Primary human polarized small intestinal epithelial barriers respond differently to a hazardous and an innocuous protein

Abstract

An experimental platform employing human derived intestinal epithelial cell (IEC) line monolayers grown on permeable Transwell filters was previously investigated to differentiate between hazardous and innocuous proteins. This approach was effective at distinguishing these types of proteins and perturbation of monolayer integrity, particularly transepithelial electrical resistance (TEER), was the most sensitive indicator. In the current report, in vitro indicators of monolayer integrity, cytotoxicity, and inflammation were evaluated using primary (non-transformed) human polarized small intestinal epithelial barriers cultured on Transwell filters to compare effects of a hazardous protein (Clostridium difficile Toxin A [ToxA]) and an innocuous protein (bovine serum albumin [BSA]). ToxA exerted a reproducible decrease on barrier integrity at doses comparable to those producing effects observed from cell line-derived IEC monolayers, with TEER being the most sensitive indicator. In contrast, BSA, tested at concentrations substantially higher than ToxA, did not cause changes in any of the tested variables. These results demonstrate a similarity in response to certain proteins between cell line-derived polarized IEC models and a primary human polarized small intestinal epithelial barrier model, thereby reinforcing the potential usefulness of cell line-derived polarized IECs as a valid experimental platform to differentiate between hazardous and non-hazardous proteins.

Human Primary Cell-Based Organotypic Microtissues for Modeling Small Intestinal Drug Absorption

“…Nonetheless, additional studies utilizing a larger set of model drugs from different chemical classes and interlaboratory validation studies are needed to further establish the utility of the intestinal microtissue model for drug permeation studies. In addition to predicting drug absorption, we anticipate that the reconstructed SMI microtissues will have utility in studies aimed at elucidating: a) drug-induced gastrointestinal (GI) toxicity (43, 44) under acute and extended exposure conditions, b) GI inflammation, c) innate immune responses, d) GI tract infection (45), e) effects of probiotics (46), and f) disease modeling.…”

Lactobacillus brevis 23017 Relieves Mercury Toxicity in the Colon by Modulation of Oxidative Stress and Inflammation Through the Interplay of MAPK and NF-κB Signaling Cascades

“…Moreover, recent research has revealed that oral Hg exposure affects the gut ecology, increasing inflammation and susceptibility to colitis in mice (Stejskal, 2013 ). Research has shown that Hg exposure causes serious intestinal disease, including damage to tight junction proteins and necrosis of epithelial cells through an inflammatory response (Toomey et al, 2014 ; Eaton et al, 2017 ). Disruption of the intestinal barrier has been related to gut immunity and oxidative stress induced by Hg exposure (Laporte et al, 2002 ; Hoyle and Handy, 2005 ).…”

Scientific Opinion on development needs for the allergenicity and protein safety assessment of food and feed products derived from biotechnology

“…For the risk assessment, it is necessary to include information on the source of the gene/protein (history of use), the amino acid sequence for performing similarity searches, and on structural properties such as susceptibility to enzymatic degradation. For the latter, although the isolation or purification of the newly expressed protein is needed, it might not be possible or practical because of the presence of a large number of proteins or technical difficulties of intractable proteins (Bushey et al, 2014 ; Eaton et al, 2017 ). Synthetic biology‐derived plants (and their derived food and feed products) may arrive on the market in the near future with an increased level of complexity compared to conventional GM plants (e.g.…”