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Development of a novel candidate subunit vaccine against Grass carp reovirus Guangdong strain (GCRV-GD108)

文献类型： 外文期刊

作者： Tian, Yuanyuan 1 ; Ye, Xing 1 ; Zhang, Lili 1 ; Deng, Guocheng 1 ; Bai, Yueqiang 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst,Minist Agr, Lab Aquat Anim Genet Engn & Mol Breeding, Key Lab Trop & Subtrop Fisheries Resource Utiliza, Guangzhou 510380, Peoples R China

关键词： VP4;Outer capsid protein;B cell epitopes;Neutralization assay;Relative percent survival

期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.581； 五年影响因子：4.851 ）

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收录情况： SCI

摘要： Grass carp reovirus Guangdong 108 strain (GCRV-GD108) was recently isolated in Guangdong province, China. M6 gene of GCRV-GD108 was speculated encoding virus major outer capsid protein VP4. Blast analysis showed that the amino acid sequence of GCRV-GD108 VP4 was homologous to the structural protein VP4 of known Aquareoviruses (27.3-32.9%). Immunogenicity prediction by DNAStar software revealed there were multiple B cell epitopes on GCRV-GD108 VP4. Prokaryotic expression vector pET32a was used to express VP4 recombinant protein (rVP4) in E. coli BL21(DE3) strain. As expected, the molecular weight of recombinant VP4 was about 87 kDa showed by SOS-PAGE result. Neutralization assay demonstrated that the rabbit polyclonal antibody of rVP4 could prevent virus infection efficiently. After 14 days immunization with the rVP4, grass carps were challenged with GCRV-GD108, the results showed that different doses of rVP4 (1 mu g/g, 3 mu g/g and 5 mu g/g) all provided protection against virus infection (47-82%). The relative percent survival reached 82% in the group immunized with 3 mu g/g of rVP4. ELISA revealed rVP4 induced high antibody titer in immunized fish. IgM expression levels in head kidney of grass carp were detected by RT-PCR, and the results showed that IgM expressed at a significantly higher level in immunization groups than in blank control, indicating the rVP4 can induce strong immune response. In conclusion, rVP4 is a candidate vaccine against GCRV-GD108

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