title: Immunoselection of recombinant baculoviruses expressing high levels of biologically active herpes simplex virus type 1 glycoprotein D.
abstract: The DNA sequence encoding the complete herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) was inserted into a baculovirus transfer vector under control of the polyhedrin gene promoter of the baculovirus Autographa california nuclear polyhedrosis virus (AcNPV). After co-transfection of Spodoptera frugiperda (Sf9) insect cells with wild-type AcNPV DNA and the recombinant transfer vector DNA, polyhedrin-negative recombinants that expressed high levels of HSV-1 gD were isolated using immunoaffinity selection with antibody coated magnetic particles followed by plaque purification. These recombinant baculoviruses expressed a protein that was slightly smaller than virion HSV-1 gD made in Vero cells. This recombinant protein was expressed at high levels. The expressed protein was glycosylated, was found on the membrane of Sf9 cells, and reacted with gD specific antibodies. Antibodies raised in mice to the recombinant gD neutralized HSV-1 as measured by plaque reduction assays. Mice inoculated with the recombinant baculovirus were completely protected from lethal challenge with HSV-1.
authors.affiliation: Ophthalmology Research, Cedars-Sinai Medical Center, Los Angeles, California.
authors.forename: H
authors.initials: H
authors.lastname: Ghiasi
authors.forename: A B
authors.initials: AB
authors.lastname: Nesburn
authors.forename: R
authors.initials: R
authors.lastname: Kaiwar
authors.forename: S L
authors.initials: SL
authors.lastname: Wechsler
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006031:Glycosylation; D006561:Herpes Simplex / Q000517:prevention & control; D022283:Herpesvirus Vaccines; D007116:Immunization, Passive; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009036:Moths; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018139:Simplexvirus / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014765:Viral Vaccines / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D004279:DNA, Viral; D022283:Herpesvirus Vaccines; D011994:Recombinant Proteins; D014765:Viral Vaccines; C069093:herpes simplex virus-1 glycoprotein D vaccine