Gene expression assessed by genome wide hybridization bead array in T84 polarized tight monolayers after challenge with celiac disease-associated bacteria and gluten [A grav, Bmix Bmix glut]

Analysis of the influence of celiac disease-associated bacteria and gluten on intestinal epithelial cells Stimulation with CD-associated bacteria and gluten was done from the apical side of polarized tight monolayers of T84 cells by replacing the medium in the upper chamber with 0.5 mL challenge medium containing 5x10^7 bacteria with or without 1 mg trypsin-treated gluten/mL. For each stimulant three parallel wells were used to carry out the experiment and wells treated with challenge medium only served as experimental controls. The challenge was done under anaerobic condition and at 37°C for 4 hours. The bacteria mix includes Prevotella jejuni (three isolates) + P. melaninogenica + P. histicola + Lachnoanaerobaculum umeaense + Actinomyces graevenitzii in ratio 1:1:1:1:1:1:1. Gluten = tryptic digest of wheat gluten.