title: Identification of novel rabbit hemorrhagic disease virus B-cell epitopes and their interaction with host histo-blood group antigens.
abstract: Rabbit haemorrhagic disease, caused by rabbit hemorrhagic disease virus (RHDV), results in the death of millions of adult rabbits worldwide, with a mortality rate that exceeds 90%. The sole capsid protein, VP60, is divided into shell (S) and protruding (P) domains, and the more exposed P domain likely contains determinants for cell attachment and antigenic diversity. Nine mAbs against VP60 were screened and identified. To map antigenic epitopes, a set of partially overlapping and consecutive truncated proteins spanning VP60 were expressed. The minimal determinants of the linear B-cell epitopes of VP60 in the P domain, N(326)PISQV(331), D(338)MSFV(342) and K(562)STLVFNL(569), were recognized by one (5H3), four (1B8, 3D11, 4C2 and 4G2) and four mAbs (1D4, 3F7, 5G2 and 6B2), respectively. Sequence alignment showed epitope D(338)MSFV(342) was conserved among all RHDV isolates. Epitopes N(326)PISQV(331) and K(562)STLVFNL(569) were highly conserved among RHDV G1-G6 and variable in RHDV2 strains. Previous studies demonstrated that native viral particles and virus-like particles (VLPs) of RHDV specifically bound to synthetic blood group H type 2 oligosaccharides. We established an oligosaccharide-based assay to analyse the binding of VP60 and epitopes to histo-blood group antigens (HBGAs). Results showed VP60 and its epitopes (aa 326-331 and 338-342) in the P2 subdomain could significantly bind to blood group H type 2. Furthermore, mAbs 1B8 and 5H3 could block RHDV VLP binding to synthetic H type 2. Collectively, these two epitopes might play a key role in the antigenic structure of VP60 and interaction of RHDV and HBGA.
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Song
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Zhiyu
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Hu
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Liu
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Houjun
authors.initials: H
authors.lastname: Wei
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Jiabin
authors.initials: J
authors.lastname: Xue
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Weizhong
authors.initials: W
authors.lastname: Xu
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
authors.forename: Rulong
authors.initials: R
authors.lastname: Qiu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D001789:Blood Group Antigens / Q000378:metabolism*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology* / Q000378:metabolism*; D005260:Female; D017929:Hemorrhagic Disease Virus, Rabbit / Q000276:immunology*; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011817:Rabbits; D015678:Viral Structural Proteins / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D001789:Blood Group Antigens; D018985:Epitopes, B-Lymphocyte; D015678:Viral Structural Proteins; C081149:viral protein 60, rabbit hemorrhagic disease virus