title: The major neutralizing antigenic site on herpes simplex virus glycoprotein D overlaps a receptor-binding domain.
abstract: Herpes simplex virus (HSV) entry is dependent on the interaction of virion glycoprotein D (gD) with one of several cellular receptors. We previously showed that gD binds specifically to two structurally dissimilar receptors, HveA and HveC. We have continued our studies by using (i) a panel of baculovirus-produced gD molecules with various C-terminal truncations and (ii) a series of gD mutants with nonoverlapping 3-amino-acid deletions between residues 222 and 254. Binding of the potent neutralizing monoclonal antibody (MAb) DL11 (group Ib) was unaffected in forms of gD containing residues 1 to 250 but was greatly diminished in molecules truncated at residue 240 or 234. Both receptor binding and blocking of HSV infection were also affected by these C-terminal truncations. gD-1(234t) bound weakly to both HveA and HveC as determined by enzyme-linked immunosorbent assay (ELISA) and failed to block infection. Interestingly, gD-1(240t) bound well to both receptors but blocked infection poorly, indicating that receptor binding as measured by ELISA is not the only gD function required for blocking. Optical biosensor studies showed that while gD-1(240t) bound HveC with an affinity similar to that of gD-1(306t), the rates of complex formation and dissociation were significantly faster than for gD-1(306t). Complementation analysis showed that any 3-amino-acid deletion between residues 222 and 251 of gD resulted in a nonfunctional protein. Among this set of proteins, three had lost DL11 reactivity (those with deletions between residues 222 and 230). One of these proteins (deletion 222-224) was expressed as a soluble form in the baculovirus system. This protein did not react with DL11, bound to both HveA and HveC poorly as shown by ELISA, and failed to block HSV infection. Since this protein was bound by several other MAbs that recognize discontinuous epitopes, we conclude that residues 222 to 224 are critical for gD function. We propose that the potent virus-neutralizing activity of DL11 (and other group Ib MAbs) likely reflects an overlap between its epitope and a receptor-binding domain of gD.
authors.affiliation: School of Dental Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. whitbeck@biochem.dental.upenn.edu
authors.forename: J C
authors.initials: JC
authors.lastname: Whitbeck
authors.forename: M I
authors.initials: MI
authors.lastname: Muggeridge
authors.forename: A H
authors.initials: AH
authors.lastname: Rux
authors.forename: W
authors.initials: W
authors.lastname: Hou
authors.forename: C
authors.initials: C
authors.lastname: Krummenacher
authors.forename: H
authors.initials: H
authors.lastname: Lou
authors.forename: A
authors.initials: A
authors.lastname: van Geelen
authors.forename: R J
authors.initials: RJ
authors.lastname: Eisenberg
authors.forename: G H
authors.initials: GH
authors.lastname: Cohen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D016367:Baculoviridae; D001665:Binding Sites; D015374:Biosensing Techniques; D002460:Cell Line; D002522:Chlorocebus aethiops; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D015870:Gene Expression; D005807:Genes, Overlapping; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009500:Neutralization Tests; D018124:Receptors, Tumor Necrosis Factor; D053240:Receptors, Tumor Necrosis Factor, Member 14; D011991:Receptors, Virus / Q000378:metabolism*; D017384:Sequence Deletion; D012995:Solubility; D018411:Spodoptera / Q000166:cytology; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000956:Antigens, Viral; D018985:Epitopes, B-Lymphocyte; D018124:Receptors, Tumor Necrosis Factor; D053240:Receptors, Tumor Necrosis Factor, Member 14; D011991:Receptors, Virus; C506028:TNFRSF14 protein, human; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1