ABSTRACT
Oxidized cholesterols have emerged as important signaling molecules of immune function, but little is known about the role of these oxysterols during mycobacterial infections. We found that expression of the oxysterol-receptor GPR183 was reduced in blood from patients with tuberculosis (TB) and type 2 diabetes (T2D) compared to TB patients without T2D and was associated with TB disease severity on chest x-ray. GPR183 activation by 7α,25-dihydroxycholesterol (7α,25-OHC) reduced growth of Mycobacterium tuberculosis (Mtb) and Mycobacterium bovis BCG in primary human monocytes, an effect abrogated by the GPR183 antagonist GSK682753. Growth inhibition was associated with reduced IFN-ß and IL-10 expression and enhanced autophagy. Mice lacking GPR183 had significantly increased lung Mtb burden and dysregulated IFNs during early infection. Together, our data demonstrate that GPR183 is an important regulator of intracellular mycobacterial growth and interferons during mycobacterial infection.
Subject(s)
Autophagy , Interferons/metabolism , Leukocytes, Mononuclear/microbiology , Lung/microbiology , Mycobacterium tuberculosis/growth & development , Receptors, G-Protein-Coupled/metabolism , Tuberculosis, Pulmonary/microbiology , Animals , Bacterial Load , Case-Control Studies , Diabetes Mellitus, Type 2/immunology , Diabetes Mellitus, Type 2/metabolism , Disease Models, Animal , Female , Host-Pathogen Interactions , Humans , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Lung/immunology , Lung/metabolism , Male , Mice, Inbred C57BL , Mice, Knockout , Mycobacterium bovis/growth & development , Mycobacterium bovis/immunology , Mycobacterium bovis/pathogenicity , Mycobacterium tuberculosis/immunology , Mycobacterium tuberculosis/pathogenicity , Receptors, G-Protein-Coupled/genetics , Severity of Illness Index , Signal Transduction , THP-1 Cells , Tuberculosis, Pulmonary/immunology , Tuberculosis, Pulmonary/metabolismABSTRACT
The emergence of antibiotic-resistant bacterial pathogens, especially Gram-negative bacteria, has driven investigations into suppressing bacterial virulence via quorum sensing (QS) inhibition strategies instead of bactericidal and bacteriostatic approaches. Here, we investigated several bee products for potential compound(s) that exhibit significant QS inhibitory (QSI) properties at the phenotypic and molecular levels in Chromobacterium violaceum ATCC 12472 as a model organism. Manuka propolis produced the strongest violacein inhibition on C. violaceum lawn agar, while bee pollen had no detectable QSI activity and honey had bactericidal activity. Fractionated manuka propolis (pooled fraction 5 or PF5) exhibited the largest violacein inhibition zone (24.5 ± 2.5 mm) at 1 mg dry weight per disc. In C. violaceum liquid cultures, at least 450 µg/ml of manuka propolis PF5 completely inhibited violacein production. Gene expression studies of the vioABCDE operon, involved in violacein biosynthesis, showed significant (≥two-fold) down-regulation of vioA, vioD and vioE in response to manuka propolis PF5. A potential QSI compound identified in manuka propolis PF5 is a hydroxycinnamic acid-derivative, isoprenyl caffeate, with a [M-H] of 247. Complete violacein inhibition in C. violaceum liquid cultures was achieved with at least 50 µg/ml of commercial isoprenyl caffeate. In silico docking experiments suggest that isoprenyl caffeate may act as an inhibitor of the violacein biosynthetic pathway by acting as a competitor for the FAD-binding pockets of VioD and VioA. Further studies on these compounds are warranted toward the development of anti-pathogenic drugs as adjuvants to conventional antibiotic treatments, especially in antibiotic-resistant bacterial infections.
