title: Analysis of murine B-cell epitopes on Eastern equine encephalitis virus glycoprotein E2.
abstract: The Eastern equine encephalitis virus (EEEV) E2 protein is one of the main targets of the protective immune response against EEEV. Although some efforts have done to elaborate the structure and immune molecular basis of Alphaviruses E2 protein, the published data of EEEV E2 are limited. Preparation of EEEV E2 protein-specific antibodies and define MAbs-binding epitopes on E2 protein will be conductive to the antibody-based prophylactic and therapeutic and to the study on structure and function of EEEV E2 protein. In this study, 51 EEEV E2 protein-reactive monoclonal antibodies (MAbs) and antisera (polyclonal antibodies, PAbs) were prepared and characterized. By pepscan with MAbs and PAbs using enzyme-linked immunosorbent assay, we defined 18 murine linear B-cell epitopes. Seven peptide epitopes were recognized by both MAbs and PAbs, nine epitopes were only recognized by PAbs, and two epitopes were only recognized by MAbs. Among the epitopes recognized by MAbs, seven epitopes were found only in EEEV and two epitopes were found both in EEEV and Venezuelan equine encephalitis virus (VEEV). Four of the EEEV antigenic complex-specific epitopes were commonly held by EEEV subtypes I/II/III/IV (1-16aa, 248-259aa, 271-286aa, 321-336aa probably located in E2 domain A, domain B, domain C, domain C, respectively). The remaining three epitopes were EEEV type-specific epitopes: a subtype I-specific epitope at amino acids 108-119 (domain A), a subtype I/IV-specific epitope at amino acids 211-226 (domain B) and a subtype I/II/III-specific epitope at amino acids 231-246 (domain B). The two common epitopes of EEEV and VEEV were located at amino acids 131-146 and 241-256 (domain B). The generation of EEEV E2-specific MAbs with defined specificities and binding epitopes will inform the development of differential diagnostic approaches and structure study for EEEV and associated alphaviruses.
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin 150001, People's Republic of China.
authors.forename: Sun
authors.initials: S
authors.lastname: EnCheng
authors.forename: Zhao
authors.initials: Z
authors.lastname: Jing
authors.forename: Yang
authors.initials: Y
authors.lastname: Tao
authors.forename: Xu
authors.initials: X
authors.lastname: QingYuan
authors.forename: Qin
authors.initials: Q
authors.lastname: Yongli
authors.forename: Wang
authors.initials: W
authors.lastname: WenShi
authors.forename: Wei
authors.initials: W
authors.lastname: Peng
authors.forename: Sun
authors.initials: S
authors.lastname: Liang
authors.forename: Sun
authors.initials: S
authors.lastname: Jing
authors.forename: Wu
authors.initials: W
authors.lastname: DongLai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D004663:Encephalitis Virus, Eastern Equine / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000276:immunology*; D004666:Encephalitis Virus, Venezuelan Equine / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000276:immunology; D004683:Encephalomyelitis, Equine / Q000276:immunology / Q000821:virology*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D051379:Mice; D013045:Species Specificity; D018411:Spodoptera; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D018985:Epitopes, B-Lymphocyte; D014759:Viral Envelope Proteins; C064956:glycoprotein E2, equine encephalitis virus