title: The production of a truncated form of baculovirus expressed EHV-1 glycoprotein C and its role in protection of C3H (H-2Kk) mice against virus challenge.
abstract: A truncated form of the equine herpesvirus 1 (EHV-1) glycoprotein C (gC) gene was expressed in baculovirus. The gC signal sequence was substituted with the honeybee melittin signal sequence and the transmembrane region was replaced with a histidine tag. The recombinant virus produced high levels of gC in both the cells and supernatants of infected cells. The protein was present by 24 h and maximal secretion occurred at 96 h post-infection. The recombinant protein was antigenically authentic as shown by its reaction with each of a panel of individual monoclonal antibodies specific for the five distinct antigenic sites on EHV-1 gC. Recombinant gC was purified from the supernatant of infected cells by immuno-affinity chromatography and used to immunize C3H (H-2Kk haplotype) mice. This incurred a gC specific antibody response against both the recombinant protein and EHV-1 gC. 'Pepscan' analysis showed that the gC specific antibodies in serum from these mice reacted with the same epitopes on gC as those recognized by antibodies in convalescent equine sera (i.e. antibodies were specific to antigenic sites one and five). A third previously unrecognized antibody binding site at the carboxyl terminus was also detected (Antibody binding domain I). A T-cell proliferative response against EHV-1 was detected in splenocyte populations taken from vaccinated mice. Further, the recovery of virus from the lungs and turbinates following challenge of mice with EHV-1 was significantly reduced. These findings indicate that baculovirus expressed gC may contribute significantly to a subunit vaccine preparation aimed at protecting horses from EHV-1 infection.
authors.affiliation: Department of Microbiology, University of Leeds, UK.
authors.forename: A
authors.initials: A
authors.lastname: Stokes
authors.forename: D G
authors.initials: DG
authors.lastname: Alber
authors.forename: R S
authors.initials: RS
authors.lastname: Cameron
authors.forename: R N
authors.initials: RN
authors.lastname: Marshall
authors.forename: G P
authors.initials: GP
authors.lastname: Allen
authors.forename: R A
authors.initials: RA
authors.lastname: Killington
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D005822:Genetic Vectors; D006566:Herpesviridae Infections / Q000517:prevention & control*; D004861:Herpesvirus 1, Equid / Q000276:immunology*; D051379:Mice; D008809:Mice, Inbred C3H; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D018411:Spodoptera / Q000166:cytology; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C096213:glycoprotein 13, Equid herpesvirus