Regulatory T-Cell Augmentation or Interleukin-17 Inhibition Prevents Calcineurin Inhibitor–Induced Hypertension in Mice

Abstract

The immunosuppressive calcineurin inhibitors cyclosporine A and tacrolimus alter T cell subsets and can cause hypertension, vascular dysfunction, and renal toxicity. We and others have reported that cyclosporine A and tacrolimus decrease anti-inflammatory regulatory T cells and increase pro-inflammatory interleukin-17-producing T cells, therefore we hypothesized that inhibition of these effects using non-cellular therapies would prevent the hypertension, endothelial dysfunction, and renal glomerular injury induced by calcineurin inhibitor therapy. Daily treatment of mice with cyclosporine A or tacrolimus for 1 week significantly decreased CD4+/FoxP3+ regulatory T cells in the spleen and lymph nodes, as well as induced hypertension, vascular injury and dysfunction, and glomerular mesangial expansion in mice. Daily co-treatment with all-trans retinoic acid, reported to increase regulatory T cells and decrease interleukin-17-producing T cells, prevented all of the detrimental effects of cyclosporine A and tacrolimus. All-trans retinoic acid also increased regulatory T cells and prevented the hypertension, endothelial dysfunction, and glomerular injury in genetically modified mice that phenocopy calcineurin inhibitor-treated mice (FKBP12-Tie2 KO). Treatment with an interleukin-17 neutralizing antibody also increased regulatory T cell levels and prevented the hypertension, endothelial dysfunction, and glomerular injury in cyclosporine A-treated and tacrolimus-treated mice as well as FKBP12-Tie2 KO mice, while an isotype control had no effect. Augmenting regulatory T cells and/or inhibiting interleukin-17 signaling using non-cellular therapies prevents the cardiovascular and renal toxicity of calcineurin inhibitors in mice.

T Cells and Acute Kidney Injury: A Two-Way Relationship

“…Clinically relevant approaches preventing T H17 responses (i.e., EPO receptor agonism and Orai1 inhibition) effectively halted kidney disease in murine models. Moreover, targeting T H17 effector molecule, IL-17, prevented calcineurin inhibitor related renal fibrosis (70). Taken together, these findings justify further studies targeting T H17 responses to prevent CKD.…”

Impaired mitochondrial calcium uptake caused by tacrolimus underlies beta-cell failure

“…Further studies are necessary to decipher in detail the exact processes linking tacrolimus pharmacodynamics to the observed effects on ER and mitochondrial Ca 2+ reported here for the first time. Potential candidates, currently being investigated in our laboratory, include the immunophilin FK506 binding protein (FKBP), which forms a complex with tacrolimus [ 57 – 59 ], and ER stress, which has been recently reported in tacrolimus-treated lymphocytes [ 60 ].…”

Myeloid-Derived Suppressor Cells Ameliorate Cyclosporine A–Induced Hypertension in Mice

“…Following blood pressure measures, mice were injected ip with CSA (50 mg/kg/day; Alomone, Israel) or diluent (saline and DMSO, 0.2% final concentration) as described previously. 9, 30–32 Some mice were also given daily i.p. injections of IL-33 (0.5 ug/mouse/day; eBioscience).…”

“…For protein analyses from whole aortic and kidney tissue, tissue lysates were processed and total protein concentration was determined as described previously. 9 The blots were probed for Fibronectin (Abcam), E-selectin (Abcam), collagen (Millipore), calcineurin (Cell Signaling), and β-actin (Sigma) and were identified simultaneously (800 nm and 700 nm wavelengths, respectively) using near-infrared visualization (Odyssey System, LI-COR Biosciences). Densitometry was performed using the Odyssey software.…”

“…9 Three blinded investigators quantitated glomerular injury according to a scale described previously. 9 In brief, 15 glomeruli per tissue section were scored on a 0–4 scale, where 0=normal size, normal mesangium, and no congestion; 1=minimal hypertrophy, mesangial expansion, and/or congestion; 2=mild hypertrophy, mesangial expansion, and/or congestion; 3=moderate hypertrophy, mesangial expansion, and/or congestion; and 4=severe hypertrophy, mesangial expansion, and congestion. Mean glomerular injury index for each group was calculated by [(0 × N0)+(1 × N1)+(2 × N2)+(3 × N3)+(4 × N4)]/15, where N0-N4=number of glomeruli with scores of 0 to 4, respectively, as described previously.…”