title: Exosomes released from M. tuberculosis infected cells can suppress IFN-γ mediated activation of naïve macrophages.
abstract: BACKGROUND
  Macrophages infected with Mycobacterium tuberculosis (M.tb) are known to be refractory to IFN-γ stimulation. Previous studies have shown that M.tb express components such as the 19-kDa lipoprotein and peptidoglycan that can bind to macrophage receptors including the Toll-like receptor 2 resulting in the loss in IFN-γ responsiveness. However, it is unclear whether this effect is limited to infected macrophages. We have previously shown that M.tb-infected macrophages release exosomes which are 30-100 nm membrane bound vesicles of endosomal origin that function in intercellular communication. These exosomes contain mycobacterial components including the 19-kDa lipoprotein and therefore we hypothesized that macrophages exposed to exosomes may show limited response to IFN-γ stimulation.
  
  
  RESULTS
  Exosomes were isolated from resting as well as M.tb-infected RAW264.7 macrophages. Mouse bone marrow-derived macrophages (BMMØ) were treated with exosomes +/- IFN-γ. Cells were harvested and analyzed for suppression of IFN-γ responsive genes by flow cytometry and real time PCR. We found that exosomes derived from M.tb H37Rv-infected but not from uninfected macrophages inhibited IFN-γ induced MHC class II and CD64 expression on BMMØ. This inhibition was only partially dependent on the presence of lipoproteins but completely dependent on TLR2 and MyD88. The exosomes isolated from infected cells did not inhibit STAT1 Tyrosine phosphorylation but down-regulated IFN-γ induced expression of the class II major histocompatibility complex transactivator; a key regulator of class II MHC expression. Microarray studies showed that subsets of genes induced by IFN-γ were inhibited by exosomes from H37Rv-infected cells including genes involved in antigen presentation. Moreover, this set of genes partially overlapped with the IFN-γ-induced genes inhibited by H37Rv infection.
  
  
  CONCLUSIONS
  Our study suggests that exosomes, as carriers of M.tb pathogen associated molecular patterns (PAMPs), may provide a mechanism by which M.tb may exert its suppression of a host immune response beyond the infected cell.
authors.affiliation: Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana, United States of America.
authors.forename: Prachi P
authors.initials: PP
authors.lastname: Singh
authors.forename: Christopher
authors.initials: C
authors.lastname: LeMaire
authors.forename: John C
authors.initials: JC
authors.lastname: Tan
authors.forename: Erliang
authors.initials: E
authors.lastname: Zeng
authors.forename: Jeffery S
authors.initials: JS
authors.lastname: Schorey
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D055354:Exosomes; D005260:Female; D005434:Flow Cytometry; D007371:Interferon-gamma / Q000502:physiology*; D008262:Macrophage Activation / Q000502:physiology*; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000502:physiology*; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D050794:STAT1 Transcription Factor / Q000378:metabolism
chemical_list: D017931:DNA Primers; D050794:STAT1 Transcription Factor; C494080:Stat1 protein, mouse; D007371:Interferon-gamma