Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells

Enterovirus 71 (EV71) is typically transmitted by the oral-faecal route and initiates infection upon crossing the intestinal mucosa. Our limited understanding of the mechanisms by which it crosses the intestinal mucosa has hampered the development of effective therapeutic options. Here, using an RNA...

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Main Authors: Sheng-Lin Chen, Yan-Gang Liu, Yong-Tao Zhou, Ping Zhao, Hao Ren, Man Xiao, Yong-Zhe Zhu, Zhong-Tian Qi
Format: Article
Language:English
Published: Taylor & Francis Group 2019-01-01
Series:Emerging Microbes and Infections
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Online Access:https://www.tandfonline.com/doi/10.1080/22221751.2019.1618686
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author Sheng-Lin Chen
Yan-Gang Liu
Yong-Tao Zhou
Ping Zhao
Hao Ren
Man Xiao
Yong-Zhe Zhu
Zhong-Tian Qi
author_facet Sheng-Lin Chen
Yan-Gang Liu
Yong-Tao Zhou
Ping Zhao
Hao Ren
Man Xiao
Yong-Zhe Zhu
Zhong-Tian Qi
author_sort Sheng-Lin Chen
collection DOAJ
description Enterovirus 71 (EV71) is typically transmitted by the oral-faecal route and initiates infection upon crossing the intestinal mucosa. Our limited understanding of the mechanisms by which it crosses the intestinal mucosa has hampered the development of effective therapeutic options. Here, using an RNA interference screen combined with chemical inhibitors or the overexpression of dominant negative proteins, we found that EV71 entry into Caco-2 cells, a polarized human intestinal epithelial cell line, does not involve clathrin- and caveolae-dependent endocytic pathways or macropinocytosis but requires GTP-binding protein dynamin 2 and cytoskeleton remodelling. The use of siRNAs targeting endophilin family members revealed that endophlin-A2 is essential for the uptake of EV71 particles by Caco-2 cells. Subcellular analysis revealed that internalized EV71 virions largely colocalized with endophilin-A2 at cytomembrane ruffles and in the perinuclear area. Combined with viral entry kinetics, these data suggest that EV71 enters Caco-2 cells mainly via an endophilin-A2-mediated endocytic (EME) pathway. Finally, we showed that internalized EV71 virions were transported to endosomal sorting complex required for transport (ESCRT)-related multivesicular bodies (MVBs). These data provide attractive therapeutic targets to block EV71 infection.
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spelling doaj-art-13cd8661a42c4f74a8ccca3b85b810892025-08-20T01:54:18ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512019-01-018177378610.1080/22221751.2019.1618686Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cellsSheng-Lin Chen0Yan-Gang Liu1Yong-Tao Zhou2Ping Zhao3Hao Ren4Man Xiao5Yong-Zhe Zhu6Zhong-Tian Qi7Department of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University Shanghai, People’s Republic of ChinaGeneral Hospital of the Tibet Military Area Command, Tibet, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University Shanghai, People’s Republic of ChinaEnterovirus 71 (EV71) is typically transmitted by the oral-faecal route and initiates infection upon crossing the intestinal mucosa. Our limited understanding of the mechanisms by which it crosses the intestinal mucosa has hampered the development of effective therapeutic options. Here, using an RNA interference screen combined with chemical inhibitors or the overexpression of dominant negative proteins, we found that EV71 entry into Caco-2 cells, a polarized human intestinal epithelial cell line, does not involve clathrin- and caveolae-dependent endocytic pathways or macropinocytosis but requires GTP-binding protein dynamin 2 and cytoskeleton remodelling. The use of siRNAs targeting endophilin family members revealed that endophlin-A2 is essential for the uptake of EV71 particles by Caco-2 cells. Subcellular analysis revealed that internalized EV71 virions largely colocalized with endophilin-A2 at cytomembrane ruffles and in the perinuclear area. Combined with viral entry kinetics, these data suggest that EV71 enters Caco-2 cells mainly via an endophilin-A2-mediated endocytic (EME) pathway. Finally, we showed that internalized EV71 virions were transported to endosomal sorting complex required for transport (ESCRT)-related multivesicular bodies (MVBs). These data provide attractive therapeutic targets to block EV71 infection.https://www.tandfonline.com/doi/10.1080/22221751.2019.1618686Enterovirus 71viral entryendophilinintestinal epithelial cellsendocytosis
spellingShingle Sheng-Lin Chen
Yan-Gang Liu
Yong-Tao Zhou
Ping Zhao
Hao Ren
Man Xiao
Yong-Zhe Zhu
Zhong-Tian Qi
Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
Emerging Microbes and Infections
Enterovirus 71
viral entry
endophilin
intestinal epithelial cells
endocytosis
title Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
title_full Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
title_fullStr Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
title_full_unstemmed Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
title_short Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
title_sort endophilin a2 mediated endocytic pathway is critical for enterovirus 71 entry into caco 2 cells
topic Enterovirus 71
viral entry
endophilin
intestinal epithelial cells
endocytosis
url https://www.tandfonline.com/doi/10.1080/22221751.2019.1618686
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