Key result
Sera from experimentally or persistently EAV-infected horses immunoprecipitated nsp2, nsp4, nsp5, and nsp12, whereas sera from vaccinated horses reacted consistently only with nsp2 and nsp12.
Different EAV infection states (experimental, persistent, vaccinated) elicit distinct humoral immune responses to viral nonstructural proteins, which may aid in developing improved serologic diagnostics.
Distinct NSP responses by infection status in horses; leaves open DIVA diagnostic utility pending validation studies.
Equine arteritis virus (EAV) replicase consists of two polyproteins (pp1a and pp1ab) that are encoded by open reading frames (ORFs) 1a and 1b of the viral genome. These two replicase polyproteins are posttranslationally processed by three ORF 1a-encoded proteinases to yield at least 13 nonstructural proteins (nsp1 to nsp12, including nsp7α and 7β). These nsps are expressed in EAV-infected cells, but the equine immune response they induce has not been studied. Therefore, the primary purpose of this study was to evaluate the humoral immune response of horses to each of the nsps following EAV infection. Individual nsp coding regions were cloned and expressed in both mammalian and bacterial expression systems. Each recombinant protein was used in an immunoprecipitation assay with equine serum samples from horses (n = 3) that were experimentally infected with three different EAV strains (VB, KY77, and KY84), from stallions (n = 4) that were persistently infected with EAV, and from horses (n = 4) that were vaccinated with the modified live-virus (MLV) vaccine strain. Subsequently, protein-antibody complexes were subjected to Western immunoblotting analysis with individual nsp-specific rabbit antisera, mouse anti-His antibody, or anti-FLAG tag antibody. Nsp2, nsp4, nsp5, and nsp12 were immunoprecipitated by most of the sera from experimentally or persistently infected horses, while sera from vaccinated horses did not react with nsp5 and reacted weakly with nsp4. However, serum samples from vaccinated horses were able to immunoprecipitate nsp2 and nsp12 proteins consistently. Information from this study will assist ongoing efforts to develop improved methods for the serologic diagnosis of EAV infection in horses.
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Go et al. (2010) studied Equine Arteritis Virus (EAV) infection (n=11). EAV infection or vaccination was evaluated on Immunoprecipitation of nonstructural proteins by equine serum samples. Sera from experimentally or persistently EAV-infected horses immunoprecipitated nsp2, nsp4, nsp5, and nsp12, whereas sera from vaccinated horses reacted consistently only with nsp2 and nsp12.
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