Modulatory effect of Vibrio cholerae toxin co-regulated pilus on mucins, toll-like receptors and NOD genes expression in co-culture model of Caco-2 and peripheral blood mononuclear cells (PBMC)

Copyright © 2020 Elsevier Ltd. All rights reserved..

Vibrio cholerae, the causative agent of cholera, tend to colonize the small intestine as a Gram-negative pathogen. The intestinal mucus layer forms mucin physical barrier, consisted of high molecular weight proteins. Regarding the role of toxin-coregulated pilus (TCP) as one of the most important colonization factors of V. cholerae, this experimental study was designed to determine the role of TcpA in induction of mucin production and its regulatory effect on innate immunity molecules including toll like receptors (TLRs) and Nucleotide-binding oligomerization domain-containing proteins (NODs) using Caco2- PBMC co-cultures as an interactive model. The rTcpA protein of V. cholerae was expressed in BL21 Escherichia coli, purified using Ni-column chromatography and confirmed by western blotting. Nontoxic doses of rTcpA was determined on Caco-2 cell lines and different concentrations of rTcpA (1, 5, 10 and 50 μg/mL) showed a statistically significant effect on the expression of muc genes (MUC3 and MUC4) in a dose-dependent manner. This finding is supposed to facilitate physical adhesion and colonization of V. cholerae in intestinal lumen. The rTcpA moderately stimulated the expression of tlr4 and overexpressed tlr1, both of which are supposed to induce a mucosal protective response against bacterial infection. NOD2 was significantly increased which suggests that it may contribute in pro-inflammatory responses observed in cholera disease. No change in NOD1 expression was seen which might be attributed to the non-invasive nature of V. cholerae as an intestinal pathogen. In conclusion, the rTcpA protein of V. cholerae showed a statistically significant modulatory effect on the human gut epithelium gene expression which would help promising protection in prophylaxis applications.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:149

Enthalten in:

Microbial pathogenesis - 149(2020) vom: 01. Dez., Seite 104566

Sprache:

Englisch

Beteiligte Personen:

Ghasemi, Maryam [VerfasserIn]
Bakhshi, Bita [VerfasserIn]
Khashei, Reza [VerfasserIn]
Soudi, Sara [VerfasserIn]

Links:

Volltext

Themen:

9012-63-9
Cholera Toxin
Journal Article
Mucins
NLRs
TLRs
TcpA
Toll-Like Receptors
V. cholerae

Anmerkungen:

Date Completed 17.06.2021

Date Revised 17.06.2021

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1016/j.micpath.2020.104566

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM316285943