Moreover, emerging evidence supports a direct correlation between DC numbers and the proliferation rate of peripheral Treg. Thus, Fms-like tyrosine kinase 3 ligand (Flt3L) treatment, which results in the in vivo expansion of classical DC (cDC) 11 leads to a concomitant increase in peripheral Treg 12, 13. Furthermore, it was recently demonstrated that the conditional ablation of cDC from otherwise intact animals results in reduced numbers and impaired homeostatic proliferation of peripheral Treg 13. Here, we readdressed the
role of cDC in the maintenance of peripheral Treg focusing on the role of CD80/86 costimulation. Using constitutive and conditional cDC ablation strategies, we established that peripheral Treg maintenance critically
depends on the presence of cDC expressing CD80/86. Surprisingly however and defying earlier notions 13, 14, the reduction of Treg in animals BMS-907351 clinical trial lacking cDC as such was not inherently associated with lymphocyte activation. Rather than resulting from a tolerance Afatinib concentration failure, the autoinflammatory signatures reported for cDC-deficient mice are thus a consequence of the nonmalignant myeloproliferative disorder these animals develop. We and others recently reported that animals that constitutively lack cDC (CD11c-DTA mice) display normal percentages and numbers of thymic Foxp3+ Treg 14, 15, thereby establishing that DC are dispensable for the generation of nTreg. Moreover, CD11c-DTA mice retained functional peripheral Treg 15. However, closer examination of the blood circulation and LN of cDC-deficient animals and comparison to their littermate controls revealed
a twofold reduction in the frequencies of Treg out of total CD4+ T cells, whose numbers are unaltered 15 (Fig. 1A). This reduction of peripheral Foxp3+ Treg was also observed upon conditional cDC ablation, as achieved through repetitive diphtheria toxin (DTx) treatment of [CD11c-DTR>WT] BM chimeras (Fig. 1B) 16, thereby confirming recent reports that established the critical role of cDC Adenosine in promoting the homeostatic Treg proliferation 13, 17. Re-examination of Treg frequencies in cDC-deficient animals by staining for both Foxp3 and CD25 revealed a twofold reduction of Foxp3+CD25+ (double positive) Treg in all organs tested, including the spleen (Fig. 1C–E). Interestingly though, the decrease of splenic Foxp3+CD25+ Treg was uniquely associated with a concomitant elevation in the frequencies of Foxp3+CD25− (single positive) cells out of CD4+ T cells (Fig. 1E). This finding explains the reason why the splenic Foxp3+ T-cell compartment of cDC-deficient CD11c:DTA mice had, in the previous studies, appeared unaffected 14, 15. Collectively, these data establish that although cDC are not required for the generation of nTreg in the thymus, they are – in agreement with recent reports 13, 17 – critically involved in the maintenance of peripheral Foxp3+CD25+ Treg.