Visualization of Double-Stranded RNA Colocalizing With Pattern Recognition Receptors in Arenavirus Infected Cells

Front Cell Infect Microbiol. 2018 Jul 24:8:251. doi: 10.3389/fcimb.2018.00251. eCollection 2018.

Abstract

An important step in the initiation of the innate immune response to virus infection is the recognition of non-self, viral RNA, including double-stranded RNA (dsRNA), by cytoplasmic pattern recognition receptors (PRRs). For many positive-sense RNA viruses and DNA viruses, the production of viral dsRNA, and the interaction of viral dsRNA and PRRs are well characterized. However, for negative-sense RNA viruses, viral dsRNA was thought to be produced at low to undetectable levels and PRR recognition of viral dsRNA is still largely unclear. In the case of arenaviruses, the nucleocaspid protein (NP) has been identified to contain an exoribonuclease activity that preferentially degrades dsRNA in biochemical studies. Nevertheless, pathogenic New World (NW) arenavirus infections readily induce an interferon (IFN) response in a RIG-I dependent manner, and also activate the dsRNA-dependent Protein Kinase R (PKR). To better understand the innate immune response to pathogenic arenavirus infection, we used a newly identified dsRNA-specific antibody that efficiently detects viral dsRNA in negative-sense RNA virus infected cells. dsRNA was detected in NW arenavirus infected cells colocalizing with virus NP in immunofluorescence assay. Importantly, the dsRNA signals also colocalized with cytoplasmic PRRs, namely, PKR, RIG-I and MDA-5, as well as with the phosphorylated, activated form of PKR in infected cells. Our data clearly demonstrate the PRR recognition of dsRNA and their activation in NW arenavirus infected cells. These findings provide new insights into the interaction between NW arenaviruses and the host innate immune response.

Keywords: Junín virus; MDA-5; PKR; RIG-I; arenaviruses; dsRNA; negative-sense RNA virus; pattern recognition receptor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Arenavirus / growth & development*
  • DEAD Box Protein 58 / analysis
  • Epithelial Cells / immunology*
  • Epithelial Cells / virology*
  • Host-Pathogen Interactions*
  • Humans
  • Interferon-Induced Helicase, IFIH1 / analysis
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • RNA, Double-Stranded / analysis*
  • RNA, Viral / analysis*
  • Receptors, Immunologic
  • Receptors, Pattern Recognition / analysis*
  • eIF-2 Kinase / analysis

Substances

  • RNA, Double-Stranded
  • RNA, Viral
  • Receptors, Immunologic
  • Receptors, Pattern Recognition
  • EIF2AK2 protein, human
  • eIF-2 Kinase
  • RIGI protein, human
  • IFIH1 protein, human
  • DEAD Box Protein 58
  • Interferon-Induced Helicase, IFIH1