Lactobacillus reuteri CRL1098 soluble factors attenuate inflammatory response of Toll-like receptor 4-activated macrophages

GRIET M; VILLENA J; TOMASADA Y; SALVA S; FONT DE VALDEZ G; KITAZAWA H; RODRIGUEZ AV

San Miguel de Tucumán

Simposio; IV International Symposium on Lactic Acid Bacteria - Food, Health and Applications; 2013

Centro de Referencia para Lactobacilos. CERELA-CONICET

The immunomodulatory potential of probiotic bacteria is increasingly of interest for prophylactic and therapeutic options in various complex disorders including inflammatory diseases. Previously, we demonstrated that Lactobacillus reuteri CRL1098 soluble factors (LrS) are able to reduce TNF-α production in peripheral blood mononuclear cells, indicating that LrS has a potential anti-inflammatory effect. In the present work we aimed to confirm the immunoregulatory effect of LrS in mouse peritoneal macrophages under non-inflammatory conditions and after the challenge with lipopolysaccharide (LPS). Peritoneal macrophages cultures were in vitro stimulated with different concentrations LrS during 12, 24 or 48 hours and the production of TNF-α, IL-1β, IL-6, IL-12 and IL-10 was determined. We found that LrS is able to significantly decrease TNF-α production and increase IL-10 levels in a dose- and time-dependent manner. Stimulation with LrS diluted ½ during 24 hours was the optimal dose/time to achieve the immunoregulatory effect and was used for further experiments. We next evaluated whether LrS was able to modulate the response of macrophages to LPS challenge. Two series of experiments were design: a) peritoneal macrophages where stimulated with LrS in the optimal dose and then challenged with LPS (preventive treatment) or; b) macrophages were stimulated with a coadministration of LPS plus LrS. Challenge of macrophages with LPS significantly increased the production of the pro-inflammatory cytokines TNF-α, IL-1β and IL-6 (p