Expression and characterization of baculovirus expressed herpes simplex virus type 1 glycoprotein L.
We have constructed a recombinant baculovirus expressing high levels of the herpes simplex virus type 1 (HSV-1) glycoprotein L (gL) in Sf9 cells. Sf9 cells infected with this recombinant virus synthesized three polypeptides of 26-27 kDa 28 kDa, and 31 kDa. The 28 and 31 kDa species were sensitive to tunicamycin and N-glycosidase F (PNGase F) treatment, suggesting that they were glycosylated. As shown by both indirect immunofluorescence and Western blot analysis, using polyclonal antibodies to synthetic gL peptides indicated that the baculovirus expressed gL was abundant on the surface of baculovirus gL infected Sf9 cells. A small fraction of the 31 kDa polypeptide was secreted into the extracellular medium as judged by Western blot analysis. The secreted form of gL was completely resistant to Endoglycosidase H (Endo-H), while the membrane associated form of gL was only partially resistant to Endo-H treatment, suggesting that the secreted gL represented a subpopulation of the membrane bound gL. Mice vaccinated with baculovirus expressed gL produced serum antibodies that reacted with authentic HSV-1 gL. However, these mice produced no HSV-1 neutralizing antibody (titer < 1:10) and they were not protected from lethal intraperitoneal or lethal ocular challenge with HSV-1. Thus, when used as a vaccine in the mouse model, gL, similar to our findings with HSV-1 gH, but unlike our results with the other 6 HSV-1 glycoproteins that we have expressed in this baculovirus system, did not provide any protection against HSV-1 challenge.
Ghiasi
H
H
Cedars-Sinai Research Institute, Los Angeles, California.
Kaiwar
R
R
Slanina
S
S
Nesburn
A B
AB
Wechsler
S L
SL
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D005822:Genetic Vectors; D006031:Glycosylation; D006561:Herpes Simplex / Q000517:prevention & control / Q000821:virology; D016849:Keratitis, Herpetic / Q000517:prevention & control / Q000821:virology; D051379:Mice; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000096:biosynthesis*; D018139:Simplexvirus / Q000276:immunology / Q000502:physiology; D018411:Spodoptera; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology*; D014765:Viral Vaccines; D017735:Virus Latency
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C077407:glycoprotein L, Human herpesvirus 1; D014415:Tunicamycin