EHV-1과 EHV-4의 subunit vaccine과 혈청학적 진단법에 대한 연구

Abstract

Equine herpesvirus (EHV) is one of the major cause of economic loss of the equine industry. There is a continuing need to develop efficient diagnostic tool and vaccine to diagnose and prevent EHV infection. After conditioning of high titer production of EHV, we could obtain the viruses up to 1011.75 TCID50/ml. We have constructed 22 hybridoma clones secreting monoclonal antibodies (MAbs) specific to EHV by fusion of mouse myeloma cells with lymphocytes of mice immunized with EHV-1 and EHV-4. The envelope glycoprotein D (gD) of EHV-1, 4 are essential for virus infectivity and are a potent inducer of virus-neutralizing antibody. To develop an efficient subunit vaccine, gD of EHV-1, 4 were expressed with C-terminal hexa-histidine tag in E. coli using pQE30 and pQEgfpⅡ vectors. SDS-PAGE analysis using expressed EHV-1 gD demonstrated the presence of gD bands at 80 kDa, 50 kDa and SDS-PAGE analysis using expressed EHV-4 gD demonstrated the presence of gD bands at 75 kDa, 45 kDa. Two gDs of EHV-1 and -4 were ninety nine percent homologous based on amino acid sequence. MAbs and recombinant gDs developed in this study could be applied as the source materials for diagnostic tools and subunit vaccines to control the EHV epidemic in equine industry, and additional research need to be continued for industrialization.