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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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Related in: MedlinePlus

Graphic representation of the relationship between wheat-induced T-cell response and high-resolution HLA diabetes risk alleles. Box and Whisker plots with individual values for diabetic subjects. Each plot includes the mean (bold line), median (solid thin line), distribution, and range. A: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DRB1*03 and HLA-DRB1*04 or in those heterozygous for HLA-DRB1*03/*04. A CDI value greater than mean + 3 SD of the control group (CDI >14.6, dashed line) was used to define a positive proliferation response. One of the type 1 diabetic patients with HLA-DRB1*03+/*04− showed positive responses to WP (○). Type 1 diabetic patients who carried one risk allele HLA-DRB1*04 (10 of 20) or patients who carried both risk genes HLA-DRB1*03/*04 (9 of 12) showed positive reactivity to WP. High-resolution results of HLA-DR are labeled beside each individual. B: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DQB1*02 and HLA-DQB1*03 or in those heterozygous for HLA-DQB1*02/*03. High-resolution analyses for HLA-DQB1*02 and *03 are labeled beside each symbol. ↓ (beside 0201) means all individuals in the column carry HLA-DQB1*0201. (Three of the 42 type 1 diabetic individuals are neither DR3 nor DR4, and therefore they are not included in this figure. One of the seven individuals with DR3/non-DR4 in the left panel is heterozygous for DQ*02/*03 and was therefore placed in the middle column in the right panel [DQ*02*03]. Differences in high-resolution haplotypes between responders and nonresponders were not evaluated statistically because of the small sample size.)
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Figure 4: Graphic representation of the relationship between wheat-induced T-cell response and high-resolution HLA diabetes risk alleles. Box and Whisker plots with individual values for diabetic subjects. Each plot includes the mean (bold line), median (solid thin line), distribution, and range. A: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DRB1*03 and HLA-DRB1*04 or in those heterozygous for HLA-DRB1*03/*04. A CDI value greater than mean + 3 SD of the control group (CDI >14.6, dashed line) was used to define a positive proliferation response. One of the type 1 diabetic patients with HLA-DRB1*03+/*04− showed positive responses to WP (○). Type 1 diabetic patients who carried one risk allele HLA-DRB1*04 (10 of 20) or patients who carried both risk genes HLA-DRB1*03/*04 (9 of 12) showed positive reactivity to WP. High-resolution results of HLA-DR are labeled beside each individual. B: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DQB1*02 and HLA-DQB1*03 or in those heterozygous for HLA-DQB1*02/*03. High-resolution analyses for HLA-DQB1*02 and *03 are labeled beside each symbol. ↓ (beside 0201) means all individuals in the column carry HLA-DQB1*0201. (Three of the 42 type 1 diabetic individuals are neither DR3 nor DR4, and therefore they are not included in this figure. One of the seven individuals with DR3/non-DR4 in the left panel is heterozygous for DQ*02/*03 and was therefore placed in the middle column in the right panel [DQ*02*03]. Differences in high-resolution haplotypes between responders and nonresponders were not evaluated statistically because of the small sample size.)

Mentions: We analyzed HLA-DR and -DQ haplotypes in patients with type 1 diabetes and control subjects. The frequency of HLA haplotypes in the control group was similar to previously published reference populations (34) (data not shown). HLA-DRB1*03 and -DRB1*04, which are associated with type 1 diabetes, were more frequent in type 1 diabetic patients compared with control subjects. Heterozygous DR3/DR4 individuals represented 29% of our type 1 diabetic patients, but none of our control subjects was heterozygous for HLA-DR3/DR4 (data not shown). In addition, we compared the frequency of risk alleles in type 1 diabetic patients who were either responders or nonresponders to WP (Table 2). Type 1 diabetic responders to WP carried HLA-DRB1*04 in 95% of cases, which was significantly higher than in the nonresponders (59%; P = 0.01) (Table 2). Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DRB1*03 or HLA-DRB1*04 or in those heterozygous for HLA-DRB1*03/*04 is shown in Fig. 4. One of seven type 1 diabetic patients with HLA-DRB1*03+/*04− showed positive responses to WP. In contrast, 10 of 20 type 1 diabetic patients who carried one risk allele HLA-DRB1*04 or 9 of 12 patients who carried both risk genes HLA-DRB1*03/*04 showed positive reactivity to WP.


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)

Graphic representation of the relationship between wheat-induced T-cell response and high-resolution HLA diabetes risk alleles. Box and Whisker plots with individual values for diabetic subjects. Each plot includes the mean (bold line), median (solid thin line), distribution, and range. A: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DRB1*03 and HLA-DRB1*04 or in those heterozygous for HLA-DRB1*03/*04. A CDI value greater than mean + 3 SD of the control group (CDI >14.6, dashed line) was used to define a positive proliferation response. One of the type 1 diabetic patients with HLA-DRB1*03+/*04− showed positive responses to WP (○). Type 1 diabetic patients who carried one risk allele HLA-DRB1*04 (10 of 20) or patients who carried both risk genes HLA-DRB1*03/*04 (9 of 12) showed positive reactivity to WP. High-resolution results of HLA-DR are labeled beside each individual. B: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DQB1*02 and HLA-DQB1*03 or in those heterozygous for HLA-DQB1*02/*03. High-resolution analyses for HLA-DQB1*02 and *03 are labeled beside each symbol. ↓ (beside 0201) means all individuals in the column carry HLA-DQB1*0201. (Three of the 42 type 1 diabetic individuals are neither DR3 nor DR4, and therefore they are not included in this figure. One of the seven individuals with DR3/non-DR4 in the left panel is heterozygous for DQ*02/*03 and was therefore placed in the middle column in the right panel [DQ*02*03]. Differences in high-resolution haplotypes between responders and nonresponders were not evaluated statistically because of the small sample size.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 4: Graphic representation of the relationship between wheat-induced T-cell response and high-resolution HLA diabetes risk alleles. Box and Whisker plots with individual values for diabetic subjects. Each plot includes the mean (bold line), median (solid thin line), distribution, and range. A: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DRB1*03 and HLA-DRB1*04 or in those heterozygous for HLA-DRB1*03/*04. A CDI value greater than mean + 3 SD of the control group (CDI >14.6, dashed line) was used to define a positive proliferation response. One of the type 1 diabetic patients with HLA-DRB1*03+/*04− showed positive responses to WP (○). Type 1 diabetic patients who carried one risk allele HLA-DRB1*04 (10 of 20) or patients who carried both risk genes HLA-DRB1*03/*04 (9 of 12) showed positive reactivity to WP. High-resolution results of HLA-DR are labeled beside each individual. B: Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DQB1*02 and HLA-DQB1*03 or in those heterozygous for HLA-DQB1*02/*03. High-resolution analyses for HLA-DQB1*02 and *03 are labeled beside each symbol. ↓ (beside 0201) means all individuals in the column carry HLA-DQB1*0201. (Three of the 42 type 1 diabetic individuals are neither DR3 nor DR4, and therefore they are not included in this figure. One of the seven individuals with DR3/non-DR4 in the left panel is heterozygous for DQ*02/*03 and was therefore placed in the middle column in the right panel [DQ*02*03]. Differences in high-resolution haplotypes between responders and nonresponders were not evaluated statistically because of the small sample size.)
Mentions: We analyzed HLA-DR and -DQ haplotypes in patients with type 1 diabetes and control subjects. The frequency of HLA haplotypes in the control group was similar to previously published reference populations (34) (data not shown). HLA-DRB1*03 and -DRB1*04, which are associated with type 1 diabetes, were more frequent in type 1 diabetic patients compared with control subjects. Heterozygous DR3/DR4 individuals represented 29% of our type 1 diabetic patients, but none of our control subjects was heterozygous for HLA-DR3/DR4 (data not shown). In addition, we compared the frequency of risk alleles in type 1 diabetic patients who were either responders or nonresponders to WP (Table 2). Type 1 diabetic responders to WP carried HLA-DRB1*04 in 95% of cases, which was significantly higher than in the nonresponders (59%; P = 0.01) (Table 2). Distribution of T-cell proliferation (CDI) to WP in type 1 diabetic patients with HLA-DRB1*03 or HLA-DRB1*04 or in those heterozygous for HLA-DRB1*03/*04 is shown in Fig. 4. One of seven type 1 diabetic patients with HLA-DRB1*03+/*04− showed positive responses to WP. In contrast, 10 of 20 type 1 diabetic patients who carried one risk allele HLA-DRB1*04 or 9 of 12 patients who carried both risk genes HLA-DRB1*03/*04 showed positive reactivity to WP.

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