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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

Specific IgY antibody ELISA values in the egg yolk from chickens immunized with Sigma gliadin (SG), wheat gliadin (WG), and pepsin-trypsin resistant gliadin (PT-gliadin) (500 μg/ml protein) in PBS, emulsified with Freund’s incomplete adjuvant. Booster immunizations were given at 2 and 6 weeks after the initial immunization. Values are the mean of quadruple samples, with vertical bars indicating the standard deviation. Arrows indicated times of immunization
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Fig1: Specific IgY antibody ELISA values in the egg yolk from chickens immunized with Sigma gliadin (SG), wheat gliadin (WG), and pepsin-trypsin resistant gliadin (PT-gliadin) (500 μg/ml protein) in PBS, emulsified with Freund’s incomplete adjuvant. Booster immunizations were given at 2 and 6 weeks after the initial immunization. Values are the mean of quadruple samples, with vertical bars indicating the standard deviation. Arrows indicated times of immunization

Mentions: The anti-gliadin IgY antibodies obtained from chickens immunized with Sigma gliadin, wheat gliadin or PT-gliadin were weekly titrated by indirect ELISA. As shown in Fig. 1, the titre of anti-gliadin IgY was undetectable on day 0, rapidly increased (P > 0.05) from week 2 to 4, and then remained relatively constant (P > 0.05) during week 5–7 periods. Among three gliadins, chickens produced the highest anti-gliadin IgY in response to wheat gliadin (P > 0.05), throughout the immunization period. Therefore, anti-wheat gliadin IgY was used for further studies.Fig. 1


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)

Specific IgY antibody ELISA values in the egg yolk from chickens immunized with Sigma gliadin (SG), wheat gliadin (WG), and pepsin-trypsin resistant gliadin (PT-gliadin) (500 μg/ml protein) in PBS, emulsified with Freund’s incomplete adjuvant. Booster immunizations were given at 2 and 6 weeks after the initial immunization. Values are the mean of quadruple samples, with vertical bars indicating the standard deviation. Arrows indicated times of immunization
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig1: Specific IgY antibody ELISA values in the egg yolk from chickens immunized with Sigma gliadin (SG), wheat gliadin (WG), and pepsin-trypsin resistant gliadin (PT-gliadin) (500 μg/ml protein) in PBS, emulsified with Freund’s incomplete adjuvant. Booster immunizations were given at 2 and 6 weeks after the initial immunization. Values are the mean of quadruple samples, with vertical bars indicating the standard deviation. Arrows indicated times of immunization
Mentions: The anti-gliadin IgY antibodies obtained from chickens immunized with Sigma gliadin, wheat gliadin or PT-gliadin were weekly titrated by indirect ELISA. As shown in Fig. 1, the titre of anti-gliadin IgY was undetectable on day 0, rapidly increased (P > 0.05) from week 2 to 4, and then remained relatively constant (P > 0.05) during week 5–7 periods. Among three gliadins, chickens produced the highest anti-gliadin IgY in response to wheat gliadin (P > 0.05), throughout the immunization period. Therefore, anti-wheat gliadin IgY was used for further studies.Fig. 1

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