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Inactivating Three Interferon Antagonists Attenuates Pathogenesis of an Enteric Coronavirus.
J Virol. 2020 Aug 17; 94(17)JV

Abstract

Coronaviruses (CoVs) have repeatedly emerged from wildlife hosts and infected humans and livestock animals to cause epidemics with significant morbidity and mortality. CoVs infect various organs, including respiratory and enteric systems, as exemplified by newly emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The constellation of viral factors that contribute to developing enteric disease remains elusive. Here, we investigated CoV interferon antagonists for their contribution to enteric pathogenesis. Using an infectious clone of an enteric CoV, porcine epidemic diarrhea virus (icPEDV), we generated viruses with inactive versions of interferon antagonist nonstructural protein 1 (nsp1), nsp15, and nsp16 individually or combined into one virus designated icPEDV-mut4. Interferon-responsive PK1 cells were infected with these viruses and produced higher levels of interferon responses than were seen with wild-type icPEDV infection. icPEDV-mut4 elicited robust interferon responses and was severely impaired for replication in PK1 cells. To evaluate viral pathogenesis, piglets were infected with either icPEDV or icPEDV-mut4. While the icPEDV-infected piglets exhibited clinical disease, the icPEDV-mut4-infected piglets showed no clinical symptoms and exhibited normal intestinal pathology at day 2 postinfection. icPEDV-mut4 replicated in the intestinal tract, as revealed by detection of viral RNA in fecal swabs, with sequence analysis documenting genetic stability of the input strain. Importantly, icPEDV-mut4 infection elicited IgG and neutralizing antibody responses to PEDV. These results identify nsp1, nsp15, and nsp16 as virulence factors that contribute to the development of PEDV-induced diarrhea in swine. Inactivation of these CoV interferon antagonists is a rational approach for generating candidate vaccines to prevent disease and spread of enteric CoVs, including SARS-CoV-2.IMPORTANCE Emerging coronaviruses, including SARS-CoV-2 and porcine CoVs, can infect enterocytes, cause diarrhea, and be shed in the feces. New approaches are needed to understand enteric pathogenesis and to develop vaccines and therapeutics to prevent the spread of these viruses. Here, we exploited a reverse genetic system for an enteric CoV, porcine epidemic diarrhea virus (PEDV), and outline an approach of genetically inactivating highly conserved viral factors known to limit the host innate immune response to infection. Our report reveals that generating PEDV with inactive versions of three viral interferon antagonists, nonstructural proteins 1, 15, and 16, results in a highly attenuated virus that does not cause diarrhea in animals and elicits a neutralizing antibody response in virus-infected animals. This strategy may be useful for generating live attenuated vaccine candidates that prevent disease and fecal spread of enteric CoVs, including SARS-CoV-2.

Authors+Show Affiliations

Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University Chicago, Maywood, Illinois, USA.Virus and Prion Research Unit, USDA-ARS-National Animal Disease Center, Ames, Iowa, USA alexandra.buckley@usda.gov sbaker1@luc.edu.Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, South Dakota, USA.Virus and Prion Research Unit, USDA-ARS-National Animal Disease Center, Ames, Iowa, USA.Virus and Prion Research Unit, USDA-ARS-National Animal Disease Center, Ames, Iowa, USA.Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University Chicago, Maywood, Illinois, USA alexandra.buckley@usda.gov sbaker1@luc.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

32554697

Citation

Deng, Xufang, et al. "Inactivating Three Interferon Antagonists Attenuates Pathogenesis of an Enteric Coronavirus." Journal of Virology, vol. 94, no. 17, 2020.
Deng X, Buckley AC, Pillatzki A, et al. Inactivating Three Interferon Antagonists Attenuates Pathogenesis of an Enteric Coronavirus. J Virol. 2020;94(17).
Deng, X., Buckley, A. C., Pillatzki, A., Lager, K. M., Faaberg, K. S., & Baker, S. C. (2020). Inactivating Three Interferon Antagonists Attenuates Pathogenesis of an Enteric Coronavirus. Journal of Virology, 94(17). https://doi.org/10.1128/JVI.00565-20
Deng X, et al. Inactivating Three Interferon Antagonists Attenuates Pathogenesis of an Enteric Coronavirus. J Virol. 2020 Aug 17;94(17) PubMed PMID: 32554697.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Inactivating Three Interferon Antagonists Attenuates Pathogenesis of an Enteric Coronavirus. AU - Deng,Xufang, AU - Buckley,Alexandra C, AU - Pillatzki,Angela, AU - Lager,Kelly M, AU - Faaberg,Kay S, AU - Baker,Susan C, Y1 - 2020/08/17/ PY - 2020/3/30/received PY - 2020/6/11/accepted PY - 2020/6/20/pubmed PY - 2020/9/2/medline PY - 2020/6/20/entrez KW - SARS-CoV-2 KW - coronavirus KW - endoribonuclease KW - interferon antagonist KW - nsp1 KW - nsp15 KW - nsp16 KW - pathogenesis KW - porcine epidemic diarrhea virus KW - vaccine JF - Journal of virology JO - J Virol VL - 94 IS - 17 N2 - Coronaviruses (CoVs) have repeatedly emerged from wildlife hosts and infected humans and livestock animals to cause epidemics with significant morbidity and mortality. CoVs infect various organs, including respiratory and enteric systems, as exemplified by newly emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The constellation of viral factors that contribute to developing enteric disease remains elusive. Here, we investigated CoV interferon antagonists for their contribution to enteric pathogenesis. Using an infectious clone of an enteric CoV, porcine epidemic diarrhea virus (icPEDV), we generated viruses with inactive versions of interferon antagonist nonstructural protein 1 (nsp1), nsp15, and nsp16 individually or combined into one virus designated icPEDV-mut4. Interferon-responsive PK1 cells were infected with these viruses and produced higher levels of interferon responses than were seen with wild-type icPEDV infection. icPEDV-mut4 elicited robust interferon responses and was severely impaired for replication in PK1 cells. To evaluate viral pathogenesis, piglets were infected with either icPEDV or icPEDV-mut4. While the icPEDV-infected piglets exhibited clinical disease, the icPEDV-mut4-infected piglets showed no clinical symptoms and exhibited normal intestinal pathology at day 2 postinfection. icPEDV-mut4 replicated in the intestinal tract, as revealed by detection of viral RNA in fecal swabs, with sequence analysis documenting genetic stability of the input strain. Importantly, icPEDV-mut4 infection elicited IgG and neutralizing antibody responses to PEDV. These results identify nsp1, nsp15, and nsp16 as virulence factors that contribute to the development of PEDV-induced diarrhea in swine. Inactivation of these CoV interferon antagonists is a rational approach for generating candidate vaccines to prevent disease and spread of enteric CoVs, including SARS-CoV-2.IMPORTANCE Emerging coronaviruses, including SARS-CoV-2 and porcine CoVs, can infect enterocytes, cause diarrhea, and be shed in the feces. New approaches are needed to understand enteric pathogenesis and to develop vaccines and therapeutics to prevent the spread of these viruses. Here, we exploited a reverse genetic system for an enteric CoV, porcine epidemic diarrhea virus (PEDV), and outline an approach of genetically inactivating highly conserved viral factors known to limit the host innate immune response to infection. Our report reveals that generating PEDV with inactive versions of three viral interferon antagonists, nonstructural proteins 1, 15, and 16, results in a highly attenuated virus that does not cause diarrhea in animals and elicits a neutralizing antibody response in virus-infected animals. This strategy may be useful for generating live attenuated vaccine candidates that prevent disease and fecal spread of enteric CoVs, including SARS-CoV-2. SN - 1098-5514 UR - https://www.unboundmedicine.com/medline/citation/32554697/Inactivating_Three_Interferon_Antagonists_Attenuates_Pathogenesis_of_an_Enteric_Coronavirus_ DB - PRIME DP - Unbound Medicine ER -