Interferon-Induced Protection against Foot-and-Mouth Disease Virus Infection Correlates with Enhanced Tissue-Specific Innate Immune Cell Infiltration and Interferon-Stimulated Gene Expression

ABSTRACT Previously, we demonstrated that type I interferon (IFN-α/β) or a combination of IFN-α/β and type II IFN (IFN-γ) delivered by a replication-defective human adenovirus 5 (Ad5) vector protected swine when challenged 1 day later with foot-and-mouth disease virus (FMDV). To gain a more comprehensive understanding of the mechanism of protection induced by IFNs, we inoculated groups of six swine with Ad5-vectors containing these genes, challenged 1 day later and euthanized 2 animals from each group prior to (1 day postinoculation [dpi]) and at 1 (2 dpi) and 6 days postchallenge (7 dpi). Blood, skin, and lymphoid tissues were examined for IFN-stimulated gene (ISG) induction and infiltration by innate immune cells. All IFN-inoculated animals had delayed and decreased clinical signs and viremia compared to the controls, and one animal in the IFN-α treated group did not develop disease. At 1 and 2 dpi the groups inoculated with the IFNs had increased numbers of dendritic cells and natural killer cells in the skin and lymph nodes, respectively, as well as increased levels of several ISGs compared to the controls. In particular, all tissues examined from IFN-treated groups had significant upregulation of the chemokine 10-kDa IFN-γ-inducible protein 10, and preferential upregulation of 2′,5′-oligoadenylate synthetase, Mx1, and indoleamine 2,3-dioxygenase. There was also upregulation of monocyte chemotactic protein 1 and macrophage inflammatory protein 3α in the skin. These data suggest that there is a complex interplay between IFN-induced immunomodulatory and antiviral activities in protection of swine against FMDV.

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Impairment of the DeISGylation Activity of Foot-and-Mouth Disease Virus Lpro Causes Attenuation In Vitro and In Vivo

Journal of Virology ◽

10.1128/jvi.00341-20 ◽

2020 ◽

Vol 94 (13) ◽

Cited By ~ 4

Author(s):

Gisselle N. Medina ◽

Paul Azzinaro ◽

Elizabeth Ramirez-Medina ◽

Joseph Gutkoska ◽

Ying Fang ◽

...

Keyword(s):

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Type I ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Infected Cells ◽

Viral Attenuation

ABSTRACT Foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) affects several pathways of the host innate immune response. Previous studies in bovine cells demonstrated that deletions (leaderless [LLV]) or point mutations in Lpro result in increased expression of interferon (IFN) and IFN-stimulated genes (ISGs), including, among others, the ubiquitin-like protein modifier ISG15 and the ubiquitin specific peptidase USP18. In addition to its conventional papain-like protease activity, Lpro acts as a deubiquitinase (DUB) and deISGylase. In this study, we identified a conserved residue in Lpro that is involved in its interaction with ISG15. Mutation W105A rendered Escherichia coli-expressed Lpro unable to cleave the synthetic substrate pro-ISG15 while preserving cellular eIF4G cleavage. Interestingly, mutant FMDV W105A was viable. Overexpression of ISG15 and the ISGylation machinery in porcine cells resulted in moderate inhibition of FMDV replication, along with a decrease of the overall state of ISGylation in wild-type (WT)-infected cells. In contrast, reduced deISGylation was observed upon infection with W105A and leaderless virus. Reduction in the levels of deubiquitination was also observed in cells infected with the FMDV LproW105A mutant. Surprisingly, similarly to WT, infection with W105A inhibited IFN/ISG expression despite displaying an attenuated phenotype in vivo in mice. Altogether, our studies indicate that abolishing/reducing the deISGylase/DUB activity of Lpro causes viral attenuation independently of its ability to block the expression of IFN and ISG mRNA. Furthermore, our studies highlight the potential of ISG15 to be developed as a novel biotherapeutic molecule against FMD. IMPORTANCE In this study, we identified an aromatic hydrophobic residue in foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) (W105) that is involved in the interaction with ISG15. Mutation in Lpro W105 (A12-LproW105A) resulted in reduced deISGylation in vitro and in porcine-infected cells. Impaired deISGylase activity correlated with viral attenuation in vitro and in vivo and did not affect the ability of Lpro to block expression of type I interferon (IFN) and other IFN-stimulated genes. Moreover, overexpression of ISG15 resulted in the reduction of FMDV viral titers. Thus, our study highlights the potential use of Lpro mutants with modified deISGylase activity for development of live attenuated vaccine candidates, and ISG15 as a novel biotherapeutic against FMD.

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Innate Immune Defenses Induced by CpG Do Not Promote Vaccine-Induced Protection against Foot-and-Mouth Disease Virus in Pigs

Clinical and Vaccine Immunology ◽

10.1128/cvi.00018-09 ◽

2009 ◽

Vol 16 (8) ◽

pp. 1151-1157 ◽

Cited By ~ 25

Author(s):

M. P. Alves ◽

L. Guzylack-Piriou ◽

V. Juillard ◽

J.-C. Audonnet ◽

T. Doel ◽

...

Keyword(s):

Disease Virus ◽

Large Scale ◽

Control Strategies ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Innate Immune ◽

Noticeable Effect ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Natural Hosts

ABSTRACT Emergency vaccination as part of the control strategies against foot-and-mouth disease virus (FMDV) has the potential to limit virus spread and reduce large-scale culling. To reduce the time between vaccination and the onset of immunity, immunostimulatory CpG was tested for its capacity to promote early protection against FMDV challenge in pigs. To this end, CpG 2142, an efficient inducer of alpha interferon, was injected intramuscularly. Increased transcription of Mx1, OAS, and IRF-7 was identified as a sensitive measurement of CpG-induced innate immunity, with increased levels detectable to at least 4 days after injection of CpG formulated with Emulsigen. Despite this, CpG combined with an FMD vaccine did not promote protection. Pigs vaccinated 2 days before challenge had disease development, which was at least as acute as that of unvaccinated controls. All pigs vaccinated 7 days before challenge were protected without a noticeable effect of CpG. In summary, our results demonstrate the caution required when translating findings from mouse models to natural hosts of FMDV.

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Synthetic RNAs Mimicking Structural Domains in the Foot-and-Mouth Disease Virus Genome Elicit a Broad Innate Immune Response in Porcine Cells Triggered by RIG-I and TLR Activation

Viruses ◽

10.3390/v7072807 ◽

2015 ◽

Vol 7 (7) ◽

pp. 3954-3973 ◽

Cited By ~ 14

Author(s):

Belén Borrego ◽

Miguel Rodríguez-Pulido ◽

Concepción Revilla ◽

Belén Álvarez ◽

Francisco Sobrino ◽

...

Keyword(s):

Immune Response ◽

Disease Virus ◽

Innate Immune Response ◽

Foot And Mouth Disease ◽

Virus Genome ◽

Mouth Disease ◽

Innate Immune ◽

Structural Domains ◽

Mouth Disease Virus ◽

Foot And Mouth

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Porcine Type I Interferon Rapidly Protects Swine Against Challenge with Multiple Serotypes of Foot-and-Mouth Disease Virus

Journal of Interferon & Cytokine Research ◽

10.1089/jir.2010.0055 ◽

2011 ◽

Vol 31 (2) ◽

pp. 227-236 ◽

Cited By ~ 43

Author(s):

Camila C.A. Dias ◽

Mauro P. Moraes ◽

Fayna Diaz-San Segundo ◽

Teresa de los Santos ◽

Marvin J. Grubman

Keyword(s):

Disease Virus ◽

Type I Interferon ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Type I ◽

Mouth Disease Virus ◽

Foot And Mouth

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Differential Persistence of Foot-and-Mouth Disease Virus in African Buffalo Is Related to Virus Virulence

Journal of Virology ◽

10.1128/jvi.00166-16 ◽

2016 ◽

Vol 90 (10) ◽

pp. 5132-5140 ◽

Cited By ~ 29

Author(s):

Francois Maree ◽

Lin-Mari de Klerk-Lorist ◽

Simon Gubbins ◽

Fuquan Zhang ◽

Julian Seago ◽

...

Keyword(s):

Disease Virus ◽

Foot And Mouth Disease ◽

Germinal Centers ◽

Cell Killing ◽

Mouth Disease ◽

Lymphoid Tissues ◽

African Buffalo ◽

Mouth Disease Virus ◽

Foot And Mouth

ABSTRACTFoot-and-mouth disease (FMD) virus (FMDV) circulates as multiple serotypes and strains in many regions of endemicity. In particular, the three Southern African Territories (SAT) serotypes are maintained effectively in their wildlife reservoir, the African buffalo, and individuals may harbor multiple SAT serotypes for extended periods in the pharyngeal region. However, the exact site and mechanism for persistence remain unclear. FMD in buffaloes offers a unique opportunity to study FMDV persistence, as transmission from carrier ruminants has convincingly been demonstrated for only this species. Following coinfection of naive African buffaloes with isolates of three SAT serotypes from field buffaloes, palatine tonsil swabs were the sample of choice for recovering infectious FMDV up to 400 days postinfection (dpi). Postmortem examination identified infectious virus for up to 185 dpi and viral genomes for up to 400 dpi in lymphoid tissues of the head and neck, focused mainly in germinal centers. Interestingly, viral persistencein vivowas not homogenous, and the SAT-1 isolate persisted longer than the SAT-2 and SAT-3 isolates. Coinfection and passage of these SAT isolates in goat and buffalo cell lines demonstrated a direct correlation between persistence and cell-killing capacity. These data suggest that FMDV persistence occurs in the germinal centers of lymphoid tissue but that the duration of persistence is related to virus replication and cell-killing capacity.IMPORTANCEFoot-and-mouth disease virus (FMDV) causes a highly contagious acute vesicular disease in domestic livestock and wildlife species. African buffaloes (Syncerus caffer) are the primary carrier hosts of FMDV in African savannah ecosystems, where the disease is endemic. We have shown that the virus persists for up to 400 days in buffaloes and that there is competition between viruses during mixed infections. There was similar competition in cell culture: viruses that killed cells quickly persisted more efficiently in passaged cell cultures. These results may provide a mechanism for the dominance of particular viruses in an ecosystem.

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Inhibition of MAVS Aggregation-Mediated Type-I Interferon Signaling by Foot-and-Mouth Disease Virus VP3

Viruses ◽

10.3390/v13091776 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1776

Author(s):

Pathum Ekanayaka ◽

Byeong-Hoon Lee ◽

Asela Weerawardhana ◽

Kiramage Chathuranga ◽

Jong-Hyeon Park ◽

...

Keyword(s):

Disease Virus ◽

Virus Assembly ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Clear Understanding ◽

Type I ◽

Type I Ifn ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Tm Domain

As a structural protein of the Foot-and-mouth disease virus (FMDV), VP3 plays a vital role in virus assembly and inhibiting the interferon (IFN) signal transduction to promote FMDV replication. Previous studies demonstrated that FMDV VP3 blocks the type-I IFN response by inhibiting the mRNA expression of the mitochondrial antiviral-signaling protein (MAVS); however, the underlying mechanism is poorly understood. Here, we describe the specificity of FMDV VP3 interaction with the transmembrane (TM) domain of MAVS as FMDV driven type-I IFN inhibitory mechanism for its effective replication. The TM domain of MAVS governs the mitochondria localization of MAVS, and it is a key factor in type-I IFN signaling transduction via MAVS aggregation. Thereby, the interaction of FMDV VP3 with the TM domain of MAVS leads to the inhibition of MAVS mitochondria localization, self-association, and aggregation, resulting in the suppression of type-I IFN response. Collectively, these results provide a clear understanding of a key molecular mechanism used by the FMDV VP3 for the suppression of IFN responses via targeting MAVS.

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