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目的:观察研究人外周血单核细胞来源的树突状细胞(DC)转染含癌胚抗原(CEA)片段的重组腺相关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片断的重组腺相关病毒转染未成熟DC,诱导特异性T细胞。检测体外培养的DC和CTL活性,并使用MTT法检测细胞毒性T细胞(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DC高表达CD40、CD86、IL-12,诱导的细胞毒性T细胞高表达IFN-γ;转染后DC诱导特异性细胞毒性T细胞可有效识别并杀伤HLA-A11阳性的LOVO细胞。结论:重组腺相关病毒转染DC,不明显改变DC表型和刺激淋巴细胞增殖、分化功能,可诱导自体细胞毒性T细胞增殖,含CEA片断的腺相关病毒转染DC诱导自体细胞毒性T细胞对LOVO细胞有明显杀伤作用,DC疫苗可以作为直肠癌患者免疫治疗的有效补充。
AIM: To investigate the effect of specific T cells induced by dendritic cells derived from peripheral blood mononuclear cells (DCs) transfected with recombinant adeno-associated virus containing CEA fragments on rectal cancer cell lines LOVO and SW480 In vitro killing effect. Methods: Peripheral blood from healthy volunteers with HLA phenotype A11 was collected, monocytes were isolated and cultured in vitro. Immature DCs were transfected with recombinant adeno-associated virus containing CEA fragment to induce specific T cells. The activity of DC and CTL in vitro was detected. The killing effect of cytotoxic T lymphocytes (CTL) on LOVO cells was detected by MTT assay. Results: CD40, CD86 and IL-12 were highly expressed in transfected or untransfected mature DCs. IFN-γ was highly expressed in cytotoxic T cells induced by DCs. DC induced specific cytotoxic T cells after transfection could be effectively identified And killed HLA-A11-positive LOVO cells. CONCLUSION: DCs transfected with recombinant adeno-associated virus can induce autologous cytotoxic T cell proliferation without significantly changing the DC phenotype and stimulating the proliferation and differentiation of lymphocytes. DCs containing CEA fragments can induce autologous cytotoxic T cells LOVO cells have obvious killing effect, DC vaccine can be used as an effective supplement for immunotherapy of patients with rectal cancer.