The highest genetic type 1 diabetes risk is conferred by HLA class II haplotypes defined by alleles at the HLA-DR and -DQ loci. The combination of HLA-DQA1*03:01 and DQB1*03:02 alleles (summarized as ‘HLA-DQ8′) is reported to be among the two most prevalent HLA class II haplotypes in Caucasian type 1 diabetes patients. This classification is based on conventional genotyping of exon 2 of the DQ gene locus and excludes exon 3. In this study, HLA genotyping on the type 1 diabetes susceptibility loci HLA-DRB1, DQA1 and DQB1 was performed using a high-resolution next generation sequencing method. In addition to the routinely examined exon 2, exon 3 was also sequenced. Samples from 229 children with type 1 diabetes were included and compared to a cohort of 9,786 controls. In addition to previously described HLA-DQ haplotypes in type 1 diabetes patients, we found that as well as HLA-DQA1*03:01,HLA-DQA1*03:03 also contributed to HLA-DQ8. HLA-DQA1*03:03 differs from HLA-DQA1*03:01 by one nucleotide substitution in exon 3 at position 160, leading to a single amino acid replacement. DRB1*04:05 was exclusively associated with DQA1*03:03 whereas the DRB1*04:01 haplotype comprised either DQA1*03:01 or DQA1*03:03. Significantly increased type 1 diabetes risk was confirmed for all these haplotypes with only minor differences between DQA1*03:01 and DQA1*03:03 alleles. This study identified the HLA-DQA1*03:03 allele as an addition to the already known type 1 diabetes risk haplotypes, and can contribute to more precise HLA genotyping approaches.

The important role of IL‐7 in the generation of self‐reactive T‐cells in autoimmune diseases is well established. Recent studies on autoimmunity‐associated genetic polymorphisms indicated that differential IL‐7 receptor (IL‐7R) expression of monocytes may play a role in the underlying pathogenesis. The relevance of IL‐7‐mediated monocyte functions in type 1 diabetes remains elusive. In the present study, we characterized monocyte phenotype and IL‐7‐mediated effects in children with type 1 diabetes and healthy controls with multicolor flow cytometry and t‐distributed Stochastic Neighbor‐Embedded (t‐SNE)‐analyses. IL‐7R expression of monocytes rapidly increased in vitro and was boosted through LPS. In the presence of IL‐7, we detected lower monocyte IL‐7R expression in type 1 diabetes patients as compared to healthy controls. This difference was most evident for the subset of nonclassical monocytes, which increased after IL‐7 stimulation. t‐SNE analyses revealed IL‐7‐dependent differences in monocyte subset distribution and expression of activation and maturation markers (i.e., HLA‐DR, CD80, CD86, CD40). Notably, monocyte CD40 expression increased considerably by IL‐7 and CD40/IL‐7R co‐expression differed between patients and controls. This study shows the unique effects of IL‐7 on monocyte phenotype and functions. Lower IL‐7R expression on IL‐7‐induced CD40_high monocytes and impaired IL‐7 response characterize monocytes from patients with type 1 diabetes.

The common gamma chain (γ_c) contributes to the formation of different cytokine receptors [e.g., IL-2 receptor (IL-2R), IL-7R, and IL-15R], which are important for generation of self-reactive T-cells in autoimmune diseases, like in type 1 diabetes (T1D). Whereas, the roles of membrane and soluble IL-2Rα and IL-7Rα variants in T1D disease pathogenesis are well-described, effects of γ_c expression and availability for dependent receptors remain elusive. We investigated expression of the γ_c and dependent receptors on T-cells and soluble γ_c concentrations in serum from patients with T1D (n = 34) and healthy controls (n = 27). Effector T-cell cytokines as well as IL-2, IL-7, and IL-15 induced STAT5 phosphorylation were analyzed to determine functional implications of differential γ_c expression of CD4⁺ T-cell subsets classified by t-distributed Stochastic Neighbor Embedding (t-SNE) analyses. We found increased γ_c and IL-7Rα expression of CD4⁺ T-cells from T1D patients as compared to controls. t-SNE analyses assigned differential expression to subsets of memory T-cells co-expressing γ_c and IL-7Rα. Whereas, γ_c expression was positively correlated with IL-2Rα in memory T-cells from healthy controls, no dependency was found for patients with T1D. Similarly, the effector T-cell cytokine, IL-21, correlated inversely with γ_c expression in healthy controls, but not in T1D patients. Finally, T1D patients with high γ_c expression had increased proportions of IL-2 sensitive pSTAT5⁺ effector T-cells. These results indicated aberrantly high γ_c expression of T-cells from T1D patients with implications on dependent cytokine receptor signaling and effector T-cell cytokine production.