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轮状病毒是引起婴幼儿腹泻的主要病原,VP4是RV重要的抗原蛋白,在早期病毒与细胞黏附过程中发挥重要作用,包括受体结合和细胞渗透。在细胞黏附过程中,VP4易被切割成VP5*和VP8*2个片段并以此增强病毒感染性。为了深入研究VP5*和VP8*的免疫学性质,进一步评价其应用前景,从TB-Chen株RV基因组中编码VP4蛋白基因上克隆了VP5*和VP8*开放读码框核苷酸序列,构建了表达质粒,在原核大肠杆菌系统中重组表达了VP5*和VP8*蛋白,进一步分析了它们的免疫学性质。结果显示,VP5*和VP8*可在E.coli中高效表达,重组蛋白VP5*(rVP5*)和VP8*(rVP8*)可诱导免疫豚鼠产生特异性血清抗体,这些抗体可特异性识别自身蛋白(rVP5*或rVP8*),可识别来自TB-Chen株的重组VP4蛋白,并可识别SA11和Wa感染的MA104细胞中合成的病毒VP4蛋白。这些结果表明,rVP5*和rVP8*蛋白具有较高的免疫原性,抗rVP5*和抗rVP8*的抗体具有高度特异性和交叉反应性。
Rotavirus is the leading cause of diarrhea in infants and young children. VP4 is an important antigenic protein of RV and plays an important role in early virus and cell adhesion process, including receptor binding and cell infiltration. During cell adhesion, VP4 is easily cleaved into VP5 * and VP8 * 2 fragments and thus enhances viral infectivity. In order to further study the immunological properties of VP5 * and VP8 *, and to further evaluate their application prospects, VP5 * and VP8 * open reading frame nucleotide sequences were cloned from the VP4 protein gene of the RV genome of TB-Chen strain Expression plasmids. VP5 * and VP8 * proteins were recombinantly expressed in prokaryotic E.coli, and their immunological properties were further analyzed. The results showed that VP5 * and VP8 * can be highly expressed in E.coli. Recombinant proteins VP5 * (rVP5 *) and VP8 * (rVP8 *) can induce immune guinea pigs to produce specific serum antibodies. These antibodies can specifically recognize self-proteins (rVP5 * or rVP8 *) recognizes the recombinant VP4 protein from the TB-Chen strain and recognizes the synthetic viral VP4 protein in SAlO and Wa infected MA104 cells. These results indicate that the rVP5 * and rVP8 * proteins are highly immunogenic and the anti-rVP5 * and anti-rVP8 * antibodies are highly specific and cross-reactive.