Identification of cross-reactive B-cell epitopes between Gly m 5.0101 [Glycine max] and α-casein [Bos Taurus] by epitope mapping - mass spectrometry.

CURCIARELLO RENATA; RUMBO MARTÍN; BRAYER SANTIAGO; CORREA GUSTAVO; DOCENA GUILLERMO; SERRADELL MARÍA; ROCCA ANDRES; YANTORNO MARTIN; MUGLIA CECILIA; CANZIANI KARINA; HUGO AYELEN; SAMBUELLI ALICIA; CHOPITA NÉSTOR

Mar del Plata

Congreso; Congreso conjunto SAIC-SAI-SAFE, LXIV Reunión Anual de SAI; 2016

SAI

Inflammatory bowel diseases (IBD) are a complex group of pathologies amongst which ulcerative colitis (UC) andCrohn´s disease (CD) are the most conspicuous. Lamina propria T cells (LPTC) are key cells in IBD pathogenesis,contributing to mucosal inflammation by secreting pro-inflammatory cytokines. It has been shown that LPTC in IBD areresistant to apoptosis. Kefir is a fermented milk that has been used in Eastern Europe with health-promoting properties.The aim of this work was to isolate and establish primary cultures of LPTC from IBD patients and modulate their proinflammatoryresponse using microorganisms from kefir.Colonic biopsies from IBD patients (N=8, 5 CD and 3 UC) were washed in HBSS medium with EDTA and DTT, anddigested by collagenase and DNAse treatment. In order to select pathogen-specific T lymphocytes, cells were culturedwith extracts of enteroadhesive (EA) Escherichia coli and IL-2. Microorganisms from kefir (Lactobacilluskefiri and Enterococcus durans) were evaluated for their modulation capacity on LPTC by proliferation assays (flowcytometry with CFSE) and cytokine secretion (TNF and IL-10 by ELISA) of T cell lines stimulated with anti-CD3 andanti-CD28 followed by culture with probiotic bacteria.LPTC lines specific for EA E. coli were developed for all patients. Cell proliferation of activated lymphocytes decreasedwith the presence of L. kefiri and E. durans (proliferation index: 3.0±0.54 vs 1.3±0.33 and 0.56±0.28 respectively;unstimulated control: 1.02±0.03). Besides, TNF secretion was dampened in activated LPTC incubated with L.kefiri compared to medium (P˂0.05). No significant differences were observed for IL-10 secretion.Our results show that these probiotics from kefir modulate pathogen specific activated T cells form IBD patients. Thesepromising results could provide the basis for future therapies in IBD patients.