Isolation and Modulation of Pathogen specific T cells from inflammatory Bowel Disease Patients

CANZIANI, K.; HUGO, A; SAMBUELLI, A.; CHOPITA, N; CURCIARELLO, R; RUMBO, M; BRAYER, S; CORREA G; SERRADELL, M; MUGLIA, C. I; ROCCA, A; YANTORNO, M; DOCENA, G.

Mar del Plata

Congreso; SAIC.SAI.SAFE 2016; 2016

Lamina propria T cells (LPTC) are key cells in inflammatory Bowel Disease (IBD) pathogenesis, contributing to mucosal inflammation by pro-inflammatory cytokines secretion and apoptosis resistance. Kefir is a fermented milk with health-promoting properties.We have isolated and established primary cultures of LPTC from IBD patients and modulated their pro-inflammatory response using microorganisms from kefir.Colonic biopsies or colonic mucosa from surgical specimens from IBD patients (N=12, 5 CD and 7 CU) were washed in HBSS buffer with EDTA and DTT, and digested with collagenase and 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 (Lactobacillus kefiri and Enterococcus durans) were evaluated for their modulation capacity on LPTC, by proliferation assays (CFSE) and cytokine secretion (TNF, IL-6, IL-8 and IL-10 by ELISA) of stimulated LPTC (anti-CD3/anti-CD28 and probiotic bacteria, conditioned media or 10 µM lactate stimuli).LPTC lines specific for EA E. coli were developed for all patients. Cell proliferation of activated lymphocytes decreased with L. kefiri and E. durans (proliferation index: 3.0±0.5 vs 0.9±0.3 and 0.56±0.3 respectively; unstimulated control: 1.0±0.1). TNF, IL-6 and IL-8 secretion was decreased in activated LPTC incubated with L. kefiri compared to medium while IL-6 and IL-8 diminished with E. durans (P˂0.05). Intermediate results 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 for IBD.