Abstract
Foot-and-mouth disease, caused by foot-and-mouth disease virus (FMDV), is an economically devastating disease affecting several important livestock species. FMDV is antigenically diverse and exists as seven serotypes comprised of many strains which are poorly cross-neutralised by antibodies induced by infection or vaccination. Co-infection and recombination are important drivers of antigenic diversity, especially in regions where several serotypes co-circulate at high prevalence, and therefore experimental systems to study these events in vitro would be beneficial. Here we have utilised recombinant FMDVs containing an HA or a FLAG epitope tag within the VP1 capsid protein to investigate the products of co-infection in vitro. Co-infection with viruses from the same and from different serotypes was demonstrated by immunofluorescence microscopy and flow cytometry using anti-tag antibodies. FLAG-tagged VP1 and HA-tagged VP1 could be co-immunoprecipitated from co-infected cells, suggesting that newly synthesised capsids may contain VP1 proteins from both co-infecting viruses. Furthermore, we provide the first demonstration of trans-encapsidation of an FMDV genome into capsids comprised of proteins encoded by a co-infecting heterologous virus. This system provides a useful tool for investigating co-infection dynamics in vitro, particularly between closely related strains, and has the advantage that it does not depend upon the availability of strain-specific FMDV antibodies.
Highlights
We have previously described the construction of epitope-tagged foot-and-mouth disease virus (FMDV) containing either an HA or a FLAG tag located within the GH loop of VP1 [22]
To investigate the feasibility of using these viruses to study co-infection dynamics and trans-encapsidation events in FMDV, we first ascertained whether cells could be simultaneously co-infected with both FLAG- and HA-tagged viruses
Cultures of goat epithelium cells, which express the FMDV integrin αvβ6, were infected with tagged viruses individually or in combination, and tagged VP1 proteins were detected by Western blotting of whole cell lysates using anti-HA and anti-FLAG antibodies (Figure 1b)
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Foot-and-mouth disease virus (FMDV) is the etiologic agent of foot-and-mouth disease (FMD), an economically important vesicular disease affecting even-toed ungulates including critical livestock species such as cattle, sheep and pigs. FMDV is a member of the Aphthovirus genus within the Picornaviridae family of viruses and is comprised of a positive-sense, single-stranded RNA genome of around 8.5 kb inside a small, nonenveloped icosahedral capsid. The genome contains a single open reading frame encoding a polyprotein that is post-translationally processed by viral proteases into the mature viral structural and non-structural proteins [1]. The FMDV capsid contains 60 copies of each of the structural proteins VP1 (1D), VP2
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