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...
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MDPI AG
2021-12-01
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Online Access: | https://www.mdpi.com/1999-4915/13/12/2433 |
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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. |
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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 |
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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 |
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