<i>Vibrio cholerae</i> toxin coregulated pilus provokes inflammatory responses in Coculture model of Caco‐2 and peripheral blood mononuclear cells (PBMC) leading to increased colonization

The aim of this study was to assess the modulatory effect TcpA in expression CEACAM1 adhesin molecule and IL-1, IL-8, TNF-α pro-inflammatory cytokines Coculture model Caco-2/PBMC (peripheral blood mononuclear cell) that can mimic intestinal milieu. gene from Vibrio cholerae ATCC14035 cloned pET-28a transformed into Escherichia coli Bl-21. recombinant TcpA-His6 protein expressed purified using Ni-column chromatography. sequencing plasmid Western blot analysis confirmed identity rTcp. cytotoxicity different concentrations for human colon carcinoma cell line (human colorectal adenocarcinoma [Caco-2 cell]) assessed by MTT assay showed viability 92%, 82%, 70%, 10 µg/mL after 24, 48, 72 h, respectively. Co-cultures Caco-2 PBMCs were used milieu treated with rTcpA (1, 5, 10, 50 µg/mL). Our data about 2.04-, 3.37-, 3.68-, 42.7-fold increase expression, respectively, compared nontreated Coculture. Moreover, genes significantly increased up 15.75-, 7.04-, 80.95-folds, In conclusion, V. induces statistically significant dose-dependent stimulatory on TNF-α, IL-,1, IL-8 expression. Of these, much more affected which, consequently, elevated level IECs. This suggests is a critical effector as an inducer adhesion potential cholera well inflammatory responses host tissue.