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目的:探讨人外周血单核细胞来源的树突状细胞转染含CEA片段的重组腺病毒后所诱导的特异性T细胞对直肠癌细胞株LOVO和SW480的体外杀伤作用。方法:抽取HLA表型为A11的健康志愿者外周血,分离单核细胞,体外培养,使用含CEA片断的重组腺病毒转染未成熟DCs,诱导特异性T细胞。检测体外培养的DCs和CTL活性,并使用MTT法检测CTL对LOVO细胞的杀伤作用。结果:转染或未转染的体外培养的成熟DCs高表达CD40、CD86,IL-12,诱导的CTL细胞高表达IFN-γ;转染后DCs诱导特异性CTL可有效识别并杀伤HLA-A11阳性的LOVO细胞。结论:重组腺病毒转染DCs,不明显改变DCs表型和刺激淋巴细胞增殖、分化功能,可诱导自体CTL增殖,含CEA片断的腺病毒转染DCs诱导自体CTL对LOVO细胞有明显杀伤作用。
Objective: To investigate the in vitro cytotoxicity of specific T cells induced by dendritic cells derived from human peripheral blood mononuclear cells transfected with recombinant adenovirus containing CEA fragment on LOVO and SW480 cells. Methods: Peripheral blood from healthy volunteers with HLA phenotype A11 was isolated, monocytes were isolated and cultured in vitro. Immature DCs were transfected with recombinant adenovirus containing CEA fragment to induce specific T cells. The DCs and CTL activities in vitro were detected. MTT assay was used to detect the killing effect of CTL on LOVO cells. Results: CD40, CD86 and IL-12 were highly expressed in cultured DCs transfected or untransfected, and IFN-γ was highly expressed in CTL cells transfected or untransfected. CTL induced by DCs could effectively recognize and kill HLA-A11 Positive LOVO cells. CONCLUSION: DCs transfected with recombinant adenovirus can induce autologous CTL proliferation without significantly changing the phenotype of DCs and stimulating the proliferation and differentiation of lymphocytes. The transfection of DCs with CEA fragments induces cytotoxicity of autologous CTL on LOVO cells.