCs were evaluated by flow cytometry and by mixed leukocyte response(MLR) assay.The antitumor outcome of tumor antigen specific CTLs was tested by cytotoxicity assay.Concentrations of IL-12 in cultured DCs and INF-γ in CTLs were measured by ELISA.Results The expressions of MHC-Ⅱ,CD80,CD83 and CD86 were significantly up-regulated in ATLs-mDCs,moreover,the ATLs-mDCs obtained the capability of stimulating the proliferation of autologous T cells with high efficiency.The secretion of IL-12 in ATLs-mDCs was significantly higher than that in pure mature DCs(t=15.47,P<0.01) and in immature DCs(t=28.44,P<0.01).The secretion of INF-γ in CTLs activated by ATLs-mDCs was significantly higher than that in CTLs by pure mature DCs((t=4.84,)P<0.05) and in CTLs by immature DCs(t=13.74,P<0.01).The antigen specific cytotoxicity of CTLs induced by ATLs-mDCs was significantly higher against autologous tumor cells([(84±11)%]) than that against two allogeneic tumor cell lines [(19±7)% and(19±11)%;t=54.18 and 56.46,P<0.01,respectively].Conclusions ATLs-mDCs might mediate the antigen specific CTLs against autologous gastric cancer cells ex vivo with high efficiency.更多还原