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 3

Anti-CD45RB drives transcriptional changes in metabolism in B6 CD4+ T cells, which are resisted by SLE123 mice.

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Anti-CD45RB drives transcriptional changes in metabolism in B6 CD4+ T ce...
(A) CD4+ T cells were isolated from B6 and SLE123 mice that had been treated with a standard 7-day course of aCD45RB or left untreated (n = 3, 9- to 12-week-old female SLE or B6 mice). RNA-Seq was performed and analyzed for differential gene expression. The volcano plot demonstrates regulation of a subset of genes in response to aCD45RB only in B6 mice, while SLE123 mice largely resisted transcriptional changes. (B) KEGG analysis of the downregulated genes in B6 mice revealed metabolic pathways to have the highest gene ratio of regulated pathways. (C) Genes in this KEGG term could be classified broadly into 2 categories that impact both glucose metabolism and mitochondrial processes. (Full gene list available in Supplemental Figure 1B.) (D and E) Analysis of glucose uptake using a fluorescent glucose analog, 2NBDG, revealed a decrease in glucose uptake in CD4+ T cells only from B6 mice treated with CD45RB, which was resisted by SLE123 mice. This corroborated the observation of downregulation of genes associated with glucose metabolism. The percentage of CD4+ T cells with high glucose uptake is quantified in E. (F and G) Mitochondrial membrane potential, a measure of mitochondrial activity, was assessed in both B6 and SLE123 CD4+ T cells by flow cytometry utilizing MitoredCMXRos. This analysis revealed increased mitochondrial membrane potential (Δψ) in CD4+ T cells from B6 mice treated with aCD45RB. SLE123 mice resisted these changes and demonstrated a decrease in membrane potential. The median fluorescence intensity of MitoredCMXRos from F is quantified in G. The analyses in D–G were repeated at least 7 times with 3 or more 9- to 12-week-old female SLE123 or B6 mouse replicates in each group. Analyzed using a 1-way ANOVA and Tukey’s multiple comparison test.