TY - JOUR
T1 - Antigenic and functional analysis of a neutralization site of HSV-1 glycoprotein D
AU - Muggeridge, Martin I.
AU - Wu, Tsung Teh
AU - Johnson, David C.
AU - Glorioso, Joseph C.
AU - Eisenberg, Roselyn J.
AU - Cohen, Gary H.
N1 - Funding Information:
This investigation was supported by Public Health Service Grants DE-02623 from the National Institute of Dental Research, Al-l 8289 from the National Institute of Allergy and Infectious Diseases, and by a grant to G.H.C. and R.J.E. from the American Cyanamid Co. D.C.J. is a Research Scholar of the National Cancer Institute of Canada. We thank Lenore Pereira and Martin Zweig for monoclonal antibodies, Anthony Minson for antibodies and the MarD.LP2 and MarD.LP14 viruses, Randal Byrn for pMar62. and Lewis Pizer for pJB3.
PY - 1990/2
Y1 - 1990/2
N2 - 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, la and Ib, such that mutations selected with la antibodies have little or no effect on binding and neutralization by Ib antibodies, and vice versa. Antigenic site I therefore consists of two parts, la 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 Gin 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 la 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-gDβ.
AB - 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, la and Ib, such that mutations selected with la antibodies have little or no effect on binding and neutralization by Ib antibodies, and vice versa. Antigenic site I therefore consists of two parts, la 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 Gin 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 la 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-gDβ.
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U2 - 10.1016/0042-6822(90)90091-5
DO - 10.1016/0042-6822(90)90091-5
M3 - Article
C2 - 2154881
AN - SCOPUS:0025022498
SN - 0042-6822
VL - 174
SP - 375
EP - 387
JO - Virology
JF - Virology
IS - 2
ER -