轮状病毒感染后先天性免疫RLRs通路部分关键因子Q-PCR检测方法建立

Establishment of Q-PCR detection method of RLRs signaling pathway after rotavirus infection

维甲酸诱导基因样受体(RLRs)能够识别细胞质中的病毒双链RNA,激活下游信号转导通路,最后促进I型干扰素分泌,对抗病毒感染。本试验针对牛轮状病毒UK株VP6、人IRF3及IRF7、RIG-I、MDA5基因序列设计特异性引物,从感染轮状病毒的Caco-2细胞中提取病毒总RNA,建立检测RLRs信号转导通路上述关键因子的Q-PCR方法。结果显示在感染早期各关键因子表达均有所上升,感染后期RIG-I及MDA5没有显著变化。本试验建立的检测方法可以检测宿主细胞感染轮状病毒后的先天性免疫应答反应,为是否产生免疫抑制提供数据支持。

Cytoplasmic pattern recognition receptors enable recognize the viral double stranded RNA, and followed by activating downstream signaling pathway, and finally promote the secretion of type I interferon to against viral infection. In this study, specific primers for bovine rota- virus UK strain VP6, human IRF3 and IRF7, RIG-I, MDA5 gene sequences were synthesized and a Q-PCR method to quantify some key factors of the RLRs signal transduction pathway in Caco-2 cells in- fected with UK rotavirus strains was established. The results demonstrated that the expression of all key factors increased during the early infection and significant change in RIG-I and MDA5 at during later stage of infection was not observed. The detection method established in this study can detect the innate immune response of host cells infected with rotavirus and provide data to elucidate whether immunosuppression is induced after viral infection to heterogeneous host cells.