Protein kinase R mediates intestinal epithelial gene remodeling in response to double-stranded RNA and live rotavirus

Matam Vijay-Kumar, Jon R. Gentsch, William J. Kaiser, Niels Borregaard, Margaret K. Offermann, Andrew S. Neish, Andrew T. Gewirtz

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40 Scopus citations

Abstract

As sentinels of host defense, intestinal epithelial cells respond to the viral pathogen rotavirus by activating a gene expression that promotes immune cell recruitment and activation. We hypothesized that epithelial sensing of rotavirus might target dsRNA, which can be detected by TLR3 or protein kinase R (PKR). Accordingly, we observed that synthetic dsRNA, polyinosinic acid:cytidylic acid (poly(I:C)), potently induced gene remodeling in model intestinal epithelia with the specific pattern of expressed genes, including both classic proinflammatory genes (e.g., IL-8), as well as genes that are classically activated in virus-infected cells (e.g., IFN-responsive genes). Poly(I:C)-induced IL-8 was concentration dependent (2-100 μg/ml) and displayed slower kinetics compared with IL-8 induced by bacterial flagellin (ET50 ∼24 vs 8 h poly(I:C) vs flagellin, respectively). Although model epithelia expressed detectable TLR3 mRNA, neither TLR3-neutralizing Abs nor chloroquine, which blocks activation of intracellular TLR3, attenuated epithelial responses to poly(I:C). Conversely, poly(I:C)-induced phosphorylation of PKR and inhibitors of PKR, 2-aminopurine and adenine, ablated poly(I:C)-induced gene expression but had no effect on gene expression induced by flagellin, thus suggesting that intestinal epithelial cell detection of dsRNA relies on PKR. Consistent with poly(I:C) detection by an intracellular molecule such as PKR, we observed that both uptake of and responses to poly(I:C) were polarized to the basolateral side. Lastly, we observed that the pattern of pharmacologic inhibition of responses to poly(I:C) was identical to that seen in response to infection by live rotavirus, indicating a potentially important role for PKR in activating intestinal epithelial gene expression in rotavirus infection.

Original languageEnglish (US)
Pages (from-to)6322-6331
Number of pages10
JournalJournal of Immunology
Volume174
Issue number10
DOIs
StatePublished - May 15 2005

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ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Vijay-Kumar, M., Gentsch, J. R., Kaiser, W. J., Borregaard, N., Offermann, M. K., Neish, A. S., & Gewirtz, A. T. (2005). Protein kinase R mediates intestinal epithelial gene remodeling in response to double-stranded RNA and live rotavirus. Journal of Immunology, 174(10), 6322-6331. https://doi.org/10.4049/jimmunol.174.10.6322