Metabolic preconditioning in CD4+ T cells restores inducible immune tolerance in lupus-prone mice
Christopher S. Wilson, Blair T. Stocks, Emilee M. Hoopes, Jillian P. Rhoads, Kelsey L. McNew, Amy S. Major, Daniel J. Moore
Christopher S. Wilson, Blair T. Stocks, Emilee M. Hoopes, Jillian P. Rhoads, Kelsey L. McNew, Amy S. Major, Daniel J. Moore
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Research Article Transplantation

Metabolic preconditioning in CD4+ T cells restores inducible immune tolerance in lupus-prone mice

Abstract

Autoimmune disease has presented an insurmountable barrier to restoration of durable immune tolerance. Previous studies indicate that chronic therapy with metabolic inhibitors can reduce autoimmune inflammation, but it remains unknown whether acute metabolic modulation enables permanent immune tolerance to be established. In an animal model of lupus, we determined that targeting glucose metabolism with 2-deoxyglucose (2DG) and mitochondrial metabolism with metformin enables endogenous immune tolerance mechanisms to respond to tolerance induction. A 2-week course of 2DG and metformin, when combined with tolerance-inducing therapy anti-CD45RB, prevented renal deposition of autoantibodies for 6 months after initial treatment and restored tolerance induction to allografts in lupus-prone mice. The restoration of durable immune tolerance was linked to changes in T cell surface glycosylation patterns, illustrating a role for glycoregulation in immune tolerance. These findings indicate that metabolic therapy may be applied as a powerful preconditioning to reinvigorate tolerance mechanisms in autoimmune and transplant settings that resist current immune therapies.

Authors

Christopher S. Wilson, Blair T. Stocks, Emilee M. Hoopes, Jillian P. Rhoads, Kelsey L. McNew, Amy S. Major, Daniel J. Moore

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Figure 4

A cyclosporine sensitive element modulates immune changes seen with aCD45RB and is defective in SLE123 CD4+ T cells.

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A cyclosporine sensitive element modulates immune changes seen with aCD4...
(A) TRANSFAC analysis of the metabolic genes regulated by aCD45RB in B6 mice revealed ATF-1 and CREBP likely regulated these genes. (B) As ATF-1 and CREB share similar binding motifs, we assessed the nuclear import of both CREB and ATF-1 in aCD45RB-treated and untreated CD4+ T cells from B6 and SLE. Briefly, nuclei from purified CD4+ T cells were extracted and stained for total CREB and ATF-1. The nuclear localized amount of each protein was measured by flow cytometry. No significant difference was noted in any of the conditions regardless of strain or treatment. n = 8 per group, 9- to 12-week-old female mice. (C and D) The transcriptional activity of CREB can be modified by translocation of CRTCs. Nuclear staining demonstrated translocation of CRTC2 to the nucleus following 15 minutes of aCD45RB stimulation only in isolated nuclei from B6 CD4+ T cells. SLE123 mice did not demonstrate the same response. Quantified in D. Representative of 3 repeats with at least 3 biologic replicates per group, utilizing 9- to 12-week-old female mice. (E) The translocation of CRTC proteins is controlled by the phosphatase calcineurin and the SIK, which control nuclear import and export respectively. (F and G) Pretreatment of B6 CD4+ T cells with calcineurin inhibitor, cyclosporine A, but not the NFAT-specific inhibitor, VIVIT (a major target of calcineurin), prevented the metabolic changes induced by aCD45RB. Cyclosporine prevented the downregulation of glucose uptake and changes in mitochondrial dynamics. The NFAT inhibitor VIVIT had no effect on the metabolic changes in glucose uptake or mitochondrial Δψ induced by aCD45RB. Analyzed by 2-way ANOVA and Tukey’s multiple comparison test. n = 5 per each treatment group, 9- to 12-week-old female mice.