Equine herpesvirus 1 glycoprotein D expressed in E. coli provides partial protection against equine herpesvirus infection in mice and elicits virus-neutralizing antibodies in the horse.
The envelope glycoprotein D of EHV-1 (EHV-1 gD) is essential for virus infectivity and entry of virus into cells and is a potent inducer of virus-neutralizing antibody. In this study, truncated EHV-1 gD (gDt) was expressed with a C-terminal hexahistidine tag in E. coli using a pET vector. Western blot analysis using an anti-gD monoclonal antibody demonstrated the presence of gDt bands at 37.5, 36, 29.5 and 28 kDa. The immunogenicity and protective efficacy of partially purified gDt was compared with gD expressed in insect cells by a recombinant baculovirus (Bac gD) using a BALB/c mouse model of EHV-1 respiratory infection. The proteins were also compared in a prime-boost protocol following an initial inoculation with gD DNA. gDt elicited similar levels of gD-specific antibody and neutralizing antibody compared with Bac gD and also provided a similar level of protection against EHV-1 challenge in mice. Inoculation of horses with gDt elicited EHV-1 gD-specific antibodies including virus-neutralizing antibody, suggesting that despite the lack of glycosylation, E. coli may be a useful vehicle for large scale production of EHV-1 gD for vaccine studies.
Weerasinghe
C U
CU
Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, Australia.
Learmonth
G S
GS
Gilkerson
J R
JR
Foote
C E
CE
Wellington
J E
JE
Whalley
J M
JM
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western / Q000662:veterinary; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006566:Herpesviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary* / Q000821:virology; D004861:Herpesvirus 1, Equid / Q000235:genetics / Q000276:immunology*; D006734:Horse Diseases / Q000276:immunology / Q000517:prevention & control* / Q000821:virology*; D006736:Horses; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D012140:Respiratory Tract Diseases / Q000276:immunology / Q000517:prevention & control* / Q000662:veterinary* / Q000821:virology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*
D000914:Antibodies, Viral; D004279:DNA, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C488988:glycoprotein D, Equid herpesvirus 1