Effect of anti-gliadin IgY antibody on epithelial intestinal integrity and inflammatory response induced by gliadin

Abstract

BackgroundPepsin-trypsin resistant gliadin (PT-gliadin) promotes intestinal tissue inflammation and increases paracellular permeability of immunogenic gliadin peptides into the lamina propria. This leads to the complications seen in the pathogenesis of celiac disease (CD). In this study, specific anti-gliadin IgY antibody was produced and evaluated for its efficacy on gliadin induced intestinal integrity impairment and proinflammatory effects on intestinal epithelial (Caco-2) cell culture model for CD.MethodsCaco-2 (passages 20-24) monolayers were subjected to 7 experimental conditions (n=3 each): phosphatebufferedsaline (PBS; control), pancreatic digested-casein (PD-casein; negative control), PT-gliadin (positive control), non-specific IgY with PT-gliadin, and anti-wheat gliadin IgY with PT-gliadin at a ratio of 1:6,000, 1:3,000 and 1:1,500. Caco-2 monolayers were then evaluated for effects of gliadin and/or anti-wheat gliadin IgY after 24 h exposure. Enzyme-linked immunosorbent assay (ELISA) was used to quantify anti-inflammatory markers (TNF-α and IL-1β) 5 days after cells were exposed to PT-gliadin and/or anti-wheat gliadin IgY.ResultsAmong other conditions, anti-wheat gliadin IgY at a ratio of 1:3,000 (anti-gliadin IgY: PT-gliadin) significantlyprevented gliadin toxicity on Caco-2 by maintaining intestinal integrity, inhibiting phenol red permeation, and inhibiting gliadin absorption and production of proinflammatory cytokines (TNF-α and IL-1β) as compared to PT-gliadin stimulated cultures (P < 0.05).ConclusionThe anti-wheat gliadin IgY antibody produced in this study has proved to inhibit absorption of gliadin and gliadin-induced inflammatory response in Caco2 cell culture model of CD. Anti-gliadin IgY, therefore has potential to be used as an oral passive antibody therapy to treat CD.

Exploring Novel Cocrystalline Forms of Oxyresveratrol to Enhance Aqueous Solubility and Permeability across a Cell Monolayer

“…34,35) The TEER measurement and phenol red assays are widely used in the evaluation of the monolayer integrity. [36][37][38][39] since they are simple and nonradioactive. In this study, we therefore used the TEER measurement and phenol red assay to evaluate the monolayer integrity before and during the cellular transport experiment.…”

Current and novel therapeutic strategies in celiac disease

“…These antibodies were able to bind with gliadin preparations in simulated gastric and intestinal fluids in in vitro models and were also able to decrease gliadin absorption to <1% from the gastrointestinal tract of mice. In another human tissue in vitro study, anti-gliadin IgY antibodies in combination with pepsin trypsin digested gliadin extracts demonstrated preservation of intestinal integrity and decreased production of pro-inflammatory cytokines (TNF-α and IL-1β) on Caco-2 membranes compared to pepsin trypsin digests alone (55). A phase 1/2 open label single group trial using 1000 mg of IgY with each meal in celiac patients with mild to moderate symptoms on a GFD will assess safety and efficacy of this drug in humans.…”

Sangiovese cv Pomace Seeds Extract-Fortified Kefir Exerts Anti-Inflammatory Activity in an In Vitro Model of Intestinal Epithelium Using Caco-2 Cells

“…After 60 min of incubation at 37 • C, 100 µL was collected from the basolateral chamber and added to 10 µL of 1N NaOH in 96 multiwell plates. Phenol red leakage was measured using a SAFAS MP96 spectrophotometer (Safas, Principality of Monaco) by recording the absorbance at 540 nm, and quantified according to a calibration curve obtained with serial dilutions (500-0.5 mM) of phenol red solution in HBSS [43][44][45].…”