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Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), utilizes the host receptor angiotensin-converting enzyme 2 (ACE2) and the auxiliary receptor Neuropilin-1 (NRP1) to enter host cells. NRP1 has another isoform, NRP2, whose function in COVI...

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Autores principales: Ishitoku, Michinori, Mokuda, Sho, Araki, Kei, Watanabe, Hirofumi, Kohno, Hiroki, Sugimoto, Tomohiro, Yoshida, Yusuke, Sakaguchi, Takemasa, Masumoto, Junya, Hirata, Shintaro, Sugiyama, Eiji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10383177/
https://www.ncbi.nlm.nih.gov/pubmed/37515185
http://dx.doi.org/10.3390/v15071498
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author Ishitoku, Michinori
Mokuda, Sho
Araki, Kei
Watanabe, Hirofumi
Kohno, Hiroki
Sugimoto, Tomohiro
Yoshida, Yusuke
Sakaguchi, Takemasa
Masumoto, Junya
Hirata, Shintaro
Sugiyama, Eiji
author_facet Ishitoku, Michinori
Mokuda, Sho
Araki, Kei
Watanabe, Hirofumi
Kohno, Hiroki
Sugimoto, Tomohiro
Yoshida, Yusuke
Sakaguchi, Takemasa
Masumoto, Junya
Hirata, Shintaro
Sugiyama, Eiji
author_sort Ishitoku, Michinori
collection PubMed
description Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), utilizes the host receptor angiotensin-converting enzyme 2 (ACE2) and the auxiliary receptor Neuropilin-1 (NRP1) to enter host cells. NRP1 has another isoform, NRP2, whose function in COVID-19 has seldom been reported. In addition, although patients with severe cases of COVID-19 often exhibit increased levels of proinflammatory cytokines, the relationship between these cytokines and SARS-CoV-2 proliferation remains unknown. The aim of this study is to clarify the roles of proinflammatory cytokines in Neuropilin expressions and in SARS-CoV-2 infection. To identify the expression patterns of NRP under inflamed and noninflamed conditions, next-generation sequencing (RNA-seq), immunohistochemistry, quantitative real-time PCR, and Western blotting were performed using primary cultured fibroblast-like synoviocytes, MH7A (immortalized cell line of human rheumatoid fibroblast-like synoviocytes), immortalized MRC5 (human embryonic lung fibroblast), and synovial tissues. To measure viral proliferative capacity, SARS-CoV-2 infection experiments were also performed. NRP2 was upregulated in inflamed tissues. Cytokine-stimulated human fibroblast cell lines, such as MH7A and immortalized MRC5, revealed that NRP2 expression increased with co-stimulation of tumor necrosis factor α (TNFα) and interleukin-1 beta (IL-1β) and was suppressed with anti-TNFα antibody alone. TNFα and IL-1β promoted SARS-CoV-2 proliferation and Spike protein binding. The viral proliferation coincided with the expression of NRP2, which was modulated through plasmid transfections. Our results revealed that proinflammatory cytokines, including TNFα, contribute to NRP2 upregulation and SARS-CoV-2 proliferation in host human cells.
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spelling pubmed-103831772023-07-30 Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation Ishitoku, Michinori Mokuda, Sho Araki, Kei Watanabe, Hirofumi Kohno, Hiroki Sugimoto, Tomohiro Yoshida, Yusuke Sakaguchi, Takemasa Masumoto, Junya Hirata, Shintaro Sugiyama, Eiji Viruses Article Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), utilizes the host receptor angiotensin-converting enzyme 2 (ACE2) and the auxiliary receptor Neuropilin-1 (NRP1) to enter host cells. NRP1 has another isoform, NRP2, whose function in COVID-19 has seldom been reported. In addition, although patients with severe cases of COVID-19 often exhibit increased levels of proinflammatory cytokines, the relationship between these cytokines and SARS-CoV-2 proliferation remains unknown. The aim of this study is to clarify the roles of proinflammatory cytokines in Neuropilin expressions and in SARS-CoV-2 infection. To identify the expression patterns of NRP under inflamed and noninflamed conditions, next-generation sequencing (RNA-seq), immunohistochemistry, quantitative real-time PCR, and Western blotting were performed using primary cultured fibroblast-like synoviocytes, MH7A (immortalized cell line of human rheumatoid fibroblast-like synoviocytes), immortalized MRC5 (human embryonic lung fibroblast), and synovial tissues. To measure viral proliferative capacity, SARS-CoV-2 infection experiments were also performed. NRP2 was upregulated in inflamed tissues. Cytokine-stimulated human fibroblast cell lines, such as MH7A and immortalized MRC5, revealed that NRP2 expression increased with co-stimulation of tumor necrosis factor α (TNFα) and interleukin-1 beta (IL-1β) and was suppressed with anti-TNFα antibody alone. TNFα and IL-1β promoted SARS-CoV-2 proliferation and Spike protein binding. The viral proliferation coincided with the expression of NRP2, which was modulated through plasmid transfections. Our results revealed that proinflammatory cytokines, including TNFα, contribute to NRP2 upregulation and SARS-CoV-2 proliferation in host human cells. MDPI 2023-07-03 /pmc/articles/PMC10383177/ /pubmed/37515185 http://dx.doi.org/10.3390/v15071498 Text en © 2023 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
Ishitoku, Michinori
Mokuda, Sho
Araki, Kei
Watanabe, Hirofumi
Kohno, Hiroki
Sugimoto, Tomohiro
Yoshida, Yusuke
Sakaguchi, Takemasa
Masumoto, Junya
Hirata, Shintaro
Sugiyama, Eiji
Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation
title Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation
title_full Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation
title_fullStr Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation
title_full_unstemmed Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation
title_short Tumor Necrosis Factor and Interleukin-1β Upregulate NRP2 Expression and Promote SARS-CoV-2 Proliferation
title_sort tumor necrosis factor and interleukin-1β upregulate nrp2 expression and promote sars-cov-2 proliferation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10383177/
https://www.ncbi.nlm.nih.gov/pubmed/37515185
http://dx.doi.org/10.3390/v15071498
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