B. abortus RNA is the component involved in the down-modulation of MHC-I expression on human monocytes via TLR8 and the EGFR pathway

PLoS Pathog. 2017 Aug 2;13(8):e1006527. doi: 10.1371/journal.ppat.1006527. eCollection 2017 Aug.

Abstract

Despite eliciting a potent CD8+ T cell response, Brucella abortus is able to persist and establish a chronic infection inside its host. We have previously reported that the infection of human monocytes/macrophages with B. abortus inhibits the IFN-γ-induced MHC-I cell surface expression down-modulating cytotoxic CD8+ T cell responses. MHC-I down-modulation depends on bacterial viability and results from the capacity of B. abortus to retain the MHC-I molecules within the Golgi apparatus. Furthermore, we recently demonstrated that epidermal growth factor receptor (EGFR) pathway is involved in this phenomenon and that this is an early event during infection. However, the components and mechanisms whereby B. abortus is able to down-modulate MHC-I remained to be elucidated. In this study we demonstrated that the down-modulation of MHC-I expression is not mediated by well-known Brucella virulence factors but instead by B. abortus RNA, a PAMP associated to viability (vita-PAMP). Surprisingly, completely degraded RNA was also able to inhibit MHC-I expression to the same extent as intact RNA. Accordingly, B. abortus RNA and its degradation products were able to mimic the MHC-I intracellular retention within the Golgi apparatus observed upon infection. We further demonstrated that TLR8, a single-stranded RNA and RNA degradation products sensor, was involved in MHC-I inhibition. On the other hand, neutralization of the EGFR reversed the MHC-I inhibition, suggesting a connection between the TLR8 and EGFR pathways. Finally, B. abortus RNA-treated macrophages display diminished capacity of antigen presentation to CD8+ T cells. Overall, our results indicate that the vita-PAMP RNA as well as its degradation products constitute novel virulence factors whereby B. abortus, by a TLR8-dependent mechanism and through the EGFR pathway, inhibits the IFN-γ-induced MHC-I surface expression on human monocytes/macrophages. Thus, bacteria can hide within infected cells and avoid the immunological surveillance of cytotoxic CD8+ T cells.

MeSH terms

  • Animals
  • Brucella abortus / immunology
  • Brucellosis / immunology*
  • Cross-Priming / immunology
  • Down-Regulation
  • Enzyme-Linked Immunosorbent Assay
  • ErbB Receptors / immunology*
  • Flow Cytometry
  • Histocompatibility Antigens Class I / biosynthesis
  • Humans
  • Immune Evasion / immunology*
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Confocal
  • Monocytes / immunology*
  • Monocytes / microbiology
  • RNA, Bacterial / immunology*
  • Signal Transduction / immunology
  • Toll-Like Receptor 8 / immunology*

Substances

  • Histocompatibility Antigens Class I
  • RNA, Bacterial
  • TLR8 protein, human
  • Toll-Like Receptor 8
  • EGFR protein, human
  • ErbB Receptors

Grants and funding

This work was supported by grants PICT 2011-1420 (PB), 2011-1200 (GHG), 2012-2252 (GHG), 2013-0162 (PB) and 2014-1925 (GHG) from the Agencia Nacional de Promoción Científica y Tecnológica (ANPCYT-Argentina), by grants PIP 1390 (PB) and PIP 0373 (PB) from Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET-Argentina), by grants UBACYT 20020130200030 (PB) and 20020120100128 (GHG) from the University of Buenos Aires, and by grant from Fundación Alberto J. Roemmers (2012-2014) (PB) (Argentina). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.