Antigenic and functional analysis of a neutralization site of HSV-1 glycoprotein D.
Herpes simplex virus glycoprotein D is a component of the virion envelope and appears to be involved in attachment, penetration, and cell fusion. Monoclonal antibodies (MAbs) against this protein can be arranged in groups, on the basis of a number of biological and biochemical properties. Group I antibodies are type-common, have high complement-independent neutralization titers, recognize discontinuous (conformational) epitopes, and block each other in a binding assay. The sum of their epitopes constitutes antigenic site I of gD. Using a panel of neutralization-resistant mutants, we previously found that group I MAbs can be divided into two subgroups, Ia and Ib, such that mutations selected with Ia antibodies have little or no effect on binding and neutralization by Ib antibodies, and vice versa. Antigenic site I therefore consists of two parts, Ia and Ib. We have now identified the point mutations which prevent neutralization. Two Ib MAbs (DL11 and 4S) selected a Ser to Asn change at residue 140; this alteration creates a new N-linked glycosylation site, which is used. A third Ib MAb (D2) selected a Gln to Leu change at 132. The mutation selected by the Ia MAb HD1 (Ser to Asn at residue 216) is identical to that selected by MAb LP2, another Ia antibody. By using oligonucleotide-directed mutagenesis, we have produced gD genes with combinations of the above mutations. Attempts to recombine these genes into the virus genome were unsuccessful, suggesting that the combinations are lethal. This was confirmed by a complementation assay which measures the ability of gD transiently expressed in transfected Vero cells to rescue the production of infectious virus by the gD-minus mutant F-gD beta.
Muggeridge
M I
MI
Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104.
Wu
T T
TT
Johnson
D C
DC
Glorioso
J C
JC
Eisenberg
R J
RJ
Cohen
G H
GH
D001483:Base Sequence; D003001:Cloning, Molecular; D005814:Genes, Viral; D005816:Genetic Complementation Test; D006031:Glycosylation; D008969:Molecular Sequence Data; D009154:Mutation; D009500:Neutralization Tests; D011995:Recombination, Genetic; D018139:Simplexvirus / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000502:physiology
D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1