Antigenic analysis of grass carp reovirus using single-chain variable fragment antibody against IgM from Ctenopharyngodon idella

Abstract

Grass carp (Ctenopharyngodon idella)is an important species of freshwater aquaculture fish in China. However, grass carp reovirus (GCRV) can cause fatal hemorrhagic disease in yearling populations. Until now, a strategy to define the antigenic capacity of the virus's structural proteins for preparing an effective vaccine has not been available. In this study, some single-chain variable fragment antibodies (scFv), which could specifically recognize grass carp IgM, were selected from a constructed mouse naïve antibody phage display cDNA library. The identified scFv C1B3 clone was shown to possess relatively higher specific binding activity to grass carp IgM. Furthermore, ELISA analysis indicated that the IgM level in serum from virus-infected grass carp was more than two times higher than that of the control group at 5-7 days post infection. Moreover, Western blot analysis demonstrated that the outer capsid protein VP7 has a specific immuno-binding-reaction with the serum IgM from virus-infected grass carp. Our results suggest that VP7 can induce a stronger immune response in grass carp than the other GCRV structural proteins, which implies that VP7 protein could be used as a preferred immunogen for vaccine design.

Large-scale SNP screenings identify markers linked with GCRV resistant traits through transcriptomes of individuals and cell lines in Ctenopharyngodon idella

“…Interestingly, D. rerio post GCRV challenge is nearly 100% survival 20 . In order to investigate the differences among them, comparisons of genomic base types of the corresponding SNP positions among C. idella , G. rarus and D. rerio were listed in Supplementary Table S2.…”

Yeast Surface Display of Capsid Protein VP7 of Grass Carp Reovirus: Fundamental Investigation for the Development of Vaccine Against Hemorrhagic Disease

“…The antiserum generated from grass carps infected with GCRV could only recognize outer capsid protein VP7 of GCRV, which indicates that VP7 is an essential antigenic protein of GCRV [8]. In another report, both antisera generated from GCRV capsid proteins VP5 and VP7, respectively, showed neutralization ability in GCRV-infected CIK (Ctenopharyngodon idellus kidney) cells.…”

The distribution of different virulence grass carp reovirus strains in some neglected tissues

“…Currently, no effective antiviral treatment against GCRV infection is available (Ma et al 2011). In an attempt to control the spread of the disease, several inactive vaccines have been developed over the years, yet epidemic outbreaks have still occurred in many freshwater areas in recent years (Chen et al 2013). Therefore, preventing this infectious disease still remains a great challenge (Cheng et al 2008) and requires a better understanding of the pathogenic mechanisms underpinning the development of GCRV driven hemorrhagic disease.…”