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Related Experiment Video

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Human Norovirus Efficiently Replicates in Differentiated 3D-Human Intestinal Enteroids.

Carmen Mirabelli1,2, Nanci Santos-Ferreira3, Merritt G Gillilland4

  • 1Institute for Virology and Cell Biology, University of Lübeck, Germany.

Journal of Virology
|November 7, 2022
PubMed
Summary

A new, simplified protocol for infecting 3D human intestinal enteroids (HIE) with human norovirus (HNoV) allows for faster, more effective viral studies. This model aids in developing new antiviral treatments for HNoV gastroenteritis.

Keywords:
3D-human intestinal enteroidsRNA sequencinghuman noroviruspolarity inversion

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Area of Science:

  • Virology and Gastroenterology
  • Infectious Diseases
  • Cell Biology and Host-Pathogen Interactions

Background:

  • Human norovirus (HNoV) causes significant global gastroenteritis and economic losses.
  • Current HNoV research is hampered by the lack of efficient cultivation systems.
  • Existing models using biopsy-derived human intestinal enteroids (HIE) are labor-intensive and time-consuming.

Purpose of the Study:

  • To develop a simplified and robust protocol for HNoV infection in 3D HIE.
  • To establish a more efficient platform for studying HNoV and screening antiviral agents.
  • To investigate the host transcriptional response to HNoV infection in a physiologically relevant model.

Main Methods:

  • Developed an alternative protocol for HNoV infection directly in 3D HIE.
  • Assessed differentiation efficiency and spontaneous polarity inversion using UEA-1 staining.
  • Quantified viral load and tested antiviral efficacy (e.g., 2CMC, ruxolitinib) and performed RNA sequencing.

Main Results:

  • The new protocol achieved efficient differentiation of 3D HIE, with ~80% spontaneous polarity inversion.
  • HNoV GII.4 infection showed a ~2 Log10 increase over inoculum, enhanced to 3.5 Log10 with ruxolitinib.
  • Antiviral treatments reduced HNoV infection, and RNA sequencing revealed altered host pathways (upregulated hormone/neurotransmitter signaling, downregulated glycolysis/hypoxia).

Conclusions:

  • The simplified 3D HIE model is a robust platform for HNoV infection studies and antiviral screening.
  • This model offers advantages in experimental time, infection yield, and preservation of intestinal epithelium integrity.
  • The model is suitable for studying other apical-infecting pathogens and host-microbiome interactions.