Development of immunobiotic evaluation system on porcine intestinal epitheliocytes stimulated by dextran sodium sulfate

NANA SATO; ASUKA TADA; WAKAKO IKEDA-OHTSUBO; TADAO SAITO; HUMAYUN KOBER; HIKARU IIDA; TOMONORI NOCHI; JULIO VILLENA; HISAKAZU KOBAYASHI; YUKI MASUMIZU; HISASHI ASO; HARUKI KITAZAWA

San Miguel de Tucuman

Simposio; V International Symposium on Lactic Acid Bacteria: Benefitting from Lactic Acid Bacteria - Progress in Health and Food; 2016

Reference Centre for Lactobacilli (CERELA-CONICET)

Previously we demonstrated that porcine intestinal epithelial (PIE) cells are a useful in vitro tool for the selection and study of immunomodulatory lactic acid bacteria (immunobiotic LAB) able to regulate the inflammatory response induced by TLR4 activation. Dextran sulphate sodium (DSS) induced colitis is well-established animal model of mucosal inflammation that has been used for decades in the study of inflammatory bowel disease (IBD) pathogenesis. Depending on the concentration, the duration and frequency of DSS administration, animals develop acute or chronic colitis or even colitis-induced dysplastic lesions. In the present work we aimed to: a) evaluate the inflammatory damage induced by different concentrations of DSS in PIE cells, and b) the capacity of immunobiotic LAB to modulate DDS-induced inflammatory immune response in PIE cells and to protect against cellular damage. PIE cells were cultured for 3 days and then challenged with different concentrations of DSS (0.1, 0.01, 0.001 and 0.0001%) for 6 hours. Cellular viability and trans epithelial resistance (TER) were studied. Our results demonstrated that DSS decrease TER and reduce viability of PIE cells in a dose dependent manner, that is in line with the general knowledge of toxic effects of DSS to epithelial cells and the increase of mucosal permeability. DSS in a concentration of 0.1% was selected for further experiments. Increased expression of TNF-α, IL-1β, IL-6, IL-16, IFN-γ, CCL2, CXCL10, and NOD2 (p