TLR7 is a key regulator of innate immunity against Japanese encephalitis virus infection

Neurobiol Dis. 2014 Sep:69:235-47. doi: 10.1016/j.nbd.2014.05.036. Epub 2014 Jun 5.

Abstract

Toll-like receptor 7 (TLR7) known to recognize guanidine-rich ssRNA has been shown to mount vital host defense mechanism against many viruses including flaviviruses. Signal transduction through TLR7 has been shown to produce type-1 interferon and proinflammatory mediators, thereby initiating essential innate immune response against ssRNA viruses in hosts. Systemic and brain specific TLR7 knock-down mice (TLR7(KD)) were generated using vivo-morpholinos. These mice were then subcutaneously challenged with lethal dose of JEV (GP78 strain) and were subsequently analyzed for survival. Significant difference in susceptibility to JEV between wild-type and systemic TLR7(KD) mice was observed whereas, no difference in susceptibility to JEV infection was seen in brain-specific TLR7(KD) mice. Significant decreases in IFN-α and antiviral proteins were also observed in both TLR7(KD) mice along with increased viral loads in their brain. Owing to increased viral load, increases in levels of various proinflammatory cyto/chemokines, increased microglial activation and infiltration of peripheral immune cells in brain of TLR7(KD) mice were also observed. Immunocytochemistry and RNA co-immunoprecipitation performed with JEV-infected N2a or HT22 cells indicated endosomal localization and confirmed interaction between JEV ssRNA with TLR7. Treatment of mice with imiquimod, a TLR7 agonist, prior to JEV infection resulted in their increased survival. Overall, our results suggest that the TLR7 response following JEV infection promotes type-1 interferon production and generation of antiviral state which might contribute to protective effect in systemic infection.

Keywords: Antiviral; Innate immunity; Japanese encephalitis; Neurons; Neurotrophic virus; Pattern recognition receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / pharmacology
  • Aminoquinolines / pharmacology
  • Animals
  • Brain / immunology
  • Brain / virology
  • Cell Line
  • Cell Line, Tumor
  • Cells, Cultured
  • DNA, Viral / metabolism
  • Disease Models, Animal
  • Encephalitis Virus, Japanese / genetics
  • Encephalitis, Japanese / immunology*
  • Encephalitis, Japanese / prevention & control
  • Encephalitis, Japanese / virology
  • Gene Knockdown Techniques
  • Humans
  • Imiquimod
  • Immunity, Innate*
  • Interferon Type I / metabolism
  • Membrane Glycoproteins / agonists
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism*
  • Mice, Inbred BALB C
  • Mice, Transgenic
  • Microglia / physiology
  • Neurons / immunology
  • Toll-Like Receptor 7 / agonists
  • Toll-Like Receptor 7 / genetics
  • Toll-Like Receptor 7 / metabolism*

Substances

  • Adjuvants, Immunologic
  • Aminoquinolines
  • DNA, Viral
  • Interferon Type I
  • Membrane Glycoproteins
  • TLR7 protein, human
  • Tlr7 protein, mouse
  • Toll-Like Receptor 7
  • Imiquimod