Department of Comparative Physiology, Uppsala University, Norbyvägen 18A, SE-75236 Uppsala, Sweden.
J Virol. 2010 Oct;84(20):10844-51. doi: 10.1128/JVI.01045-10. Epub 2010 Aug 4.
The gC1qR/p32 protein is a multiple receptor for several proteins and pathogens. We cloned a gC1qR homologue in a crustacean, Pacifastacus leniusculus, and analyzed the expression of P. leniusculus C1qR (PlgC1qR) in various tissues. The gC1qR/p32 transcript was significantly enhanced by white spot syndrome virus (WSSV) infection 6 h after viral infection both in vitro in a hematopoietic tissue cell culture (Hpt) and in vivo compared to appropriate controls. Moreover, PlgC1qR silencing in both the Hpt cell culture and live crayfish enhanced the WSSV replication. In addition, by making a recombinant PlgC1qR protein we could show that if this recombinant protein was injected in a crayfish, Pacifastacus leniusculus, followed by injection of WSSV, this significantly reduced viral replication in vivo. Furthermore, if the recombinant PlgC1qR was incubated with Hpt cells and then WSSV was added, this also reduced viral replication. These experiments clearly demonstrate that recombinant PlgC1qR reduce WSSV replication both in vivo and in vitro. The results from a far-Western overlay and glutathione S-transferase pull-down assays showed that PlgC1qR could bind to VP15, VP26, and VP28. Altogether, these results demonstrate a role for PlgC1qR in antiviral activity against WSSV.
gC1qR/p32 蛋白是多种蛋白和病原体的多受体。我们在甲壳动物太平洋螯虾(Pacifastacus leniusculus)中克隆了 gC1qR 同源物,并分析了 P. leniusculus C1qR(PlgC1qR)在各种组织中的表达。与适当的对照相比,gC1qR/p32 转录物在体外造血组织细胞培养(Hpt)和体内在感染病毒后 6 小时均被白斑综合征病毒(WSSV)感染显著增强。此外,在 Hpt 细胞培养和活体螯虾中沉默 PlgC1qR 均增强了 WSSV 的复制。此外,通过制备重组 PlgC1qR 蛋白,我们可以证明如果将这种重组蛋白注射到螯虾中,随后再注射 WSSV,这将显著减少体内的病毒复制。此外,如果将重组 PlgC1qR 与 Hpt 细胞孵育,然后再加入 WSSV,这也会减少病毒复制。这些实验清楚地表明,重组 PlgC1qR 可减少体内和体外的 WSSV 复制。远 Western 印迹和谷胱甘肽 S-转移酶下拉测定的结果表明,PlgC1qR 可以与 VP15、VP26 和 VP28 结合。总之,这些结果表明 PlgC1qR 在抗病毒活性中对 WSSV 具有作用。