title: Major neutralizing sites on vaccinia virus glycoprotein B5 are exposed differently on variola virus ortholog B6.
abstract: Immunization against smallpox (variola virus) with Dryvax, a live vaccinia virus (VV), was effective, but now safety is a major concern. To overcome this issue, subunit vaccines composed of VV envelope proteins from both forms of infectious virions, including the extracellular enveloped virion (EV) protein B5, are being developed. However, since B5 has 23 amino acid differences compared with its B6 variola virus homologue, B6 might be a better choice for such a strategy. Therefore, we compared the properties of both proteins using a panel of monoclonal antibodies (MAbs) to B5 that we had previously characterized and grouped according to structural and functional properties. The B6 gene was obtained from the Centers for Disease Control and Prevention, and the ectodomain was cloned and expressed in baculovirus as previously done with B5, allowing us to compare the antigenic properties of the proteins. Polyclonal antibodies to B5 or B6 cross-reacted with the heterologous protein, and 16 of 26 anti-B5 MAbs cross-reacted with B6. Importantly, 10 anti-B5 MAbs did not cross-react with B6. Of these, three have important anti-VV biologic properties, including their ability to neutralize EV infectivity and block comet formation. Here, we found that one of these three MAbs protected mice from a lethal VV challenge by passive immunization. Thus, epitopes that are present on B5 but not on B6 would generate an antibody response that would not recognize B6. Assuming that B6 contains similar variola virus-specific epitopes, our data suggest that a subunit vaccine using the variola virus homologues might exhibit improved protective efficacy against smallpox.
authors.affiliation: Department of Microbiology, School of Dental Medicine, University of Pennsylvania, 240 S. 40th St., Philadelphia, PA 19104-6002, USA. lyriaaldaz@gmail.com
authors.forename: Lydia
authors.initials: L
authors.lastname: Aldaz-Carroll
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Xiao
authors.forename: J Charles
authors.initials: JC
authors.lastname: Whitbeck
authors.forename: Manuel Ponce
authors.initials: MP
authors.lastname: de Leon
authors.forename: Huan
authors.initials: H
authors.lastname: Lou
authors.forename: Mikyung
authors.initials: M
authors.lastname: Kim
authors.forename: Jessica
authors.initials: J
authors.lastname: Yu
authors.forename: Ellis L
authors.initials: EL
authors.lastname: Reinherz
authors.forename: Stuart N
authors.initials: SN
authors.lastname: Isaacs
authors.forename: Roselyn J
authors.initials: RJ
authors.lastname: Eisenberg
authors.forename: Gary H
authors.initials: GH
authors.lastname: Cohen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D020552:Comet Assay; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011817:Rabbits; D012900:Smallpox Vaccine / Q000276:immunology*; D014616:Vaccinia virus / Q000276:immunology*; D012901:Variola virus / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D012900:Smallpox Vaccine; D014759:Viral Envelope Proteins