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Diabetes-specific HLA-DR-restricted proinflammatory T-cell response to wheat polypeptides in tissue transglutaminase antibody-negative patients with type 1 diabetes.

Mojibian M, Chakir H, Lefebvre DE, Crookshank JA, Sonier B, Keely E, Scott FW - Diabetes (2009)

Bottom Line: HLA-DQ2, the major celiac disease risk gene, was not significantly different.T-cell reactivity to WPs was frequently present in type 1 diabetic patients and associated with HLA-DR4 but not HLA-DQ2.The presence of an HLA-DR-restricted Th1 and Th17 response to WPs in a subset of patients indicates a diabetes-related inflammatory state in the gut immune tissues associated with defective oral tolerance and possibly gut barrier dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Chronic Disease Program, Ottawa Hospital Research Institute, Ottawa, Canada.

ABSTRACT

Objective: There is evidence of gut barrier and immune system dysfunction in some patients with type 1 diabetes, possibly linked with exposure to dietary wheat polypeptides (WP). However, questions arise regarding the frequency of abnormal immune responses to wheat and their nature, and it remains unclear whether such responses are diabetes specific.

Research design and methods: In type 1 diabetic patients and healthy control subjects, the immune response of peripheral CD3(+) T-cells to WPs, ovalbumin, gliadin, alpha-gliadin 33-mer peptide, tetanus toxoid, and phytohemagglutinin was measured using a carboxyfluorescein diacetate succinimidyl ester (CFSE) proliferation assay. T-helper cell type 1 (Th1), Th2, and Th17 cytokines were analyzed in WP-stimulated peripheral blood mononuclear cell (PBMNC) supernatants, and HLA was analyzed by PCR.

Results: Of 42 patients, 20 displayed increased CD3(+) T-cell proliferation to WPs and were classified as responders; proliferative responses to other dietary antigens were less pronounced. WP-stimulated PBMNCs from patients showed a mixed proinflammatory cytokine response with large amounts of IFN-gamma, IL-17A, and increased TNF. HLA-DQ2, the major celiac disease risk gene, was not significantly different. Nearly all responders carried the diabetes risk gene HLA-DR4. Anti-DR antibodies blocked the WP response and inhibited secretion of Th1 and Th17 cytokines. High amounts of WP-stimulated IL-6 were not blocked.

Conclusions: T-cell reactivity to WPs was frequently present in type 1 diabetic patients and associated with HLA-DR4 but not HLA-DQ2. The presence of an HLA-DR-restricted Th1 and Th17 response to WPs in a subset of patients indicates a diabetes-related inflammatory state in the gut immune tissues associated with defective oral tolerance and possibly gut barrier dysfunction.

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Role of HLA-DR in the type 1 diabetes T-cell and cytokine response to WPs. A: The T-cell response to wheat protein (6.2 μg/ml) was calculated as CDI in 14 patients and compared with cells cultured in the presence of anti–HLA-DR monoclonal antibody (5 μg/ml) or with isotype control antibody. B: Cytokine profiles of supernatants from WP-stimulated PBMNC of patients were evaluated in the absence and presence of monoclonal HLA-DR antibodies using Th1/Th2 CBAs for IFN-γ, IL-4, TNF, IL-10, and IL-6 or an ELISA kit for IL-17A.
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Figure 5: Role of HLA-DR in the type 1 diabetes T-cell and cytokine response to WPs. A: The T-cell response to wheat protein (6.2 μg/ml) was calculated as CDI in 14 patients and compared with cells cultured in the presence of anti–HLA-DR monoclonal antibody (5 μg/ml) or with isotype control antibody. B: Cytokine profiles of supernatants from WP-stimulated PBMNC of patients were evaluated in the absence and presence of monoclonal HLA-DR antibodies using Th1/Th2 CBAs for IFN-γ, IL-4, TNF, IL-10, and IL-6 or an ELISA kit for IL-17A.

Mentions: To confirm the role of HLA-DR in the T-cell response to WP, we evaluated the blocking effect of anti–HLA-DR monoclonal antibodies on PBMNC of 14 type 1 diabetic patients. Monoclonal anti-DR antibodies blocked T-cell responses to WPs in type 1 diabetic patients, whereas isotype control antibody did not prevent the T-cell response (Fig. 5A). Anti–HLA-DR4 antibody had only a small inhibitory effect on T-cell response to tetanus toxoid (supplementary Fig. S3). CBA cytokine analysis of the culture supernatants from WP-stimulated PBMNC of type 1 diabetic patients showed that monoclonal anti-DR antibody significantly inhibited secretion of Th1 cytokines but not Th2 cytokines (Fig. 5B). WP-induced production of IL-17A was also blocked by the addition of anti-DR (P = 0.008) (Fig. 5B).


Diabetes-specific HLA-DR-restricted proinflammatory T-cell response to wheat polypeptides in tissue transglutaminase antibody-negative patients with type 1 diabetes.

Mojibian M, Chakir H, Lefebvre DE, Crookshank JA, Sonier B, Keely E, Scott FW - Diabetes (2009)

Role of HLA-DR in the type 1 diabetes T-cell and cytokine response to WPs. A: The T-cell response to wheat protein (6.2 μg/ml) was calculated as CDI in 14 patients and compared with cells cultured in the presence of anti–HLA-DR monoclonal antibody (5 μg/ml) or with isotype control antibody. B: Cytokine profiles of supernatants from WP-stimulated PBMNC of patients were evaluated in the absence and presence of monoclonal HLA-DR antibodies using Th1/Th2 CBAs for IFN-γ, IL-4, TNF, IL-10, and IL-6 or an ELISA kit for IL-17A.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2712773&req=5

Figure 5: Role of HLA-DR in the type 1 diabetes T-cell and cytokine response to WPs. A: The T-cell response to wheat protein (6.2 μg/ml) was calculated as CDI in 14 patients and compared with cells cultured in the presence of anti–HLA-DR monoclonal antibody (5 μg/ml) or with isotype control antibody. B: Cytokine profiles of supernatants from WP-stimulated PBMNC of patients were evaluated in the absence and presence of monoclonal HLA-DR antibodies using Th1/Th2 CBAs for IFN-γ, IL-4, TNF, IL-10, and IL-6 or an ELISA kit for IL-17A.
Mentions: To confirm the role of HLA-DR in the T-cell response to WP, we evaluated the blocking effect of anti–HLA-DR monoclonal antibodies on PBMNC of 14 type 1 diabetic patients. Monoclonal anti-DR antibodies blocked T-cell responses to WPs in type 1 diabetic patients, whereas isotype control antibody did not prevent the T-cell response (Fig. 5A). Anti–HLA-DR4 antibody had only a small inhibitory effect on T-cell response to tetanus toxoid (supplementary Fig. S3). CBA cytokine analysis of the culture supernatants from WP-stimulated PBMNC of type 1 diabetic patients showed that monoclonal anti-DR antibody significantly inhibited secretion of Th1 cytokines but not Th2 cytokines (Fig. 5B). WP-induced production of IL-17A was also blocked by the addition of anti-DR (P = 0.008) (Fig. 5B).

Bottom Line: HLA-DQ2, the major celiac disease risk gene, was not significantly different.T-cell reactivity to WPs was frequently present in type 1 diabetic patients and associated with HLA-DR4 but not HLA-DQ2.The presence of an HLA-DR-restricted Th1 and Th17 response to WPs in a subset of patients indicates a diabetes-related inflammatory state in the gut immune tissues associated with defective oral tolerance and possibly gut barrier dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Chronic Disease Program, Ottawa Hospital Research Institute, Ottawa, Canada.

ABSTRACT

Objective: There is evidence of gut barrier and immune system dysfunction in some patients with type 1 diabetes, possibly linked with exposure to dietary wheat polypeptides (WP). However, questions arise regarding the frequency of abnormal immune responses to wheat and their nature, and it remains unclear whether such responses are diabetes specific.

Research design and methods: In type 1 diabetic patients and healthy control subjects, the immune response of peripheral CD3(+) T-cells to WPs, ovalbumin, gliadin, alpha-gliadin 33-mer peptide, tetanus toxoid, and phytohemagglutinin was measured using a carboxyfluorescein diacetate succinimidyl ester (CFSE) proliferation assay. T-helper cell type 1 (Th1), Th2, and Th17 cytokines were analyzed in WP-stimulated peripheral blood mononuclear cell (PBMNC) supernatants, and HLA was analyzed by PCR.

Results: Of 42 patients, 20 displayed increased CD3(+) T-cell proliferation to WPs and were classified as responders; proliferative responses to other dietary antigens were less pronounced. WP-stimulated PBMNCs from patients showed a mixed proinflammatory cytokine response with large amounts of IFN-gamma, IL-17A, and increased TNF. HLA-DQ2, the major celiac disease risk gene, was not significantly different. Nearly all responders carried the diabetes risk gene HLA-DR4. Anti-DR antibodies blocked the WP response and inhibited secretion of Th1 and Th17 cytokines. High amounts of WP-stimulated IL-6 were not blocked.

Conclusions: T-cell reactivity to WPs was frequently present in type 1 diabetic patients and associated with HLA-DR4 but not HLA-DQ2. The presence of an HLA-DR-restricted Th1 and Th17 response to WPs in a subset of patients indicates a diabetes-related inflammatory state in the gut immune tissues associated with defective oral tolerance and possibly gut barrier dysfunction.

Show MeSH
Related in: MedlinePlus