Isolation and Modulation of Pathogen-Specific T cells from Patients with Inflammatory Bowel Diseases

MUGLIA CECILIA; CURCIARELLO RENATA; CANZIANI KARINA; SERRADELL MARÍA DE LOS ANGELES; ERREA, A; RUMBO, M; AYELÉN HUGO; ANDRÉS ROCCA; SANTIAGO BRAYER; ALICIA SAMBUELLI; YANTORNO M; GUSTAVO J. CORREA; NÉSTOR CHOPITA; DOCENA GUILLERMO H.

Washington DC

Congreso; International Congress of Mucosal Immunology 2017; 2017

Society of Mucosal Immunology

Lamina propria T cells (LPTC) are key cells ininflammatory Bowel Disease (IBD) pathogenesis,contributing to mucosal inflammation by proinflammatorycytokines secretion and apoptosisresistance. Kefir is a fermented milk with healthpromotingproperties. We have isolated andestablished primary cultures of LPTC from IBD patientsand modulated their pro-inflammatory response usingmicroorganisms from kefir. Colonic biopsies or colonicmucosa from surgical specimens from IBD patients(N=12, 5 CD and 7 CU) were washed in HBSS bufferwith EDTA and DTT, and digested with collagenaseand DNAse. In order to enrich pathogen-specific LPTC,cells were cultured with extracts of enteroadhesive(EA) Escherichia coli and IL-2 for 10 days.Microorganisms from kefir (Lactobacilluskefiri and Enterococcus durans) were evaluated fortheir modulation capacity on LPTC, by proliferationassays (CFSE) and cytokine secretion (TNF, IL-6, IL-8and IL-10 by ELISA) of stimulated LPTC (anti-CD3/anti-CD28 and probiotic bacteria, conditioned media or 10μM lactate stimuli). LPTC lines specific for EA E.coli were developed for all patients. Cell proliferationof activated lymphocytes decreased with L.kefiri and E. durans (proliferation index: 3.0±0.5 vs0.9±0.3 and 0.56±0.3 respectively; unstimulatedcontrol: 1.0±0.1). TNF, IL-6 and IL-8 secretion wasdecreased in activated LPTC incubated with L.kefiri compared to medium while IL-6 and IL-8diminished with E. duransresults were found for lactate and conditioned media.No significant differences were observed for IL-10.Our results show that probiotic strains modulate pathogen specific activated T cells from IBD patients.These findings could contribute to future therapies forIBD.