Brucella abortus Triggers a cGAS-Independent STING Pathway To Induce Host Protection That Involves Guanylate-Binding Proteins and Inflammasome Activation

J Immunol. 2018 Jan 15;200(2):607-622. doi: 10.4049/jimmunol.1700725. Epub 2017 Dec 4.

Abstract

Immunity against microbes depends on recognition of pathogen-associated molecular patterns by innate receptors. Signaling pathways triggered by Brucella abortus DNA involves TLR9, AIM2, and stimulator of IFN genes (STING). In this study, we observed by microarray analysis that several type I IFN-associated genes, such as IFN-β and guanylate-binding proteins (GBPs), are downregulated in STING knockout (KO) macrophages infected with Brucella or transfected with DNA. Additionally, we determined that STING and cyclic GMP-AMP synthase (cGAS) are important to engage the type I IFN pathway, but only STING is required to induce IL-1β secretion, caspase-1 activation, and GBP2 and GBP3 expression. Furthermore, we determined that STING but not cGAS is critical for host protection against Brucella infection in macrophages and in vivo. This study provides evidence of a cGAS-independent mechanism of STING-mediated protection against an intracellular bacterial infection. Additionally, infected IFN regulatory factor-1 and IFNAR KO macrophages had reduced GBP2 and GBP3 expression and these cells were more permissive to Brucella replication compared with wild-type control macrophages. Because GBPs are critical to target vacuolar bacteria, we determined whether GBP2 and GBPchr3 affect Brucella control in vivo. GBPchr3 but not GBP2 KO mice were more susceptible to bacterial infection, and small interfering RNA treated-macrophages showed reduction in IL-1β secretion and caspase-1 activation. Finally, we also demonstrated that Brucella DNA colocalizes with AIM2, and AIM2 KO mice are less resistant to B. abortus infection. In conclusion, these findings suggest that the STING-dependent type I IFN pathway is critical for the GBP-mediated release of Brucella DNA into the cytosol and subsequent activation of AIM2.

Publication types

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

MeSH terms

  • Animals
  • Brucella abortus / genetics
  • Brucella abortus / immunology*
  • Brucellosis / genetics
  • Brucellosis / immunology*
  • Brucellosis / metabolism*
  • Brucellosis / microbiology
  • Cyclic GMP / analogs & derivatives
  • Cyclic GMP / metabolism
  • Cytokines / metabolism
  • GTP-Binding Proteins / genetics
  • GTP-Binding Proteins / metabolism*
  • Gene Expression
  • Gene Expression Profiling
  • Granuloma / metabolism
  • Granuloma / microbiology
  • Granuloma / pathology
  • Host-Pathogen Interactions / genetics
  • Host-Pathogen Interactions / immunology
  • Immunity, Innate
  • Inflammasomes / metabolism*
  • Inflammation Mediators
  • Interferon Regulatory Factor-3 / metabolism
  • Interferon Type I / genetics
  • Interferon Type I / metabolism
  • Macrophages / immunology
  • Macrophages / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mice
  • Mice, Knockout
  • Models, Biological
  • NF-kappa B / metabolism
  • Signal Transduction*

Substances

  • Cytokines
  • Inflammasomes
  • Inflammation Mediators
  • Interferon Regulatory Factor-3
  • Interferon Type I
  • Membrane Proteins
  • NF-kappa B
  • Sting1 protein, mouse
  • bis(3',5')-cyclic diguanylic acid
  • GTP-Binding Proteins
  • Cyclic GMP