C型凝集素CLEC4M对HCVcc易感性的影响

Influence of C-type lectin domain family 4,member M (CLEC4M) on the susceptibility of Huh7.5 cells to HCVcc

目的利用HCV体外培养细胞模型,观察C型凝集素CLEC4M对于HCVcc易感性的影响。方法构建GV166-CLEC4M慢病毒表达载体,脂质体转染293T细胞,获取的慢病毒转染Huh7.5细胞,获得稳定过表达CLEC4M的Huh7.5细胞系(即Huh7.5-CLEC4M),Western印迹和RT-PCR检测CLEC4M的过表达情况。利用HCVcc病毒液感染过表达CLEC4M的Huh7.5细胞系,并设置单克隆抗体干扰组及甘露聚糖干扰组,检测CLEC4M对HCVcc感染性的变化。结果 Huh7.5细胞系获得稳定过表达的CLEC4M。Huh7.5-CLEC4M获得对HCVcc的高度易感性。经鼠抗人CLEC4M单克隆抗体和甘露聚糖干扰后,Huh7.5细胞的感染效率较前降低。结论 C型凝集素CLEC4M影响细胞对HCVcc的易感性,可能与HCV的嗜肝性有关。

Objective To investigate the influence of the C-type lectin CLEC4M on susceptibility to HCV in cell culture （HCVcc）.Methods A GV166-CLEC4M lentiviral expression vector was constructed and transfected into 293T cells with lipofectamine.The Huh7.5 cells was transfected with the lentiviral to establish Huh7.5 cell lines with a stable CLEC4M overexpression （Huh7.5-CLEC4M）,which would be verified by reverse transcription-polymerase chain reaction and western blot.Then established Huh7.5-CLEC4M cell lines were infected by HCVcc.Meanwhile,two interference groups,monoclonal antibody interference group （treated with monoclonal antibody）and mannan interference group （treated with manna）were set up.The changes of CLEC4M expressions in two interference groups were detected. Results The Huh7.5-CLEC4M cell line was successfully established with high susceptibility to HCVcc.Compared to these in two interference group before treatment,expressions of CLEC4M in the Huh7.5-CLEC4M decreased after interference,so were infection rates of HCVcc.Conclusion C-type lectin CLEC4M could affect susceptibility of cells to HCV,which might be associated with HCV-related hepatic tropism.