Expression and Characterization of Herpes Simplex Virus Type 1 (HSV-1) Glycoprotein G (gG) by Recombinant Vaccinia Virus: Neutralization of HSV-1 Infectivity with Anti-gG Antibody

Abstract

SUMMARYA recombinant vaccinia virus expressing herpes simplex virus type 1 (HSV-1) glycoprotein G (gG) has been constructed. Cells infected with this recombinant virus (gG-VAC) synthesized glycosylated proteins of 48K, 57K and 61K tool. wt. that were recognized by anti-HSV-1 sera. Rabbits and mice vaccinated with the live recombinant virus produced antibodies that recognized 48K, 57K and 61K mol. wt. proteins in HSV-l-infected cells. The gG polypeptides were present on cytoplasmic and nuclear membranes during infection with both HSV-1 and recombinant vaccinia virus gG-VAC. The 57K and 61K tool. wt. gG polypeptides were present in purified HSV-1 virions and were targets for antibody-mediated complement-dependent virus neutralization.

The Herpes Simplex Virus Type 1 US7 Gene Product Is a 66K Glycoprotein and Is a Target for Complement-dependent Virus Neutralization

“…In the absence of fresh complement no virus neutralization was observed. The extent of neutralization was indistinguishable from that obtained in parallel with antisera raised against a vaccinia virus recombinant expressing HSV-1 gG (Sullivan & Smith, 1987). We had noted that anti-HSV-2 antibody reacted with plaques formed by US7-VAC and therefore were interested to know whether anti-US7-VAC would neutralize HSV-2 infectivity.…”

“…Where indicated tunicamycin was added at 10 gg/ml 30 rain before, and during the labelling period. Lysates were prepared from infected cells and treated with antisera as previously described (Sullivan & Smith, 1987). Precipitated polypeptides were electrophoresed on 10~ polyacrylamide gels.…”

“…BHK cells infected with WT-VAC or US7-VAC at 30 p.f.u./cell were harvested and lysed 16 h post-infection. Lysates were run on 10~ polyacrylamide gels and polypeptides transferred to nitrocellulose membranes as described (Sullivan & Smith, 1987). Membranes were incubated with, antisera followed by 1251-labelled affinity-purified Protein A.…”

“…A further five open reading frames with glycoprotein characteristics were identified from the nucleotide sequence of the unique short (Us) region (McGeoch et al, 1985). Open reading frame US6 codes for gD (Spear, 1976), US8 for gE (Baucke & Spear, 1979) and US4 for gG (Ackermann et al, 1986;Richman et al, 1986;Frame et al, 1986;Sullivan & Smith, 1987) which has multiple forms in HSV-l-infected cells and virions. No product has yet been assigned to US5 but very recently US7 was shown to encode a 65K glycoprotein, designated gI (Longnecker et al, 1987).…”

Analysis of the L1 Gene Product of Human Papillomavirus Type 16 by Expression in a Vaccinia Virus Recombinant

“…Vaccinia virus strain WR and the recombinant vaccinia viruses Ll-Vacc and vL1RK were grown in BHK cells and titrated on CV-1 ceils (Mackett et al, 1985). Recombinant gG-VAC was constructed by Sullivan & Smith (1987).…”