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No Alterations in the Frequency of FOXP3⁺ Regulatory T-Cells in Type 1 Diabetes

¹ Department of Pathology, University of Florida, Gainesville, Florida
² Department of Pediatrics, University of Florida, Gainesville, Florida
³ Barbara Davis Center for Childhood Diabetes, University of Colorado Health Sciences Center, Aurora, Colorado

Address correspondence and reprint requests to Mark A. Atkinson, PhD, Department of Pathology, College of Medicine, University of Florida, ARB-R3-128, 1600 SW Archer Rd., Gainesville, FL 32610-0275. E-mail: atkinson{at}ufl.edu

Abbreviations: APC, allophycocyanin; CBC, complete blood count; CD25, -chain of the interleukin-2 receptor complex; FITC, fluorescein isothiocyanate; FOXP3, transcription factor forkhead box P3; GADA, GAD autoantibody; IL, interleukin; MHC, major histocompatibility complex; PE, phycoerythrin; Teff, CD4⁺CD25^– effector T-cell; Treg, CD4⁺CD25⁺FOXP3⁺ regulatory T-cell

Regulatory T-cells (Tregs) play a critical role in maintaining dominant peripheral tolerance. Previous characterizations of Tregs in type 1 diabetes have used antibodies against CD4 and -chain of the interleukin-2 receptor complex (CD25). This report extends those investigations by the addition of a more lineage-specific marker for Tregs, transcription factor forkhead box P3 (FOXP3), in subjects with type 1 diabetes, their first-degree relatives, and healthy control subjects. With inclusion of this marker, two predominant populations of CD4⁺CD25⁺ T-cells were identified: CD4⁺CD25⁺FOXP3⁺ as well as CD4⁺FOXP3^– T-cells expressing low levels of CD25 (CD4⁺CD25^LOWFOXP3^–). In all study groups, the frequency of CD4⁺CD25⁺FOXP3⁺ cells was age independent, whereas CD4⁺CD25^LOWFOXP3^– cell frequencies strongly associated with age. In terms of additional markers for delineating cells of Treg lineage, FOXP3⁺ cells were CD127^– to CD127^LOW whereas CD25⁺ cells were less restricted in their expression of this marker, with CD127 expressed across a continuum of levels. Importantly, no differences were observed in the frequency of CD4⁺CD25⁺FOXP3⁺ T-cells in individuals with or at varying degrees of risk for type 1 diabetes. These investigations suggest that altered peripheral blood frequencies of Tregs, as defined by the expression of FOXP3, are not specifically associated with type 1 diabetes and continue to highlight age as an important variable in analysis of immune regulation.

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