Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection

ABSTRACT Human noroviruses (HuNoVs) are the leading cause of nonbacterial gastroenteritis worldwide. Histo-blood group antigen (HBGA) expression is an important susceptibility factor for HuNoV infection based on controlled human infection models and epidemiologic studies that show an association of...

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Autores principales:	Kei Haga, Khalil Ettayebi, Victoria R. Tenge, Umesh C. Karandikar, Miranda A. Lewis, Shih-Ching Lin, Frederick H. Neill, B. Vijayalakshmi Ayyar, Xi-Lei Zeng, Göran Larson, Sasirekha Ramani, Robert L. Atmar, Mary K. Estes
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	fucosyltransferase 2 glycobiology histo-blood group antigens isogenic enteroids isogenic organoids noroviruses Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c7d23fae05e64407a2dfc2249cda3572
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spelling	oai:doaj.org-article:c7d23fae05e64407a2dfc2249cda35722021-11-15T15:57:01ZGenetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection10.1128/mBio.00251-202150-7511https://doaj.org/article/c7d23fae05e64407a2dfc2249cda35722020-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00251-20https://doaj.org/toc/2150-7511ABSTRACT Human noroviruses (HuNoVs) are the leading cause of nonbacterial gastroenteritis worldwide. Histo-blood group antigen (HBGA) expression is an important susceptibility factor for HuNoV infection based on controlled human infection models and epidemiologic studies that show an association of secretor status with infection caused by several genotypes. The fucosyltransferase 2 gene (FUT2) affects HBGA expression in intestinal epithelial cells; secretors express a functional FUT2 enzyme, while nonsecretors lack this enzyme and are highly resistant to infection and gastroenteritis caused by many HuNoV strains. These epidemiologic associations are confirmed by infections in stem cell-derived human intestinal enteroid (HIE) cultures. GII.4 HuNoV does not replicate in HIE cultures derived from nonsecretor individuals, while HIEs from secretors are permissive to infection. However, whether FUT2 expression alone is critical for infection remains unproven, since routinely used secretor-positive transformed cell lines are resistant to HuNoV replication. To evaluate the role of FUT2 in HuNoV replication, we used CRISPR or overexpression to genetically manipulate FUT2 gene function to produce isogenic HIE lines with or without FUT2 expression. We show that FUT2 expression alone affects both HuNoV binding to the HIE cell surface and susceptibility to HuNoV infection. These findings indicate that initial binding to a molecule(s) glycosylated by FUT2 is critical for HuNoV infection and that the HuNoV receptor is present in nonsecretor HIEs. In addition to HuNoV studies, these isogenic HIE lines will be useful tools to study other enteric microbes where infection and/or disease outcome is associated with secretor status. IMPORTANCE Several studies have demonstrated that secretor status is associated with susceptibility to human norovirus (HuNoV) infection; however, previous reports found that FUT2 expression is not sufficient to allow infection with HuNoV in a variety of continuous laboratory cell lines. Which cellular factor(s) regulates susceptibility to HuNoV infection remains unknown. We used genetic manipulation of HIE cultures to show that secretor status determined by FUT2 gene expression is necessary and sufficient to support HuNoV replication based on analyses of isogenic lines that lack or express FUT2. Fucosylation of HBGAs is critical for initial binding and for modification of another putative receptor(s) in HIEs needed for virus uptake or uncoating and necessary for successful infection by GI.1 and several GII HuNoV strains.Kei HagaKhalil EttayebiVictoria R. TengeUmesh C. KarandikarMiranda A. LewisShih-Ching LinFrederick H. NeillB. Vijayalakshmi AyyarXi-Lei ZengGöran LarsonSasirekha RamaniRobert L. AtmarMary K. EstesAmerican Society for Microbiologyarticlefucosyltransferase 2glycobiologyhisto-blood group antigensisogenic enteroidsisogenic organoidsnorovirusesMicrobiologyQR1-502ENmBio, Vol 11, Iss 2 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	fucosyltransferase 2 glycobiology histo-blood group antigens isogenic enteroids isogenic organoids noroviruses Microbiology QR1-502
spellingShingle	fucosyltransferase 2 glycobiology histo-blood group antigens isogenic enteroids isogenic organoids noroviruses Microbiology QR1-502 Kei Haga Khalil Ettayebi Victoria R. Tenge Umesh C. Karandikar Miranda A. Lewis Shih-Ching Lin Frederick H. Neill B. Vijayalakshmi Ayyar Xi-Lei Zeng Göran Larson Sasirekha Ramani Robert L. Atmar Mary K. Estes Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection
description	ABSTRACT Human noroviruses (HuNoVs) are the leading cause of nonbacterial gastroenteritis worldwide. Histo-blood group antigen (HBGA) expression is an important susceptibility factor for HuNoV infection based on controlled human infection models and epidemiologic studies that show an association of secretor status with infection caused by several genotypes. The fucosyltransferase 2 gene (FUT2) affects HBGA expression in intestinal epithelial cells; secretors express a functional FUT2 enzyme, while nonsecretors lack this enzyme and are highly resistant to infection and gastroenteritis caused by many HuNoV strains. These epidemiologic associations are confirmed by infections in stem cell-derived human intestinal enteroid (HIE) cultures. GII.4 HuNoV does not replicate in HIE cultures derived from nonsecretor individuals, while HIEs from secretors are permissive to infection. However, whether FUT2 expression alone is critical for infection remains unproven, since routinely used secretor-positive transformed cell lines are resistant to HuNoV replication. To evaluate the role of FUT2 in HuNoV replication, we used CRISPR or overexpression to genetically manipulate FUT2 gene function to produce isogenic HIE lines with or without FUT2 expression. We show that FUT2 expression alone affects both HuNoV binding to the HIE cell surface and susceptibility to HuNoV infection. These findings indicate that initial binding to a molecule(s) glycosylated by FUT2 is critical for HuNoV infection and that the HuNoV receptor is present in nonsecretor HIEs. In addition to HuNoV studies, these isogenic HIE lines will be useful tools to study other enteric microbes where infection and/or disease outcome is associated with secretor status. IMPORTANCE Several studies have demonstrated that secretor status is associated with susceptibility to human norovirus (HuNoV) infection; however, previous reports found that FUT2 expression is not sufficient to allow infection with HuNoV in a variety of continuous laboratory cell lines. Which cellular factor(s) regulates susceptibility to HuNoV infection remains unknown. We used genetic manipulation of HIE cultures to show that secretor status determined by FUT2 gene expression is necessary and sufficient to support HuNoV replication based on analyses of isogenic lines that lack or express FUT2. Fucosylation of HBGAs is critical for initial binding and for modification of another putative receptor(s) in HIEs needed for virus uptake or uncoating and necessary for successful infection by GI.1 and several GII HuNoV strains.
format	article
author	Kei Haga Khalil Ettayebi Victoria R. Tenge Umesh C. Karandikar Miranda A. Lewis Shih-Ching Lin Frederick H. Neill B. Vijayalakshmi Ayyar Xi-Lei Zeng Göran Larson Sasirekha Ramani Robert L. Atmar Mary K. Estes
author_facet	Kei Haga Khalil Ettayebi Victoria R. Tenge Umesh C. Karandikar Miranda A. Lewis Shih-Ching Lin Frederick H. Neill B. Vijayalakshmi Ayyar Xi-Lei Zeng Göran Larson Sasirekha Ramani Robert L. Atmar Mary K. Estes
author_sort	Kei Haga
title	Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection
title_short	Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection
title_full	Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection
title_fullStr	Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection
title_full_unstemmed	Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection
title_sort	genetic manipulation of human intestinal enteroids demonstrates the necessity of a functional fucosyltransferase 2 gene for secretor-dependent human norovirus infection
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/c7d23fae05e64407a2dfc2249cda3572
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Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection

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