title: Epitope mapping of human herpesvirus-7 gp65 using monoclonal antibodies.
abstract: Human herpesvirus (HHV)-7 encodes a unique 65-kDa heparin-binding glycoprotein, designated gp65. This molecule is thought to play a role in virus attachment and entry. To obtain reagents to map the structure and function of HHV-7 gp65, we produced monoclonal antibodies to this molecule. Ten monoclonal antibodies reacting with gp65 on ELISA were subdivided in four groups on the basis of their isotype and differential reactivity with (i) native versus denatured forms of gp65, and (ii) mature (virion-associated) versus immature (cell-associated) forms of the molecule. We were able to map the binding epitopes for eight of these ten antibodies, and these were found to cluster to one site on gp65 (amino acids 239-278); within this region, the antibodies reacted with at least three distinct domains (244-251, 255-262, 263-278). The reasons for the apparent immunodominance of this region are uncertain. Taken together, this panel of antibodies constitutes an extensive and well-characterized set of HHV-7 specific antibodies that may have utility for future analyses of the structure/function of gp65, and for studies on the virus life cycle.
authors.affiliation: Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York 14642, USA.
authors.forename: D
authors.initials: D
authors.lastname: Skrincosky
authors.forename: R A
authors.initials: RA
authors.lastname: Willis
authors.forename: P K
authors.initials: PK
authors.lastname: Hocknell
authors.forename: J G
authors.initials: JG
authors.lastname: Frelinger
authors.forename: P
authors.initials: P
authors.lastname: Mirandola
authors.forename: X
authors.initials: X
authors.lastname: Wang
authors.forename: S
authors.initials: S
authors.lastname: Dewhurst
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology*; D016199:Herpesvirus 7, Human / Q000276:immunology*; D006801:Humans; D007132:Immunoglobulin Isotypes; D051379:Mice; D008969:Molecular Sequence Data; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; C407562:GP65 protein, Human herpesvirus 7; D006023:Glycoproteins; D007132:Immunoglobulin Isotypes; D014759:Viral Envelope Proteins