Lenore Pereira

Identification and preliminary mapping with monoclonal antibodies of a herpes simplex virus 2 glycoprotein lacking a known type 1 counterpart

The properties of herpes simplex virus 2 (HSV-2)-specific proteins reactive with monoclonal antibody H966 derived from mice immunized with HSV-2 strain G are reported. The reactive proteins contained in infected cell lysates subjected to electrophoresis in denaturing gels and transferred to nitrocellulose sheets form a relatively sharp band characteristic of Mr 124,000 proteins and a diffuse, more slowly migrating band. Antigens reactive with H966 were detected on the surface of viable, unfixed cells. The electrophoretic mobility of the H966-reactive proteins made in the presence of tunicamycin was more rapid than that of the proteins made in the absence of the drug. Direct evidence that the HSV-2-specific antigen was a glycoprotein emerged from purification of [14C]glucosamine-labeled proteins with similar electrophoretic mobilities by immunoabsorption to H966 bound to Sepharose beads. Analyses of the reactivity of HSV-1 X HSV-2 recombinants indicated the gene specifying the glycoprotein maps in the S component of the DNA. The glycoprotein detected by H966 has no known counterpart in HSV-1 and corresponds to the glycoprotein previously designated as gC of HSV-2 and reported to map to the right of gC specified by HSV-1. Inasmuch as an HSV-2 gene colinear with HSV-1 gC has been reported to specify a glycoprotein currently designated as gC of HSV-2 by Para et al. [J. Virol. 45, 1223-1227 (1983)], the glycoprotein identified by H966 should be designated as gG.

Identification, properties, and gene location of a novel glycoprotein specified by herpes simplex virus 1

We report the identification of a novel herpes simplex virus 1 (HSV-1) glycoprotein reactive with type specific monoclonal antibody H1379. The monoclonal antibody reacted with two broad bands with apparent mol wt of 60K to 68K and 44K to 48K formed by infected cell lysates subjected to electrophoresis in denaturing polyacrylamide gels and electrically transferred to a nitrocellulose sheet. Early in infection the H1379 reactive protein was found in the faster migrating band. The rate of accumulation was highest late in infection and only the slower migrating form incorporates significant amounts of glucosamine. The epitopic site recognized by H1379 was not uniformly distributed among strains. Analyses of HSV-1 X HSV-2 recombinants with monoclonal antibodies to HSV-1 and HSV-2 glycoproteins mapping in the S component of the HSV genomes and marker transfer experiments indicated that the gene specifying the H1379 reactive protein maps within BamHI fragment J to the left of gD most probably within the open reading frame designated as US4 (D. J. McGeoch, A Dolan, S. Donald, and F. J. Rixon, 1985, J. Mol. Biol. 181, 1-13). The gene specifying a recently discovered HSV-2 glycoprotein designated as gG-2 (B. Roizman, B. Norrild, C. Chan, and L. Pereira, 1984, Virology 133, 242-247) maps in the corresponding domain of the HSV-2 genome and marker transfer experiments suggest that the H1379 reactive protein and gG-2 are collinear. We have therefore designated the novel HSV-1 glycoprotein as gG-1.

Monoclonal antibodies for rapid diagnosis and typing of genital herpes infections during pregnancy

Fluorescein-conjugated, type-specific monoclonal antibodies to herpes simplex virus (HSV-1 and HSV-2) and group-specific antibody which recognizes both HSV-1 and HSV-2 were used to detect HSV-infected cells in clinical specimens. Specimens were collected from 66 pregnant women with a past history of herpes genitalis or with suspected lesions. Fourteen of 18 samples from which virus was isolated were positive by immunofluorescence test; four of 18 specimens had an insufficient number of cells (less than 50 per smear) for analysis. In one specimen, a positive reaction by immunofluorescence was not confirmed by virus isolation. HSV was typed directly on smears of clinical specimens in 14 instances: three samples were identified as HSV-1, and 11 as HSV-2. Moreover, the immunofluorescence test was used to type HSV isolates in cell cultures in these cases and 40 additional strains. Examination of viral deoxyribonucleic acid fragments obtained by restriction enzyme digestion confirmed the typings; complete agreement was found among the three methods. Immunofluorescence with monoclonal antibodies is a reliable test for rapid diagnosis and simultaneous typing of genital HSV infections, even in asymptomatic women. With adequate specimens, the specificity and sensitivity of this method approach 100%.