Cell therapies for autoimmune diseases using tolerogenic dendritic cells (tolDC) have been promisingly explored. A major stumbling block has been generating stable tolDC, with low risk of converting to mature immunogenic DC (mDC), exacerbating disease. mDC induction involves a metabolic shift to lactate production from oxidative phosphorylation (OXPHOS) and β-oxidation, the homeostatic energy source for resting DC. Inhibition of glycolysis through the administration of 2-deoxy glucose (2-DG) has been shown to prevent autoimmune disease experimentally but is not clinically feasible. We show here that treatment of mouse bone marrow-derived tolDC ex vivo with low-dose 2-DG (2.5 mM) (2-DGtolDC) induces a stable tolerogenic phenotype demonstrated by their failure to engage lactate production when challenged with mycobacterial antigen (Mtb). ~ 15% of 2-DGtolDC express low levels of MHC class II and 30% express CD86, while they are negative for CD40. 2-DGtolDC also express increased immune checkpoint molecules PDL-1 and SIRP-1α. Antigen-specific T cell proliferation is reduced in response to 2-DGtolDC in vitro. Mtb-stimulated 2-DGtolDC do not engage aerobic glycolysis but respond to challenge via increased OXPHOS. They also have decreased levels of p65 phosphorylation, with increased phosphorylation of the non-canonical p100 pathway. A stable tolDC phenotype is associated with sustained SIRP-1α phosphorylation and p85-AKT and PI3K signalling inhibition. Further, 2-DGtolDC preferentially secrete IL-10 rather than IL-12 upon Mtb-stimulation. Importantly, a single subcutaneous administration of 2-DGtolDC prevented experimental autoimmune uveoretinitis (EAU) in vivo. Inhibiting glycolysis of autologous tolDC prior to transfer may be a useful approach to providing stable tolDC therapy for autoimmune/immune-mediated diseases.

使用耐受性树突状细胞（tolDC）的自身免疫性疾病的细胞疗法已经得到了有前途的探索。一个主要的绊脚石一直在产生稳定的tolDC，转化为成熟的免疫原性DC（mDC）的风险较低，从而加剧了疾病。mDC诱导涉及从氧化磷酸化（OXPHOS）和β-氧化（用于静息DC的稳态能量源）向乳酸生成的代谢转变。已经显示通过施用2-脱氧葡萄糖（2-DG）来抑制糖酵解可以预防自身免疫性疾病，但是在临床上并不可行。我们在这里显示了低剂量的2-DG（2.5 mM）（2-DGtolDC）的小鼠骨髓来源的tolDC的离体治疗诱导了稳定的耐受性表型，表现为当它们受到分枝杆菌抗原（Mtb）攻击时无法参与乳酸产生）。约15％的2-DGtolDC表达低水平的II类MHC，而30％则表达CD86，而CD40阴性。2-DGtolDC还表达增加的免疫检查点分子PDL-1和SIRP-1α。在体外对2-DGtolDC的反应降低了抗原特异性T细胞的增殖。Mtb刺激的2-DGtolDC不参与有氧糖酵解，但通过增加OXPHOS来应对挑战。它们还具有降低的p65磷酸化水平，以及非规范性p100途径的磷酸化增加。稳定的tolDC表型与持续的SIRP-1α磷酸化以及p85-AKT和PI3K信号传导抑制有关。此外，2-DGtolDC在Mtb刺激时优先分泌IL-10而不是IL-12。重要的是，一次皮下注射2-DGtolDC可以在体内预防实验性自身免疫性葡萄膜视网膜炎（EAU）。