title: High level expression of equine herpesvirus 1 glycoproteins D and H and their role in protection against virus challenge in the C3H (H-2Kk) murine model.
abstract: N and C-terminal truncated forms of equine herpesvirus 1 (EHV 1) glycoproteins gD and gH were expressed in baculovirus resulting in the production of secreted recombinant proteins. A carboxy-terminal histidine tag was included on each of the genes for protein isolation by nickel affinity chromatography. Recombinant gD was recognized by three gD specific monoclonal antibodies, 20C4, 5H6 and F3132. F3132 is a conformationally dependent monoclonal antibody with virus neutralizing activity. Expression of gH was confirmed by reacting the protein with the gH peptide specific antiserum R319. The truncated gD gene was also expressed as a beta-galactosidase fusion protein which was purified from E. coli by nickel affinity chromatography. C3H mice were inoculated with purified recombinant gD or gH or insect cells which had been infected with recombinant baculoviruses. Mice were subsequently challenged with EHV 1. Purified recombinant baculovirus gD provided the most protection and produced high levels of virus neutralizing antibodies. The gD fusion protein was less effective at protecting mice and insect cells infected with either of the recombinant baculoviruses or purified recombinant gH were poor at conferring protection. The results emphasize the importance of using purified proteins in vaccine formulations and of including EHV 1 gD as a component of a subunit vaccine.
authors.affiliation: Molecular Virology Group, Department of Microbiology, University of Leeds, UK. asto@mail.nerc-oxford.ac.uk
authors.forename: A
authors.initials: A
authors.lastname: Stokes
authors.forename: R S
authors.initials: RS
authors.lastname: Cameron
authors.forename: R N
authors.initials: RN
authors.lastname: Marshall
authors.forename: R A
authors.initials: RA
authors.lastname: Killington
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016367:Baculoviridae / Q000235:genetics; D004195:Disease Models, Animal; D005822:Genetic Vectors / Q000378:metabolism; D006566:Herpesviridae Infections / Q000517:prevention & control*; D004861:Herpesvirus 1, Equid / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008168:Lung / Q000821:virology; D051379:Mice; D008809:Mice, Inbred C3H; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000276:immunology; D014420:Turbinates / Q000821:virology; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D015431:Weight Loss / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C488988:glycoprotein D, Equid herpesvirus 1; C108360:glycoprotein H, Equid herpesvirus 1