Influence of acenocoumarol on the interaction between Bifidobacterium and THP-1 cells

149 ASSAD, S.; FRAGOMENO, M.; MINNAARD, J. AND PÉREZ, P. F.

Congreso; XII Congreso Argentino de Microbiología General; 2017

Bifidobacteria are microorganisms widely used as probiotics. In previous studies we demonstrated the strain-dependent immunomodulatory effects on THP-1 cells, a model of phagocytic cells. It has been demonstrated that probiotics modify the bioavailability of certain drugs but there is no evidence of the effect of drugs on probiotic activity Our goal is to study the effect of acenocoumarol, a widespread oral anticoagulant, on the interaction between Bifidobacteria and THP-1 cells.THP1 cells were differentiated with phorbol miristate acetate 200 nM in DMEM (10% Fetal bovine serum) for 48h at 37ºC/5% CO2. Bifidobacterium bifidum strain CIDCA 5310 and Bifidobacterium adolescentis strain CIDCA 5317 were cultured (48h;37ºC) in MRS broth in anaerobic conditions. To evaluate the effect of the drug on the phagocytosis activity, FITC-labeled bacteria were incubated with cells at MOI 10 (multiplicity of infection) and acenocoumarol at different concentrations for 1h at 37ºC/5% CO2. Phagocytosis was evaluated by flow cytometry. For quenching of non-internalized bacteria, trypan blue was used. UI (Uptake Index) = Percentage of FL1(+) cells x mean fluorescence intensity, was calculated. Intracellular localization of bacteria was evaluated by confocal laser microscopy by using Lysotracker DND-99 and Transferrin Alexa-594 as markers of acidic and recycling compartments respectively. The expression of HLA-DR and TLR2 was measured by flow cytometry after 16hs of incubation with the strain CIDCA 5310 and the drug. Medium Fluorescence Intensity (MFI) was determined. A decrease (P