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 5

CD4 surface expression of CD45RB and global glycosylation are altered in SLE123 mice.

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CD4 surface expression of CD45RB and global glycosylation are altered in...
(A and B) Cell surface expression of CD45RB and total CD45 were measured by flow cytometry on CD4+ T cells. SLE123 demonstrated a downregulation of CD45RB on the cell surface as compared with B6 CD4+ T cells. There was no difference in pan-CD45 expression. Quantified in B. (C) CD45 is composed of an intracellular region that controls cytoskeletal binding and its phosphatase activity. The extracellular domain is composed of a region with fibronectin repeats that is heavily N-glycosylated. An alternatively spliced region is heavily O-glycosylated, and this region imparts unique functions to each CD45 isoform. Additionally, a sialic acid residue on the B portion of this region is essential for the binding of therapeutic aCD45RB. (D and E) We utilized lectins to detect the level of α-2,3–linked sialylation (MALII), O-linked glycosylation (Jacalin), and N-linked glycosylation (PHA-L) in CD4+ T cells from B6 and SLE123 mice. We determined SLE123 CD4+ T cells had reduced levels of α-2,3–linked sialic acids and O-glycosylation and an increase in N-glycosylation. Quantified in E. (F) Utilizing an antibody that detects a desialylated form of anti-CD45RB, we determined SLE123 CD4+ T cells possessed increased binding of this antibody compared with B6. (G and H) To determine the binding of therapeutic aCD45RB to B6 and SLE123 CD4+ T cells, we incubated splenocytes from both strains with aCD45RB, followed by an anti–rat IgG2a antibody conjugated to phycoerythrin. Flow cytometry demonstrated CD4+ T cells from SLE123 mice had reduced binding of the therapeutic antibody. Quantified in H. Analyzed using a Student’s t test. Representative data of at least 4 experimental repeats, with at least 3 biologic replicates of 9- to 12-week-old mice in each group.