5a, displaying an overlay of the FOXP3+CD25+ expression after expansion of sorted CD4+CD25+CD127lo/− cells and CD4+CD25− cells, from the same individual. Fig. 5b shows the FOXP3+CD25+

expression after expansion of sorted CD4+CD25− cells and Fig. 5c of sorted CD4+CD25+CD127lo/− cells. Cumulative data of the expansion of sorted CD4+CD25+CD127lo/− and CD4+CD25− confirmed that a higher percentage of sorted and expanded CD4+CD25+CD127lo/− was FOXP3 positive (p<0.001, Fig. 5d). Moreover, expanded CD4+CD25+CD127lo/− had a higher mean fluorescent intensity (MFI) of FOXP3, both expressed as geometrical mean (p<0.001, Fig. 5e) and mean (p<0.001, Fig. 5f), compared to expanded CD4+CD25−. The percentage see more of FOXP3 expressing cells in the expanded CD4+CD25+CD127lo/− cultures did not differ significantly between this website T1D, healthy and high-risk individuals, even if T1D visually appeared to be higher (Fig. 6a). Post-expansion, CD4+CD25+CD127lo/− from T1D in contrast tended to display higher FOXP3 MFI, both expressed as geometrical mean (p=0.05, Fig. 6b) and mean (p=0.05, Fig. 6c), compared to healthy individuals. Expansion of CD4+CD25− T-cells did not yield any significant differences in the percentage of FOXP3 expressing cells or in FOXP3 MFI, neither expressed as geometrical mean nor

mean (data not shown). No differences, in regard to age distribution, were seen in composition of the studied T-cell phenotypes in healthy

or high-risk individuals (data not shown). Neither was fold increase of CD4+CD25+CD127lo/− T-cells correlated to age among the high-risk, healthy or T1D individuals, nor in the total study population as a group (data not shown). The high-risk group was further split in regard to development of T1D after inclusion in the study as well as split for treatment with nicotinamide or placebo but showed no differences in T-cell composition (data PD184352 (CI-1040) not shown). During recent years, the interest for Tregs has increased, and their role and function have been thoroughly scrutinized in a plethora of studies. While their suppressive functions and importance in maintaining immune homoeostasis in experimental models are generally acknowledged, their actual involvement in human autoimmune disease is more disputed and reported findings are non-unanimous. The development of T1D is associated with an imbalance in the immune system connected to an autoimmune attack on the insulin producing β-cells. A vast number of studies have identified pieces of the immunological puzzle of T1D, seeking to unravel the secret of the autoimmune riddle. However the origin, the failing regulatory mechanism rendering subjects non-tolerant to self remains elusive. Therefore, all pieces that can be added to this puzzle will be important for the picture to appear.