title: Characterization of baculovirus-expressed herpes simplex virus type 1 glycoprotein K.
abstract: The DNA region encoding the complete herpes simplex virus type 1 (HSV-1) glycoprotein K (gK) was inserted into a baculovirus transfer vector, and recombinant viruses expressing gK were isolated. Four gK-related recombinant baculovirus-expressed peptides of 29, 35, 38, and 40 kDa were detected with polyclonal antibody to gK. The 35-, 38-, and 40-kDa species were susceptible to tunicamycin treatment, suggesting that they were glycosylated. The 38- and 40-kDa species corresponded to partially glycosylated precursor gK (pgK) and mature gK, respectively. The 29-kDa peptide probably represented a cleaved, unglycosylated peptide. The 35-kDa peptide probably represented a cleaved, glycosylated peptide that may be a precursor to pgK. Indirect immunofluorescence with polyclonal antibody to gK peptides indicated that the recombinant baculovirus-expressed gK was abundant on the surface of the insect cells in which it was expressed. Mice vaccinated with the baculovirus-expressed gK produced very low levels (< 1:10) of HSV-1 neutralizing antibody. Nonetheless, these mice were partially protected from lethal challenge with HSV-1 (75% survival). This protection was significant (P = 0.02). Despite some protection against death, gK-vaccinated mice showed no protection against the establishment of latency. Surprisingly, gK-vaccinated mice that were challenged ocularly with a stromal disease-producing strain of HSV-1 had significantly higher levels of ocular disease (herpes stromal keratitis) than did mock-vaccinated mice. In summary, this is the first report to show that vaccination with HSV-1 gK can provide protection against lethal HSV-1 challenge and that vaccination with an HSV-1 glycoprotein can significantly increase the severity of HSV-1-induced ocular disease.
authors.affiliation: Cedars-Sinai Medical Center Research Institute, Los Angeles, California 90048.
authors.forename: H
authors.initials: H
authors.lastname: Ghiasi
authors.forename: S
authors.initials: S
authors.lastname: Slanina
authors.forename: A B
authors.initials: AB
authors.lastname: Nesburn
authors.forename: S L
authors.initials: SL
authors.lastname: Wechsler
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D005123:Eye / Q000382:microbiology; D005455:Fluorescent Antibody Technique; D006031:Glycosylation / Q000187:drug effects; D006561:Herpes Simplex / Q000517:prevention & control*; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009036:Moths / Q000166:cytology; D009500:Neutralization Tests; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D016019:Survival Analysis; D014415:Tunicamycin / Q000494:pharmacology; D014611:Vaccination; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D017735:Virus Latency
chemical_list: D000914:Antibodies, Viral; D011994:Recombinant Proteins; C072448:UL53 protein, Human herpesvirus 1; D014764:Viral Proteins; D014415:Tunicamycin