Grass carp reovirus(GCRV)is the causative agent of grass carp hemorrhagic disease,which has detrimental effects on the grass carp aquaculture industry.There are four known genotypes of GCRV and strains of different GC...Grass carp reovirus(GCRV)is the causative agent of grass carp hemorrhagic disease,which has detrimental effects on the grass carp aquaculture industry.There are four known genotypes of GCRV and strains of different GCRV genotypes differ greatly.In this study,the diversity of the protein VP6 from different GCRV stains and the effect of genotype GCRV 096 on replication was investigated in CIK cells.Our results showed that the VP6 protein of GCRV 096(genotypeІ)exhibited limited homology to that of GCRV GD108(genotypeШ),with few residues conserved in predicted protein-protein interaction domains.GCRV 096 VP6 protein was expressed and purified and an antiserum against it was characterized.Addition of purified VP6 protein or antiserum to culture media of CIK cells inhibited the replication of GCRV 096 in these cells.In contrast,replication of GCRV GD108(genotypeШ)was not affected in CIK cells under the same condition.Overall,our results indicated that the protein VP6 and VP6 antiserum did not provide cross-protection against GCRV strains and this can be attributed to differences among GCRV genotypes.It will be important to consider multiple GCRV genotypes in the development of effective GCRV vaccines and other therapies against grass carp hemorrhagic disease.In addition,bioinformatics analysis also suggested that the protein VP6 may have a role in the process of GCRV infection.This study lays the foundation for the prevention of grass carp hemorrhagic disease and further detailed studies on the pathogenesis of GCRV.展开更多
Elevated expression of the rotavirus VP6 antigen in transgenic plants is a critical factor in the development of a safe and effective rotavirus vaccine. Using codon optimization, a gene that encodes the inner capsid p...Elevated expression of the rotavirus VP6 antigen in transgenic plants is a critical factor in the development of a safe and effective rotavirus vaccine. Using codon optimization, a gene that encodes the inner capsid protein VP6 of the human group A rotavirus was synthesized （sVP6）. The VP6 and sVp6 genes were transformed into tobacco （Nicotiana tabacum L.） plants using Agrobacterium tumefaciens. The expression level of the sVP6 gene in transgenic plants was 3.8-34-fold higher than that of controls containing the non-modified VP6 gene, accounting for up to 0.34% of the total soluble protein （TSP）. Then, BALB/ c female mice that had been gavaged weekly with 10 mg TSP containing 34 p.g VP6 protein, in which VP6-specific serum IgG and mucosal IgA antibodies were investigated. The severity and duration of diarrhea caused by simian rotavirus SA-11 challenge were reduced significantly in passively immunized pups, which indicates that anti-VP6 antibodies generated in orally immunized female mice can be passed onto pups and provide heterotypic protection. An edible vaccine based on the VP6 of human rotavirus group A could provide a means to protect children and young animals from severe acute diarrhea.展开更多
基金This study was supported by National Natural Science Foundation of China(No.31602199)the Natural Science Foundation of Guangdong Province in China(No.2015A030313622)+3 种基金the Foundation of Guangdong Ocean University(K15246)the Foundation of Zhanjiang(No.2015A03027)the National 973 Plan Project in China(No.2009CB118704)and the project of Department of Education of Guangdong province in China(No.2012LYM_0075).
文摘Grass carp reovirus(GCRV)is the causative agent of grass carp hemorrhagic disease,which has detrimental effects on the grass carp aquaculture industry.There are four known genotypes of GCRV and strains of different GCRV genotypes differ greatly.In this study,the diversity of the protein VP6 from different GCRV stains and the effect of genotype GCRV 096 on replication was investigated in CIK cells.Our results showed that the VP6 protein of GCRV 096(genotypeІ)exhibited limited homology to that of GCRV GD108(genotypeШ),with few residues conserved in predicted protein-protein interaction domains.GCRV 096 VP6 protein was expressed and purified and an antiserum against it was characterized.Addition of purified VP6 protein or antiserum to culture media of CIK cells inhibited the replication of GCRV 096 in these cells.In contrast,replication of GCRV GD108(genotypeШ)was not affected in CIK cells under the same condition.Overall,our results indicated that the protein VP6 and VP6 antiserum did not provide cross-protection against GCRV strains and this can be attributed to differences among GCRV genotypes.It will be important to consider multiple GCRV genotypes in the development of effective GCRV vaccines and other therapies against grass carp hemorrhagic disease.In addition,bioinformatics analysis also suggested that the protein VP6 may have a role in the process of GCRV infection.This study lays the foundation for the prevention of grass carp hemorrhagic disease and further detailed studies on the pathogenesis of GCRV.
文摘Elevated expression of the rotavirus VP6 antigen in transgenic plants is a critical factor in the development of a safe and effective rotavirus vaccine. Using codon optimization, a gene that encodes the inner capsid protein VP6 of the human group A rotavirus was synthesized （sVP6）. The VP6 and sVp6 genes were transformed into tobacco （Nicotiana tabacum L.） plants using Agrobacterium tumefaciens. The expression level of the sVP6 gene in transgenic plants was 3.8-34-fold higher than that of controls containing the non-modified VP6 gene, accounting for up to 0.34% of the total soluble protein （TSP）. Then, BALB/ c female mice that had been gavaged weekly with 10 mg TSP containing 34 p.g VP6 protein, in which VP6-specific serum IgG and mucosal IgA antibodies were investigated. The severity and duration of diarrhea caused by simian rotavirus SA-11 challenge were reduced significantly in passively immunized pups, which indicates that anti-VP6 antibodies generated in orally immunized female mice can be passed onto pups and provide heterotypic protection. An edible vaccine based on the VP6 of human rotavirus group A could provide a means to protect children and young animals from severe acute diarrhea.
