Human Norovirus Propagation in Human Induced Pluripotent Stem Cell–Derived Intestinal Epithelial Cells

Sato, Shintaro; Hisaie, Kota; Kurokawa, Shiho; Suzuki, Akio; Sakon, Naomi; Uchida, Yohei; Yuki, Yoshikazu; Kiyono, Hiroshi

doi:10.1016/j.jcmgh.2018.11.001

Cited by 54 publications

(57 citation statements)

References 10 publications

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“…We further examined the morphological change in virus particles after the incubation with disinfectants. To this end, we used virus-like particles (VLPs), which comprise the VP1 capsid proteins of GII.17 4 . Negative stain transmission electron microscopy analysis revealed that 0.1% NaClO could burst VLPs (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In 2016, Estes and her colleagues first reported that several HuNoV genotypes could propagate in IECs prepared from human small intestines 3 . Thereafter, our group also reported that human iPSC-derived IECs could be used for the cultivation of HuNoVs 4 . In this study, we investigated whether alcohol-based disinfectants have actual virucidal effects on several genotypes of HuNoV using our in vitro HuNoV cultivation system by analyzing its propagative potency as an index.…”

Section: Discussionmentioning

confidence: 97%

“…The differentiation of human iPSCs into IECs and their culture were performed as described previously 4 , 20 . Each conditioned medium (CM) was prepared as described previously 20 .…”

Section: Methodsmentioning

confidence: 99%

“…One of the reasons for this is that there were no in vitro cultivation systems for HuNoVs until 2016 3 . The recent development of a HuNoV replication system in human primary intestinal epithelial cells (IECs) and human induced pluripotent stem cell (iPSC)-derived IECs has spawned advances in HuNoV characterization 4 . Until then, the inactivation methods of HuNoVs were determined using feline calicivirus and/or murine norovirus as surrogates 5 – 7 .…”

Section: Introductionmentioning

confidence: 99%

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Alcohol abrogates human norovirus infectivity in a pH-dependent manner

Sato

Matsumoto

Hisaie

et al. 2020

Sci Rep

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Alcohol-based disinfectants are widely used for the sanitization of microorganisms, especially those that cause infectious diseases, including viruses. However, since the germicidal mechanism of alcohol is lipolysis, alcohol-based disinfectants appear to have a minimal effect on non-enveloped viruses, such as noroviruses. Because there is no cultivation method for human norovirus (HuNoV) in vitro, murine norovirus and feline calicivirus have been used as surrogates for HuNoV to analyze the efficacy of disinfectant regents. Therefore, whether these disinfectants and their conditions are effective against HuNoVs remain unknown. In this study, we report that ethanol or isopropanol alone can sufficiently suppress GII.4 genotype HuNoV replication in human iPSC-derived intestinal epithelial cells. Additionally, pH adjustments and salting-out may contribute toward the virucidal effect of alcohol against other HuNoV genotypes and cancel the impediment of organic substance contamination, respectively. Therefore, similar to sodium hypochlorite, alcohol-based disinfectants containing electrolytes can be used for HuNoV inactivation.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 97%

“…The differentiation of human iPSCs into IECs and their culture were performed as described previously 4 , 20 . Each conditioned medium (CM) was prepared as described previously 20 .…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Alcohol abrogates human norovirus infectivity in a pH-dependent manner

Sato

Matsumoto

Hisaie

et al. 2020

Sci Rep

Self Cite

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“…We will refer to those as human intestinal enteroids (HIE) throughout. In recent years, HIE are increasingly being used as disease models for the study of host-pathogen interactions, especially in the context of infection with viruses that lack a robust cell system such as rotaviruses and noroviruses [7][8][9][10][11]. HIE are grown in extracellular matrix scaffolds in 3D and maintain aspects of their in vivo physiology for long periods in culture [12].…”

Section: Introductionmentioning

confidence: 99%

Astrovirus replication in human intestinal enteroids reveals multi-cellular tropism and an intricate host innate immune landscape

et al. 2019

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Human astroviruses (HAstV) are understudied positive-strand RNA viruses that cause gastroenteritis mostly in children and the elderly. Three clades of astroviruses, classic, MLBtype and VA-type have been reported in humans. One limitation towards a better understanding of these viruses has been the lack of a physiologically relevant cell culture model that supports growth of all clades of HAstV. Herein, we demonstrate infection of HAstV strains belonging to all three clades in epithelium-only human intestinal enteroids (HIE) isolated from biopsy-derived intestinal crypts. A detailed investigation of infection of VA1, a member of the non-canonical HAstV-VA/HMO clade, showed robust replication in HIE derived from different patients and from different intestinal regions independent of the cellular differentiation status. Flow cytometry and immunofluorescence analysis revealed that VA1 infects several cell types, including intestinal progenitor cells and mature enterocytes, in HIE cultures. RNA profiling of VA1-infected HIE uncovered that the host response to infection is dominated by interferon (IFN)-mediated innate immune responses. A comparison of the antiviral host response in non-transformed HIE and transformed human colon carcinoma Caco-2 cells highlighted significant differences between these cells, including an increased magnitude of the response in HIE. Additional studies confirmed the sensitivity of VA1 to exogenous IFNs, and indicated that the endogenous IFN response of HIE to curtail the growth of strains from all three clades. Genotypic variation in the permissiveness of different HIE lines to HAstV could be overcome by pharmacologic inhibition of JAK/STAT PLOS Pathogens | https://doi.and ALSAC to SSC.; NIH R21 NS101371 to DW; Wellcome Trust: Ref 207498/Z/ signaling. Collectively, our data identify HIE as a universal infection model for HAstV and an improved model of the intestinal epithelium to investigate enteric virus-host interactions. Author summaryHuman astroviruses (HAstV) are understudied positive-strand RNA viruses that typically cause gastroenteritis mostly in children and the elderly, but more recent studies also implicate them in neurological disease in immunocompromised patients. To better understand these viruses, a physiologically relevant cell culture model that supports growth of all clades of HAstV would be highly beneficial. Herein, we demonstrated robust infection of HAstV strains belonging to all three clades in epithelium-only human intestinal enteroids (HIE) isolated from biopsy-derived intestinal crypts from different patients and intestinal regions, making HIE a valuable model to study HAstV biology. Using this system, we identify for the first time that VA1 infects several cell types, including intestinal progenitor cells and mature enterocytes. Analysis of the antiviral host response to infection demonstrated that HIE respond to infection with a type I and III interferon response. This response reduced HAstV replication and when blocked resulted in increased infection. Establis...

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