Targeted Profiling of Immunological Genes during Norovirus Replication in Human Intestinal Enteroids

Viruses. 2021 Jan 21;13(2):155. doi: 10.3390/v13020155.

Abstract

Norovirus is the leading cause of acute gastroenteritis worldwide. The pathogenesis of norovirus and the induced immune response remain poorly understood due to the lack of a robust virus culture system. The monolayers of two secretor-positive Chinese human intestinal enteroid (HIE) lines were challenged with two norovirus pandemic GII.4 Sydney strains. Norovirus RNA replication in supernatants and cell lysates were quantified by RT-qPCR. RNA expression levels of immune-related genes were profiled using PCR arrays. The secreted protein levels of shortlisted upregulated genes were measured in supernatants using analyte-specific enzyme-linked immunosorbent assay (ELISA). Productive norovirus replications were achieved in three (75%) out of four inoculations. The two most upregulated immune-related genes were CXCL10 (93-folds) and IFI44L (580-folds). Gene expressions of CXCL10 and IFI44L were positively correlated with the level of norovirus RNA replication (CXCL10: Spearman's r = 0.779, p < 0.05; IFI44L: r = 0.881, p < 0.01). The higher level of secreted CXCL10 and IFI44L proteins confirmed their elevated gene expression. The two genes have been reported to be upregulated in norovirus volunteer challenges and natural human infections by other viruses. Our data suggested that HIE could mimic the innate immune response elicited in natural norovirus infection and, therefore, could serve as an experimental model for future virus-host interaction and antiviral studies.

Keywords: human intestinal enteroids; human norovirus; innate immune response; virus–host interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Caliciviridae Infections / immunology*
  • Cell Line
  • Chemokine CXCL10 / genetics
  • Chemokine CXCL10 / metabolism*
  • Female
  • Host Microbial Interactions
  • Humans
  • Immunity, Innate
  • Interferons / genetics
  • Interferons / metabolism
  • Intestines / immunology
  • Intestines / virology*
  • Male
  • Middle Aged
  • Models, Biological
  • Norovirus / pathogenicity
  • Norovirus / physiology
  • Organoids / immunology
  • Organoids / virology
  • Sequence Analysis, RNA
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / metabolism*
  • Virus Replication

Substances

  • CXCL10 protein, human
  • Chemokine CXCL10
  • IFI44L protein, human
  • Tumor Suppressor Proteins
  • Interferons