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. 2013 Oct;75(10):1317-21.
doi: 10.1292/jvms.13-0168. Epub 2013 Jul 7.

Characterization of glycoproteins in equine herpesvirus-1

Hassan Y A H Mahmoud  1 , Kiyohiko Andoh, Shiho Hattori, Yutaka Terada, Keita Noguchi, Hiroshi Shimoda, Ken Maeda
Affiliations

Characterization of glycoproteins in equine herpesvirus-1

Hassan Y A H Mahmoud et al. J Vet Med Sci. 2013 Oct.

Abstract

In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using horse sera, but that of gE, gH, gK, gL, gM and gN was not. Four MAbs recognized gB, four recognized gC and one recognized gp2. Two MAbs against gB cross-reacted with EHV-4. Interestingly, coexpression of gE and gI and gM and gN enhanced their antigenicity. Furthermore, immunoblot analysis of gp2 showed that different molecular masses of gp2 were recognized by the MAb against gp2 and horse sera against EHV-1. In this study, it was demonstrated that at least six glycoproteins were immunogenic to horses, and coexpression of gE and gI and gM and gN was important for enhancement of antigenicity.

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Figures

Fig. 1.
Fig. 1.
Comparison of gp2, gB and gC expressed in plasmid-transfected 293T cells and EHV-1-infected FHK-Tcl3.1 cells. Immunoblot analysis was carried out under non-reducing conditions. MAbs, 8H11(A), 5F12 (B) and 1G10 (C) were used as primary antibodies.
Fig. 2.
Fig. 2.
Co-expression of EHV-1 glycoproteins. Plasmids pCAG-gE-1 and pCAG-gI-1 (A), pCAG-gM-1 and pCAG-gN-1 (B) and pCAG-gH-1 and pCAG-gL-1 (C) were co-transfected into 293T cells. Immunoblot analysis was carried out under non-reducing conditions. Sera collected from horses experimentally infected with EHV-1 were used as primary antibody. The amount of plasmid DNA transfected into 293T cells in 35 mm dishes is noted under each figure.
Fig. 3.
Fig. 3.
(A) Comparison of co-expressed gE and gI by immunoblot analysis using mouse sera specific to gE (169-201) and horse sera specific to EHV-1. Closed arrowheads show gE-specific bands and open arrowheads do gI-specific bands. (B) Comparison of molecular masses of gp2 detected by MAb 8H11 and horse sera specific to EHV-1. Immunoblot analysis was carried out under non-reducing conditions. Closed arrowheads show gp2-specific bands.
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