P[3]rotavirus(RV)has been identified in many species,including human,simian,dog,and bat.Several glycans,including sialic acid,histo-blood group antigens(HBGAs)are reported as RV attachment factors.The glycan binding s...P[3]rotavirus(RV)has been identified in many species,including human,simian,dog,and bat.Several glycans,including sialic acid,histo-blood group antigens(HBGAs)are reported as RV attachment factors.The glycan binding specificity of different P[3]RV VP8*s were investigated in this study.Human HCR3 A and dog P[3]RV VP8*s recognized glycans with terminal sialic acid and hemagglutinated the red blood cells,while bat P[3]VP8*showed neither binding to glycans nor hemagglutination.However,the bat P[3]VP8*mutant of C189 Y obtained the ability to hemagglutinate the red blood cells,while human P[3]HCR3 A/M2-102 mutants of Y189 C lost the ability.Sequence alignment and structural analysis indicated that residue 189 played an important role in the ligand recognition and may contribute to the cross-species transmission.Structural superimposition exhibited that bat P[3]VP8*model was quite different from the simian P[3]Rhesus rotavirus(RRV)P[3]VP8*,indicating that bat P[3]RV was relatively distinct and partially contributed to the no binding to tested glycans.These results promote our understanding of P[3]VP8*/glycans interactions and the potential transmission of bat/human P[3]RVs,offering more insight into the RV infection and prevalence.展开更多
轮状病毒(Rotaviruses,RVs)是引起人和动物病毒性腹泻的重要病原,本文旨在研究猪C组轮状病毒(Group C rotaviruses,RVCs)VP8*蛋白的受体结合特征。本研究合成一株P[5]型猪C组轮状病毒(Por2011)的VP8*基因,经体外原核表达,采用亲和层析...轮状病毒(Rotaviruses,RVs)是引起人和动物病毒性腹泻的重要病原,本文旨在研究猪C组轮状病毒(Group C rotaviruses,RVCs)VP8*蛋白的受体结合特征。本研究合成一株P[5]型猪C组轮状病毒(Por2011)的VP8*基因,经体外原核表达,采用亲和层析纯化获得VP8*目的蛋白,利用糖点阵实验、寡糖结合实验研究其寡糖结合特征,再通过序列比对和突变分析确定其潜在受体结合位点。结果显示猪C组轮状病毒Por2011 VP8*蛋白与P1抗原特异性结合,但第108位氨基酸突变后的蛋白则不能与P1抗原结合。本研究表明猪C组轮状病毒的某些型别可能以P1抗原为潜在受体,该受体结合位点与人C组轮状病毒的受体结合位点较为接近,这为猪C组轮状病毒感染机制的研究提供了一定的依据和基础。展开更多
Lumazine synthase from Brucella spp. (BLS) is a highly immunogenic decameric protein. It has been previously described as a carrier of peptides or proteins to increase their immunogenicity in different animal species,...Lumazine synthase from Brucella spp. (BLS) is a highly immunogenic decameric protein. It has been previously described as a carrier of peptides or proteins to increase their immunogenicity in different animal species, but its activity has never been evaluated in chickens. In this work, the use of BLS to improve the antibody response against bovine rotavirus (BRV) VP8d protein in laying hens was assessed. VP8d is the inner domain of the VP8 spike protein which preserves the sialic acid binding activity and the neutralizing epitopes present in the viral protein. Hens were immunized three times with 2 μg of VP8d alone or fused to BLS. Hens inoculated with BLSVP8d developed higher antibody titers (evaluated by ELISA and viral neutralization test) than hens immunized either with VP8d alone or the mixture of VP8d and BLS. Furthermore, IgY antibodies against BLSVP8d were able to fully protect mice against challenge with virulent BRV in a dose-depent-manner. Overall, these results demonstrate that BLS is a potent immonumodulator that enhances the antibody response in hens, thus increasing the concentration of specific IgY in the egg yolk, one of the main issues to be adressed in order to improve the use of the IgY technology.展开更多
基金the participation of the ProteinGlycan Interaction Resource of the CFG(supporting grant R24 GM098791)the National Center for Functional Glycomics(NCFG)at Beth Israel Deaconess Medical Center,Harvard Medical School(supporting grant P41 GM103694)+1 种基金supported by grants from the National Natural Science Foundation of China(NSFC)(No.21934005)National Key Research and Development Program of China(2018YFC1200602)。
文摘P[3]rotavirus(RV)has been identified in many species,including human,simian,dog,and bat.Several glycans,including sialic acid,histo-blood group antigens(HBGAs)are reported as RV attachment factors.The glycan binding specificity of different P[3]RV VP8*s were investigated in this study.Human HCR3 A and dog P[3]RV VP8*s recognized glycans with terminal sialic acid and hemagglutinated the red blood cells,while bat P[3]VP8*showed neither binding to glycans nor hemagglutination.However,the bat P[3]VP8*mutant of C189 Y obtained the ability to hemagglutinate the red blood cells,while human P[3]HCR3 A/M2-102 mutants of Y189 C lost the ability.Sequence alignment and structural analysis indicated that residue 189 played an important role in the ligand recognition and may contribute to the cross-species transmission.Structural superimposition exhibited that bat P[3]VP8*model was quite different from the simian P[3]Rhesus rotavirus(RRV)P[3]VP8*,indicating that bat P[3]RV was relatively distinct and partially contributed to the no binding to tested glycans.These results promote our understanding of P[3]VP8*/glycans interactions and the potential transmission of bat/human P[3]RVs,offering more insight into the RV infection and prevalence.
文摘轮状病毒(Rotaviruses,RVs)是引起人和动物病毒性腹泻的重要病原,本文旨在研究猪C组轮状病毒(Group C rotaviruses,RVCs)VP8*蛋白的受体结合特征。本研究合成一株P[5]型猪C组轮状病毒(Por2011)的VP8*基因,经体外原核表达,采用亲和层析纯化获得VP8*目的蛋白,利用糖点阵实验、寡糖结合实验研究其寡糖结合特征,再通过序列比对和突变分析确定其潜在受体结合位点。结果显示猪C组轮状病毒Por2011 VP8*蛋白与P1抗原特异性结合,但第108位氨基酸突变后的蛋白则不能与P1抗原结合。本研究表明猪C组轮状病毒的某些型别可能以P1抗原为潜在受体,该受体结合位点与人C组轮状病毒的受体结合位点较为接近,这为猪C组轮状病毒感染机制的研究提供了一定的依据和基础。
文摘Lumazine synthase from Brucella spp. (BLS) is a highly immunogenic decameric protein. It has been previously described as a carrier of peptides or proteins to increase their immunogenicity in different animal species, but its activity has never been evaluated in chickens. In this work, the use of BLS to improve the antibody response against bovine rotavirus (BRV) VP8d protein in laying hens was assessed. VP8d is the inner domain of the VP8 spike protein which preserves the sialic acid binding activity and the neutralizing epitopes present in the viral protein. Hens were immunized three times with 2 μg of VP8d alone or fused to BLS. Hens inoculated with BLSVP8d developed higher antibody titers (evaluated by ELISA and viral neutralization test) than hens immunized either with VP8d alone or the mixture of VP8d and BLS. Furthermore, IgY antibodies against BLSVP8d were able to fully protect mice against challenge with virulent BRV in a dose-depent-manner. Overall, these results demonstrate that BLS is a potent immonumodulator that enhances the antibody response in hens, thus increasing the concentration of specific IgY in the egg yolk, one of the main issues to be adressed in order to improve the use of the IgY technology.
