Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease Viruses

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...

Full description

Bibliographic Details
Main Authors: Kay Childs, Nicholas Juleff, Katy Moffat, Julian Seago
Format: Article
Language:English
Published: MDPI AG 2021-12-01
Series:Viruses
Subjects:
Online Access:https://www.mdpi.com/1999-4915/13/12/2433
_version_ 1797499951066906624
author Kay Childs
Nicholas Juleff
Katy Moffat
Julian Seago
author_facet Kay Childs
Nicholas Juleff
Katy Moffat
Julian Seago
author_sort Kay Childs
collection DOAJ
description 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.
first_indexed 2024-03-10T03:54:50Z
format Article
id doaj.art-19ccbdbc852340cab52189ff815f3f51
institution Directory Open Access Journal
issn 1999-4915
language English
last_indexed 2024-03-10T03:54:50Z
publishDate 2021-12-01
publisher MDPI AG
record_format Article
series Viruses
spelling doaj.art-19ccbdbc852340cab52189ff815f3f512023-11-23T10:57:52ZengMDPI AGViruses1999-49152021-12-011312243310.3390/v13122433Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease VirusesKay Childs0Nicholas Juleff1Katy Moffat2Julian Seago3The Pirbright Institute, Ash Road, Woking GU24 0NF, Surrey, UKThe Pirbright Institute, Ash Road, Woking GU24 0NF, Surrey, UKThe Pirbright Institute, Ash Road, Woking GU24 0NF, Surrey, UKThe Pirbright Institute, Ash Road, Woking GU24 0NF, Surrey, UKFoot-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.https://www.mdpi.com/1999-4915/13/12/2433foot-and-mouth diseaseFMDVco-infectiontrans-encapsidation
spellingShingle Kay Childs
Nicholas Juleff
Katy Moffat
Julian Seago
Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease Viruses
Viruses
foot-and-mouth disease
FMDV
co-infection
trans-encapsidation
title Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease Viruses
title_full Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease Viruses
title_fullStr Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease Viruses
title_full_unstemmed Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease Viruses
title_short Demonstration of Co-Infection and Trans-Encapsidation of Viral RNA In Vitro Using Epitope-Tagged Foot-and-Mouth Disease Viruses
title_sort demonstration of co infection and trans encapsidation of viral rna in vitro using epitope tagged foot and mouth disease viruses
topic foot-and-mouth disease
FMDV
co-infection
trans-encapsidation
url https://www.mdpi.com/1999-4915/13/12/2433
work_keys_str_mv AT kaychilds demonstrationofcoinfectionandtransencapsidationofviralrnainvitrousingepitopetaggedfootandmouthdiseaseviruses
AT nicholasjuleff demonstrationofcoinfectionandtransencapsidationofviralrnainvitrousingepitopetaggedfootandmouthdiseaseviruses
AT katymoffat demonstrationofcoinfectionandtransencapsidationofviralrnainvitrousingepitopetaggedfootandmouthdiseaseviruses
AT julianseago demonstrationofcoinfectionandtransencapsidationofviralrnainvitrousingepitopetaggedfootandmouthdiseaseviruses