Characterization of proinsulin-specific regulatory T cells in type 1 diabetes at different ages of onset

Background and Objectives Regulatory T cells (Tregs) play an important role in maintaining tolerance to self-antigens. Defects in the frequency and function of polyclonal Tregs have been reported in type 1 diabetes (T1D). However, characteristics of proinsulin (PI)-specific Tregs in human T1D have not yet been explored. Therefore, we aimed to characterize PI-specific Tregs in two distinct pathophysiological subtypes of T1D, juvenile-onset T1D (JOT1D) and adult-onset T1D (AOT1D), distinguished by the age of onset. Methods Peripheral blood mononuclear cells of the recruited subjects were stimulated in vitro with PI-derived peptides. PI-specific Tregs were characterized by flow cytometry using the combination of markers CD25, CD137, FOXP3 and CD45RA. Results Firstly, we observed similar frequencies of polyclonal Tregs in the T1D (n = 25) and healthy control (HC) (n = 20) subjects (P = 0.96), with a positive correlation between age and frequency of polyclonal Tregs (r = +0.35, P = 0.04). While the frequency of polyclonal Tregs was higher in AOT1D group (P = 0.02), both JOT1D (n = 14) and AOT1D groups (n = 11) had a comparable frequency of PI-specific Tregs in their peripheral blood. The frequency of PI-specific memory Tregs was significantly high in both the JOT1D (P = 0.02) and AOT1D (P = 0.009) groups compared to their respective HC groups (n = 10). Finally, we observed no significant difference in the expression of FOXP3 and IL-2 receptor in PI-specific Tregs in all the groups. Conclusions Unlike polyclonal Tregs, both T1D subtypes harbor comparable frequencies of PI-specific Tregs. Chronic antigen presentation results in a distinct memory-like phenotype of PI-specific Tregs in these subjects irrespective of the age of disease onset.