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目的 研究腺相关病毒(AAV)为载体的含有乙型肝炎病毒(HBV)C基因重组病毒(rAAV-HBV-C)感染树突状细胞(DC)的效率及对DC生长和成熟的影响。 方法 从正常人外周血中分离单个核细胞收取单核细胞进行体外培养,分别用rAAV-HBV-C和293细胞裂解物感染刺激单核细胞,感染后的单核细胞在粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素4(IL-4)和肿瘤坏死因子α(TNF α)存在的条件下继续培养获得成熟的DC。用逆转录聚合酶链反应及流式细胞仪细胞内染色检测目的基因(HBV-C)的转录及表达;通过观察不同时间DC的形态及检测收获DC时其表面分化抗原(CD)的表达来评价DC的生长和成熟状态。 结果 rAAV-HBV-c感染后的DC可转录HBV-C基因并表达HBV-C抗原,病毒感染组与对照组收获的DC在细胞形态与CD表达方面差异无统计学意义。 结论 rAAV-HBV-C可有效感染DC,感染的病毒对DC的生长及成熟没有显著影响。
Objective To investigate the effect of adeno-associated virus (AAV) -containing dendritic cells (DCs) infected with hepatitis C virus (C) recombinant C virus (rAAV-HBV-C) on DC growth and maturation. Methods Mononuclear cells were isolated from normal human peripheral blood and mononuclear cells were collected for in vitro culture. Monocytes were infected with rAAV-HBV-C and 293 lysate, respectively. The mononuclear cells were infected in granulocyte-macrophages Mature DCs were cultured in the presence of GM-CSF, IL-4 and TNFα. The transcription and expression of target gene (HBV-C) were detected by reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry (FCM). The morphological changes of DCs were observed at different time points and the expression of surface differentiation antigen (CD) Evaluation of DC growth and maturation status. Results DCs transfected with rAAV-HBV-c could express HBV-C gene and express HBV-C antigen. There was no significant difference in cell morphology and CD expression between DCs infected with rAAV-HBV-c and controls. Conclusions rAAV-HBV-C can effectively infect DC, and the infected virus has no significant effect on the growth and maturation of DC.