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

Gujral N, Suh JW, Sunwoo HH - BMC Immunol. (2015)

Bottom Line: Caco-2 monolayers were then evaluated for effects of gliadin and/or anti-wheat gliadin IgY after 24 h exposure.Among other conditions, anti-wheat gliadin IgY at a ratio of 1:3,000 (anti-gliadin IgY: PT-gliadin) significantly prevented 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).Anti-gliadin IgY, therefore has potential to be used as an oral passive antibody therapy to treat CD.

View Article: PubMed Central - PubMed

Affiliation: 3142G Katz Group Centre for Pharmacy & Health Research, Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11361 - 87 Ave, Edmonton, AB, T6G 2E1, Canada. gujral@ualberta.ca.

ABSTRACT

Background: Pepsin-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.

Methods: Caco-2 (passages 20-24) monolayers were subjected to 7 experimental conditions (n=3 each): phosphate buffered saline (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.

Results: Among other conditions, anti-wheat gliadin IgY at a ratio of 1:3,000 (anti-gliadin IgY: PT-gliadin) significantly prevented 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).

Conclusion: The 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.

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Related in: MedlinePlus

Pro-inflammatory cytokine (IL-1β and TNF-α) in Caco-2 cell. Results are expressed as mean ± SD (n = 9). * indicates statistically significant decrease in TNF-α production (P < 0.05)
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Fig5: Pro-inflammatory cytokine (IL-1β and TNF-α) in Caco-2 cell. Results are expressed as mean ± SD (n = 9). * indicates statistically significant decrease in TNF-α production (P < 0.05)

Mentions: Figure 5 illustrates that PT-gliadin stimulated the synthesis of cytokines, IL-1β and TNF-α in Caco-2 cells after 24 h incubation. Upon PT-gliadin stimulation with PT-gliadin incubation with anti-gliadin IgY at a ratio of 1:6,000 [anti-gliadin IgY (10 ng) and PT-gliadin (60 μg)], a 6.77 folds higher of TNF-α content than that of IL-1β was detected in the Caco-2 cell culture supernatant. IL-1β and TNF-α concentration in the cell supernatant were significantly decreased (P < 0.05). However, other two combinations with higher content of anti-gliadin IgY of 1: 3,000 [anti-gliadin IgY (20 ng) and PT-gliadin (60 μg)], and 1,500 [anti-gliadin IgY (40 ng) and PT-gliadin (60 μg)], showed undetectable levels of TNF-α. On the other hand, IL-1β levels remained undetectable with all three ratios of anti-gliadin IgY co-incubations with PT-gliadin. No cytokines were detected in cultures exposed to control PBS, PD-casein, and non-specific IgY.Fig. 5


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

Gujral N, Suh JW, Sunwoo HH - BMC Immunol. (2015)

Pro-inflammatory cytokine (IL-1β and TNF-α) in Caco-2 cell. Results are expressed as mean ± SD (n = 9). * indicates statistically significant decrease in TNF-α production (P < 0.05)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4495697&req=5

Fig5: Pro-inflammatory cytokine (IL-1β and TNF-α) in Caco-2 cell. Results are expressed as mean ± SD (n = 9). * indicates statistically significant decrease in TNF-α production (P < 0.05)
Mentions: Figure 5 illustrates that PT-gliadin stimulated the synthesis of cytokines, IL-1β and TNF-α in Caco-2 cells after 24 h incubation. Upon PT-gliadin stimulation with PT-gliadin incubation with anti-gliadin IgY at a ratio of 1:6,000 [anti-gliadin IgY (10 ng) and PT-gliadin (60 μg)], a 6.77 folds higher of TNF-α content than that of IL-1β was detected in the Caco-2 cell culture supernatant. IL-1β and TNF-α concentration in the cell supernatant were significantly decreased (P < 0.05). However, other two combinations with higher content of anti-gliadin IgY of 1: 3,000 [anti-gliadin IgY (20 ng) and PT-gliadin (60 μg)], and 1,500 [anti-gliadin IgY (40 ng) and PT-gliadin (60 μg)], showed undetectable levels of TNF-α. On the other hand, IL-1β levels remained undetectable with all three ratios of anti-gliadin IgY co-incubations with PT-gliadin. No cytokines were detected in cultures exposed to control PBS, PD-casein, and non-specific IgY.Fig. 5

Bottom Line: Caco-2 monolayers were then evaluated for effects of gliadin and/or anti-wheat gliadin IgY after 24 h exposure.Among other conditions, anti-wheat gliadin IgY at a ratio of 1:3,000 (anti-gliadin IgY: PT-gliadin) significantly prevented 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).Anti-gliadin IgY, therefore has potential to be used as an oral passive antibody therapy to treat CD.

View Article: PubMed Central - PubMed

Affiliation: 3142G Katz Group Centre for Pharmacy & Health Research, Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11361 - 87 Ave, Edmonton, AB, T6G 2E1, Canada. gujral@ualberta.ca.

ABSTRACT

Background: Pepsin-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.

Methods: Caco-2 (passages 20-24) monolayers were subjected to 7 experimental conditions (n=3 each): phosphate buffered saline (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.

Results: Among other conditions, anti-wheat gliadin IgY at a ratio of 1:3,000 (anti-gliadin IgY: PT-gliadin) significantly prevented 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).

Conclusion: The 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.

Show MeSH
Related in: MedlinePlus