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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

IgY purification by Sephacryl S-300 chromatography. Each value of absorbance was determined by an indirect ELISA. Flow rate: 3 ml/h, Column: 1.0 × 110 cm
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Fig2: IgY purification by Sephacryl S-300 chromatography. Each value of absorbance was determined by an indirect ELISA. Flow rate: 3 ml/h, Column: 1.0 × 110 cm

Mentions: After 5–7 weeks of immunization, the egg yolks from wheat gliadin immunized chickens were pooled for purification of IgY by Sephacryl S-300 gel chromatography. The elution profile shows that fractions corresponding to Kav 0.2–0.24 contain IgY determined by indirect ELISA (Fig. 2). Yield of total IgY as shown in Table 1 was similar regardless of the different gliadin immunizations (P > 0.05). However, the concentration of specific anti-gliadin IgY was significantly higher in Sigma gliadin, wheat gliadin, and PT-gliadin immunized chickens than in the non-immunized chickens (P < 0.05). Sephacryl S-300 purified IgY fractions against Sigma gliadin, wheat gliadin, and PT-gliadin contain approximately 7.9 %, 8.1 %, and 7.7 % of specific anti-gliadin IgY in a total of IgY antibodies, respectively.Fig. 2


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)

IgY purification by Sephacryl S-300 chromatography. Each value of absorbance was determined by an indirect ELISA. Flow rate: 3 ml/h, Column: 1.0 × 110 cm
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig2: IgY purification by Sephacryl S-300 chromatography. Each value of absorbance was determined by an indirect ELISA. Flow rate: 3 ml/h, Column: 1.0 × 110 cm
Mentions: After 5–7 weeks of immunization, the egg yolks from wheat gliadin immunized chickens were pooled for purification of IgY by Sephacryl S-300 gel chromatography. The elution profile shows that fractions corresponding to Kav 0.2–0.24 contain IgY determined by indirect ELISA (Fig. 2). Yield of total IgY as shown in Table 1 was similar regardless of the different gliadin immunizations (P > 0.05). However, the concentration of specific anti-gliadin IgY was significantly higher in Sigma gliadin, wheat gliadin, and PT-gliadin immunized chickens than in the non-immunized chickens (P < 0.05). Sephacryl S-300 purified IgY fractions against Sigma gliadin, wheat gliadin, and PT-gliadin contain approximately 7.9 %, 8.1 %, and 7.7 % of specific anti-gliadin IgY in a total of IgY antibodies, respectively.Fig. 2

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