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

RENATA CURCIARELLO; KARINA CANZIANI; MARÍA DE LOS ANGELESSERRADELL; AGUSTINA ERREA; MARTÍN RUMBO; AYELÉN HUGO; ANDRÉS ROCCA; SANTIAGO BRAYER ; ALICIA SAMBUELLI; MARTÍN YANTORNO GUSTAVO CORREA; NÉSTOR CHOPITA; GUILLERMO H. DOCENA; CECILIA MUGLIA

Washington

Congreso; International Congress of Mucosal Immunology; 2017

Society for Mucosal Immunology

Lamina propria T cells (LPTC) are key cells in inflammatory Bowel Disease (IBD) pathogenesis, contributing to mucosal inflammation by secreting pro-inflammatory cytokines and being resistant to apoptosis. Kefir is a fermented milk with health-promoting properties.Here we have isolated and established primary cultures of LPTC from IBD patients and modulated their pro-inflammatory response using microorganisms from kefir.Colonic biopsies from IBD patients (N=12, 5 CD and 7 CU) were washed in HBSS medium with EDTA and DTT, and digested by collagenase and DNAse treatment. In order to enrich pathogen-specific T lymphocytes, 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 LPTC stimulated with anti-CD3/CD28 and probiotic bacteria, conditioned media or 10 µM lactate.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 (P˂0.05) while IL-6 and IL-8 diminished with E. durans. 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 results could contribute to future therapies for IBD.