title: Comprehensive analysis of varicella-zoster virus proteins using a new monoclonal antibody collection.
abstract: Varicella-zoster virus (VZV) is the etiological agent of chickenpox and shingles. Due to the virus's restricted host and cell type tropism and the lack of tools for VZV proteomics, it is one of the least-characterized human herpesviruses. We generated 251 monoclonal antibodies (MAbs) against 59 of the 71 (83%) currently known unique VZV proteins to characterize VZV protein expression in vitro and in situ. Using this new set of MAbs, 44 viral proteins were detected by Western blotting (WB) and indirect immunofluorescence (IF); 13 were detected by WB only, and 2 were detected by IF only. A large proportion of viral proteins was analyzed for the first time in the context of virus infection. Our study revealed the subcellular localization of 46 proteins, 14 of which were analyzed in detail by confocal microscopy. Seven viral proteins were analyzed in time course experiments and showed a cascade-like temporal gene expression pattern similar to those of other herpesviruses. Furthermore, selected MAbs tested positive on human skin lesions by using immunohistochemistry, demonstrating the wide applicability of the MAb collection. Finally, a significant portion of the VZV-specific antibodies reacted with orthologs of simian varicella virus (SVV), thus enabling the systematic analysis of varicella in a nonhuman primate model system. In summary, this study provides insight into the potential function of numerous VZV proteins and novel tools to systematically study VZV and SVV pathogenesis.
authors.affiliation: Center for Proteomics, Faculty of Medicine, University of Rijeka, Rijeka, Croatia.
authors.forename: Tihana
authors.initials: T
authors.lastname: Lenac Roviš
authors.forename: Susanne M
authors.initials: SM
authors.lastname: Bailer
authors.forename: Venkata R
authors.initials: VR
authors.lastname: Pothineni
authors.forename: Werner J D
authors.initials: WJ
authors.lastname: Ouwendijk
authors.forename: Hrvoje
authors.initials: H
authors.lastname: Šimić
authors.forename: Marina
authors.initials: M
authors.lastname: Babić
authors.forename: Karmela
authors.initials: K
authors.lastname: Miklić
authors.forename: Suzana
authors.initials: S
authors.lastname: Malić
authors.forename: Marieke C
authors.initials: MC
authors.lastname: Verweij
authors.forename: Armin
authors.initials: A
authors.lastname: Baiker
authors.forename: Orland
authors.initials: O
authors.lastname: Gonzalez
authors.forename: Albrecht
authors.initials: A
authors.lastname: von Brunn
authors.forename: Ralf
authors.initials: R
authors.lastname: Zimmer
authors.forename: Klaus
authors.initials: K
authors.lastname: Früh
authors.forename: Georges M G M
authors.initials: GM
authors.lastname: Verjans
authors.forename: Stipan
authors.initials: S
authors.lastname: Jonjić
authors.forename: Jürgen
authors.initials: J
authors.lastname: Haas
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D002644:Chickenpox / Q000821:virology; D004847:Epithelial Cells / Q000821:virology; D019084:Fluorescent Antibody Technique, Indirect; D006562:Herpes Zoster / Q000821:virology; D014645:Herpesvirus 3, Human / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D040901:Proteomics; D012867:Skin / Q000276:immunology / Q000821:virology; D014764:Viral Proteins / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D014764:Viral Proteins