Department of Genetics, University of Málaga, 29071 Málaga, Spain.
Mol Immunol. 2011 Oct;49(1-2):107-14. doi: 10.1016/j.molimm.2011.07.023. Epub 2011 Aug 27.
Mx proteins are crucial effectors of the innate antiviral response mediated by the interferon type I signalling pathway. Recently, three Mx proteins, named SauMx1, SauMx2 and SauMx3, corresponding to three different genes, have been identified in the cultured marine species gilthead seabream (Sparus aurata). In this study, the three SauMx cDNAs were cloned into expression vectors and used for transfection of CHSE-214 cells. Monoclonal cell populations stably expressing each recombinant protein have been obtained and characterized. The protection conferred by each recombinant SauMx against Infectious Pancreatic Necrosis Virus (IPNV) infection has been in vitro evaluated, having found clear differences among them. According to the cytopathic effects and the virus yield reduction assays, only cells expressing SauMx2 and SauMx3 showed significant resistance to IPNV infection. Otherwise, quantitative RT real-time PCR assays suggested that each SauMx protein has a different target during the viral inhibition process. The differences observed among the three SauMx proteins are discussed in terms of their differential mechanism of action and antiviral specificity, suggesting, as a whole, to play a synergistic activity in the protection of gilthead seabream against IPNV.
Mx 蛋白是干扰素 I 信号通路介导的先天抗病毒反应的关键效应因子。最近,在养殖的海洋物种真鲷(Sparus aurata)中鉴定出三种 Mx 蛋白,分别命名为 SauMx1、SauMx2 和 SauMx3,它们对应三个不同的基因。在本研究中,将这三种 SauMx cDNA 克隆到表达载体中,并用于转染 CHSE-214 细胞。获得了稳定表达每种重组蛋白的单克隆细胞群,并对其进行了表征。体外评估了每种重组 SauMx 对传染性胰腺坏死病毒(IPNV)感染的保护作用,发现它们之间存在明显差异。根据细胞病变效应和病毒产量减少测定,只有表达 SauMx2 和 SauMx3 的细胞对 IPNV 感染表现出显著的抗性。然而,定量 RT 实时 PCR 检测表明,每种 SauMx 蛋白在病毒抑制过程中都有不同的靶标。本文讨论了这三种 SauMx 蛋白之间的差异,涉及它们的作用机制和抗病毒特异性的差异,表明它们在保护真鲷抵抗 IPNV 方面整体发挥协同作用。