含CEA片段的重组腺病毒转染DCs诱导特异性T细胞对结直肠癌细胞株的体外杀伤作用

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。

Specific anticolorectal carcinoma T cells generated by dendritic cells transferred with CEA gene mediated by adeno virus and its Inhibiting effect on colortal carcinoma cells

Objective:To observe the changes in the functions of human peripheral blood monocyte-derived dendritic cells(DCs) after CEA gene infection mediated by recombinant adeno virus type2(rADV) and the specific killing activity of the CTL induced by the specific DC vaccination to the LoVo and SW480 cells.Methods:Immature DCs were generated from peripheral blood monocyte cells of healthy volunteers with HLA-A11 positive and infected with the recombinant adeno virus carrying CEA gene or not.The surface markers such as CD40, CD 83, CD86 were detected by flow cytometry.The IL-12 released in the supernatnants of DCs and the IFN-γ released in the CTLs were assaged by ELISA detection kit. The specific killing activity of CTL to LOVO cells was checked by MTT.Results:In both groups the DCs showed highly expressed CD40, CD86 and IL-12 after the infection. DCs infected with rADV can induce specific CTL and the CTL can specifically recognize LOVO cells and kill them. Conclusion:DCs infected by the rADV are the same as naive DCs in eliciting the differentiation of the lymphocytes toward T helper type I cells, and the functions and the surface markers of DCs remain unaffected after the infection.