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The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells

As a result of a viral infection, viral genomes are not only recognized by RIG-I, but also lead to the activation of RNase L, which cleaves cellular RNA to generate the endogenous RIG-I ligand (eRL). The eRL was previously identified as a specific sequence derived from the internal transcribed space...

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Autores principales: Steinberg, Julia, Wadenpohl, Timo, Jung, Stephanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402674/
https://www.ncbi.nlm.nih.gov/pubmed/34452429
http://dx.doi.org/10.3390/v13081564
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author Steinberg, Julia
Wadenpohl, Timo
Jung, Stephanie
author_facet Steinberg, Julia
Wadenpohl, Timo
Jung, Stephanie
author_sort Steinberg, Julia
collection PubMed
description As a result of a viral infection, viral genomes are not only recognized by RIG-I, but also lead to the activation of RNase L, which cleaves cellular RNA to generate the endogenous RIG-I ligand (eRL). The eRL was previously identified as a specific sequence derived from the internal transcribed spacer region 2, which bears a 2′3′ cyclic phosphate instead of the common 5′ triphosphate. By now, the generation of the eRL and its immunostimulatory effect were shown both in vitro and in reporter systems. In this work, we aimed to elucidate whether the eRL is also generated in Influenza A (IAV) and vesicular stomatitis virus (VSV) infected cells. RNA was extracted from virus-infected cells and used for immunostimulations as well as specific PCR-strategies to detect eRL cleavage. We show that the eRL is generated in IAV infected HEK293 cells, but we could not detect specific eRL fragments in VSV infected cells. Further, RIG-I mediated IFN-response depends not only on viral genomes but also on the eRL, as immunostimulatory properties remain present under 5′triphosphate degrading conditions. In summary, we prove the IAV infection induced eRL generation in HEK293 cells, amplifying the innate immune response.
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spelling pubmed-84026742021-08-29 The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells Steinberg, Julia Wadenpohl, Timo Jung, Stephanie Viruses Article As a result of a viral infection, viral genomes are not only recognized by RIG-I, but also lead to the activation of RNase L, which cleaves cellular RNA to generate the endogenous RIG-I ligand (eRL). The eRL was previously identified as a specific sequence derived from the internal transcribed spacer region 2, which bears a 2′3′ cyclic phosphate instead of the common 5′ triphosphate. By now, the generation of the eRL and its immunostimulatory effect were shown both in vitro and in reporter systems. In this work, we aimed to elucidate whether the eRL is also generated in Influenza A (IAV) and vesicular stomatitis virus (VSV) infected cells. RNA was extracted from virus-infected cells and used for immunostimulations as well as specific PCR-strategies to detect eRL cleavage. We show that the eRL is generated in IAV infected HEK293 cells, but we could not detect specific eRL fragments in VSV infected cells. Further, RIG-I mediated IFN-response depends not only on viral genomes but also on the eRL, as immunostimulatory properties remain present under 5′triphosphate degrading conditions. In summary, we prove the IAV infection induced eRL generation in HEK293 cells, amplifying the innate immune response. MDPI 2021-08-07 /pmc/articles/PMC8402674/ /pubmed/34452429 http://dx.doi.org/10.3390/v13081564 Text en © 2021 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
Steinberg, Julia
Wadenpohl, Timo
Jung, Stephanie
The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells
title The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells
title_full The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells
title_fullStr The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells
title_full_unstemmed The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells
title_short The Endogenous RIG-I Ligand Is Generated in Influenza A-Virus Infected Cells
title_sort endogenous rig-i ligand is generated in influenza a-virus infected cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402674/
https://www.ncbi.nlm.nih.gov/pubmed/34452429
http://dx.doi.org/10.3390/v13081564
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