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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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T-cell proliferation in response to WPs and other antigens or mitogen. 1.2 × 106 CFSE-labeled PBMNCs from patients with type 1 diabetes (T1D) or healthy control subjects were cultured for 8 days in the absence or presence of WP, ovalbumin (OVA), gliadin, 33-mer, insulin, tetanus toxoid (TT), or phytohemagglutinin (PHA) (see research design and methods for details). On day 8, cells were stained with Cy-chrome–conjugated anti-CD3 monoclonal antibody. CDI was calculated. A CDI value greater than the control mean + 3 SD (CDI >14.6) was used to define a positive response to WP. P values indicate statistical difference compared with control subjects. The horizontal lines indicate the means.
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Figure 2: T-cell proliferation in response to WPs and other antigens or mitogen. 1.2 × 106 CFSE-labeled PBMNCs from patients with type 1 diabetes (T1D) or healthy control subjects were cultured for 8 days in the absence or presence of WP, ovalbumin (OVA), gliadin, 33-mer, insulin, tetanus toxoid (TT), or phytohemagglutinin (PHA) (see research design and methods for details). On day 8, cells were stained with Cy-chrome–conjugated anti-CD3 monoclonal antibody. CDI was calculated. A CDI value greater than the control mean + 3 SD (CDI >14.6) was used to define a positive response to WP. P values indicate statistical difference compared with control subjects. The horizontal lines indicate the means.

Mentions: Some patients displayed increased T-cell responses to other dietary antigens including ovalbumin, an irrelevant dietary antigen, and to the celiac-related antigens (wheat gliadin and α-gliadin 33-mer peptide) as well as the type 1 diabetes autoantigen insulin (P = 0.02, P = 0.03, P = 0.001, and P = 0.03, respectively) (Fig. 2). We did not find significant differences between type 1 diabetic patients and control subjects in response to the recall antigen, tetanus toxoid, or the T-cell mitogen phytohemagglutinin (P = 0.2) (Fig. 2). There was a positive correlation between WP T-cell responses and T-cell responses to ovalbumin, gliadin, and α-gliadin 33-mer peptide in type 1 diabetic patients (P = 0.01, P = 0.002, and P = 0.0001; data not shown). In contrast, we detected a weak, nonsignificant correlation between WP T-cell response and insulin T-cell response in type 1 diabetic patients (P = 0.06; data not shown).


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)

T-cell proliferation in response to WPs and other antigens or mitogen. 1.2 × 106 CFSE-labeled PBMNCs from patients with type 1 diabetes (T1D) or healthy control subjects were cultured for 8 days in the absence or presence of WP, ovalbumin (OVA), gliadin, 33-mer, insulin, tetanus toxoid (TT), or phytohemagglutinin (PHA) (see research design and methods for details). On day 8, cells were stained with Cy-chrome–conjugated anti-CD3 monoclonal antibody. CDI was calculated. A CDI value greater than the control mean + 3 SD (CDI >14.6) was used to define a positive response to WP. P values indicate statistical difference compared with control subjects. The horizontal lines indicate the means.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 2: T-cell proliferation in response to WPs and other antigens or mitogen. 1.2 × 106 CFSE-labeled PBMNCs from patients with type 1 diabetes (T1D) or healthy control subjects were cultured for 8 days in the absence or presence of WP, ovalbumin (OVA), gliadin, 33-mer, insulin, tetanus toxoid (TT), or phytohemagglutinin (PHA) (see research design and methods for details). On day 8, cells were stained with Cy-chrome–conjugated anti-CD3 monoclonal antibody. CDI was calculated. A CDI value greater than the control mean + 3 SD (CDI >14.6) was used to define a positive response to WP. P values indicate statistical difference compared with control subjects. The horizontal lines indicate the means.
Mentions: Some patients displayed increased T-cell responses to other dietary antigens including ovalbumin, an irrelevant dietary antigen, and to the celiac-related antigens (wheat gliadin and α-gliadin 33-mer peptide) as well as the type 1 diabetes autoantigen insulin (P = 0.02, P = 0.03, P = 0.001, and P = 0.03, respectively) (Fig. 2). We did not find significant differences between type 1 diabetic patients and control subjects in response to the recall antigen, tetanus toxoid, or the T-cell mitogen phytohemagglutinin (P = 0.2) (Fig. 2). There was a positive correlation between WP T-cell responses and T-cell responses to ovalbumin, gliadin, and α-gliadin 33-mer peptide in type 1 diabetic patients (P = 0.01, P = 0.002, and P = 0.0001; data not shown). In contrast, we detected a weak, nonsignificant correlation between WP T-cell response and insulin T-cell response in type 1 diabetic patients (P = 0.06; data not shown).

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