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D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses

Influenza A viruses (IAV) modulate host antiviral responses to promote viral growth and pathogenicity. The non-structural (NS1) protein of influenza A virus has played an indispensable role in the inhibition of host immune responses, especially in limiting interferon (IFN) production. In this study,...

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Autores principales: Yu, Mengqi, Guo, Yanna, Zhao, Lingcai, Lu, Yuanlu, Liu, Qingzheng, Li, Yinjing, Deng, Lulu, Shi, Zhiyu, Wang, Haifeng, Dankar, Samar, Ping, Jihui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9228823/
https://www.ncbi.nlm.nih.gov/pubmed/35746676
http://dx.doi.org/10.3390/v14061206
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author Yu, Mengqi
Guo, Yanna
Zhao, Lingcai
Lu, Yuanlu
Liu, Qingzheng
Li, Yinjing
Deng, Lulu
Shi, Zhiyu
Wang, Haifeng
Dankar, Samar
Ping, Jihui
author_facet Yu, Mengqi
Guo, Yanna
Zhao, Lingcai
Lu, Yuanlu
Liu, Qingzheng
Li, Yinjing
Deng, Lulu
Shi, Zhiyu
Wang, Haifeng
Dankar, Samar
Ping, Jihui
author_sort Yu, Mengqi
collection PubMed
description Influenza A viruses (IAV) modulate host antiviral responses to promote viral growth and pathogenicity. The non-structural (NS1) protein of influenza A virus has played an indispensable role in the inhibition of host immune responses, especially in limiting interferon (IFN) production. In this study, random site mutations were introduced into the NS1 gene of A/WSN/1933 (WSN, H1N1) via an error prone PCR to construct a random mutant plasmid library. The NS1 random mutant virus library was generated by reverse genetics. To screen out the unidentified NS1 functional mutants, the library viruses were lung-to-lung passaged in mice and individual plaques were picked from the fourth passage in mice lungs. Sanger sequencing revealed that eight different kinds of mutations in the NS1 gene were obtained from the passaged library virus. We found that the NS1 F9Y mutation significantly enhanced viral growth in vitro (MDCK and A549 cells) and in vivo (BALB/c mice) as well as increased virulence in mice. The NS1 D2I mutation attenuated the viral replication and pathogenicity in both in vitro and in vivo models. Further studies demonstrated that the NS1 F9Y mutant virus exhibited systematic and selective inhibition of cytokine responses as well as inhibited the expression of IFN. In addition, the expression levels of innate immunity-related cytokines were significantly up-regulated after the rNS1 D2I virus infected A549 cells. Collectively, our results revealed that the two mutations in the N-terminal of the NS1 protein could alter the viral properties of IAV and provide additional evidence that the NS1 protein is a critical virulence factor. The two characterized NS1 mutations may serve as potential targets for antiviral drugs as well as attenuated vaccine development.
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spelling pubmed-92288232022-06-25 D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses Yu, Mengqi Guo, Yanna Zhao, Lingcai Lu, Yuanlu Liu, Qingzheng Li, Yinjing Deng, Lulu Shi, Zhiyu Wang, Haifeng Dankar, Samar Ping, Jihui Viruses Article Influenza A viruses (IAV) modulate host antiviral responses to promote viral growth and pathogenicity. The non-structural (NS1) protein of influenza A virus has played an indispensable role in the inhibition of host immune responses, especially in limiting interferon (IFN) production. In this study, random site mutations were introduced into the NS1 gene of A/WSN/1933 (WSN, H1N1) via an error prone PCR to construct a random mutant plasmid library. The NS1 random mutant virus library was generated by reverse genetics. To screen out the unidentified NS1 functional mutants, the library viruses were lung-to-lung passaged in mice and individual plaques were picked from the fourth passage in mice lungs. Sanger sequencing revealed that eight different kinds of mutations in the NS1 gene were obtained from the passaged library virus. We found that the NS1 F9Y mutation significantly enhanced viral growth in vitro (MDCK and A549 cells) and in vivo (BALB/c mice) as well as increased virulence in mice. The NS1 D2I mutation attenuated the viral replication and pathogenicity in both in vitro and in vivo models. Further studies demonstrated that the NS1 F9Y mutant virus exhibited systematic and selective inhibition of cytokine responses as well as inhibited the expression of IFN. In addition, the expression levels of innate immunity-related cytokines were significantly up-regulated after the rNS1 D2I virus infected A549 cells. Collectively, our results revealed that the two mutations in the N-terminal of the NS1 protein could alter the viral properties of IAV and provide additional evidence that the NS1 protein is a critical virulence factor. The two characterized NS1 mutations may serve as potential targets for antiviral drugs as well as attenuated vaccine development. MDPI 2022-06-01 /pmc/articles/PMC9228823/ /pubmed/35746676 http://dx.doi.org/10.3390/v14061206 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yu, Mengqi
Guo, Yanna
Zhao, Lingcai
Lu, Yuanlu
Liu, Qingzheng
Li, Yinjing
Deng, Lulu
Shi, Zhiyu
Wang, Haifeng
Dankar, Samar
Ping, Jihui
D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses
title D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses
title_full D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses
title_fullStr D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses
title_full_unstemmed D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses
title_short D2I and F9Y Mutations in the NS1 Protein of Influenza A Virus Affect Viral Replication via Regulating Host Innate Immune Responses
title_sort d2i and f9y mutations in the ns1 protein of influenza a virus affect viral replication via regulating host innate immune responses
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9228823/
https://www.ncbi.nlm.nih.gov/pubmed/35746676
http://dx.doi.org/10.3390/v14061206
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