组织蛋白酶S抑制塞内卡病毒在IBRS-2细胞中的复制

The replication of Seneca Valley virus in IBRS-2 cells was inhibited by cathepsin S

【目的】本研究旨在探究猪源组织蛋白酶S(cathepsinS,CTSS)对塞内卡病毒(SenecaValley virus,SVV)复制的影响。【方法】SVV感染IBRS-2细胞,采用RT-qPCR在转录水平探究SVV感染对内源性CTSS表达的调控;采用ELISA测定SVV感染对CTSS酶活性的影响;通过Western blotting(WB)和RT-qPCR检测过表达CTSS对SVV复制及SVV诱导的抗病毒细胞因子的调控作用;合成针对CTSS的特异性siRNA,利用WB和RT-qPCR检测siRNA对CTSS的干扰效果以及CTSS被干扰后对SVV复制的影响。【结果】结果表明SVV感染IBRS-2细胞能显著上调内源性CTSS表达并增强CTSS酶活性;过表达CTSS能显著抑制SVV在IBRS-2细胞中的复制,同时促进宿主抗病毒细胞因子的表达;siRNA-2947下调内源性CTSS表达进而促进SVV复制。【结论】CTSS通过增强宿主抗病毒细胞因子上调表达而抑制SVV复制,本研究为进一步深入探究宿主CTSS在抗SVV免疫应答中的作用及机制提供参考依据。

[Objective]The purpose was to explore the effect of porcine cathepsin S(cathepsin S,CTSS)on SVV(Seneca Valley virus,SVV)replication.[Methods]IBRS-2 cells were infected with SVV,and the regulation of endogenous CTSS expression by SVV infection was investigated by RT-qPCR at transcriptional levels.The effect of SVV infection on CTSS enzyme activity was determined by ELISA.The regulatory effects of overexpressed CTSS on SVV replication and antiviral cytokines induced by SVV were detected by Western blotting(WB)and RT-qPCR.The specific siRNA for CTSS was synthesized and the interference effect of siRNA on CTSS and the effect of CTSS interference on SVV replication were detected by WB and RT-qPCR.[Results]The expression of endogenous CTSS and the activity of CTSS enzyme were markedly up-regulated in IBRS-2 cells infected with SVV.SVV replication in IBRS-2 cells was significantly inhibited and the expression of host antiviral cytokines host antiviral cytokines was up-regulated by CTSS overexpression.siRNA-2947 down-regulate the expression of endogenous CTSS to promote SVV replication.[Conclusion]CTSS inhibits SVV replication by enhancing the up-regulated expression of host antiviral cytokines.This study provided the reference basis for further exploring the role and mechanisms of host CTSS in anti-SVV innate immune response.