title: Functional screening for anti-CMV biologics identifies a broadly neutralizing epitope of an essential envelope protein.
abstract: The prototypic β-herpesvirus human cytomegalovirus (CMV) establishes life-long persistence within its human host. The CMV envelope consists of various protein complexes that enable wide viral tropism. More specifically, the glycoprotein complex gH/gL/gO (gH-trimer) is required for infection of all cell types, while the gH/gL/UL128/130/131a (gH-pentamer) complex imparts specificity in infecting epithelial, endothelial and myeloid cells. Here we utilize state-of-the-art robotics and a high-throughput neutralization assay to screen and identify monoclonal antibodies (mAbs) targeting the gH glycoproteins that display broad-spectrum properties to inhibit virus infection and dissemination. Subsequent biochemical characterization reveals that the mAbs bind to gH-trimer and gH-pentamer complexes and identify the antibodies' epitope as an 'antigenic hot spot' critical for virus entry. The mAbs inhibit CMV infection at a post-attachment step by interacting with a highly conserved central alpha helix-rich domain. The platform described here provides the framework for development of effective CMV biologics and vaccine design strategies.
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Gardner
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Kathryn R
authors.initials: KR
authors.lastname: Stein
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.|Center for Therapeutic Antibody Development, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: J Andrew
authors.initials: JA
authors.lastname: Duty
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Toni M
authors.initials: TM
authors.lastname: Schwarz
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Vanessa M
authors.initials: VM
authors.lastname: Noriega
authors.affiliation: Center for Therapeutic Antibody Development, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kraus
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.|Center for Therapeutic Antibody Development, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Moran
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Domenico
authors.initials: D
authors.lastname: Tortorella
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000627:therapeutic use; D057134:Antibodies, Neutralizing / Q000737:chemistry / Q000627:therapeutic use*; D002460:Cell Line; D003587:Cytomegalovirus / Q000472:pathogenicity*; D003586:Cytomegalovirus Infections / Q000276:immunology / Q000517:prevention & control*; D006801:Humans; D051379:Mice; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology*; D014765:Viral Vaccines; D053586:Virus Internalization
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C071440:glycoprotein H, Cytomegalovirus