Type I Interferon Therapy Limits CNS Autoimmunity by Inhibiting CXCR3-Mediated Trafficking of Pathogenic Effector T Cells

Abstract

SUMMARY Type I interferons (IFNs) have therapeutic potential in CNS autoimmune diseases, such as uveitis, but the molecular mechanisms remain unclear. Using a T cell-transfer model of experimental autoimmune uveitis (EAU), we found that IFN-α/β treatment inhibited the migration of IFN-γ-producing pathogenic CD4+ T cells to effector sites. IFN-α/β upregulated the expression of the cognate ligands CXCL9, CXCL10, and CXCL11, causing ligand-mediated downregulation of CXCR3 expression and effector T cell retention in the spleen. Accordingly, type I IFN did not alter EAU progression in CXCR3−/− mice. In uveitis patients, disease exacerbations correlated with reduced serum IFN-α concentrations. IFN-α/β reduced CXCR3 expression and migration by human effector T cells, and these parameters were associated with the therapeutic efficacy of IFN-α in uveitis patients. Our findings provide insight into the molecular basis of type I IFN therapy for CNS autoimmune diseases and identify CXCR3 as a biomarker for effective type I IFN immunotherapy.

Induction of antigen-specific Treg cells in treating autoimmune uveitis via bystander suppressive pathways without compromising anti-tumor immunity

“…Eyes were dissected, digested in RPMI containing 10% FBS and 1 mg/ml collagenase D (Sigma-Aldrich) and then incubated for 1 hr at 37°C. Single cell suspension was prepared for flow cytometry analysis as described [19]. Eyes from naïve mice did not yield leukocytes for analysis.…”

“…Cells were stained using anti-mouse CD3 (17A2), CD4 (GK1.5), CD8a (53-6.7), CD25 (PC61), Foxp3 (FJK-16s), CD11b (M1/70), CD11c (N418), Gr1 (RB6-8C5) Ab or isotype-control Ab. For intracellular cytokine staining with anti-IFN-g (XMG1.2) and anti-IL-17A (TC11-18H10) Abs, cells were incubated for 4 hr with PMA (50 ng/ml, Sigma) and ionomycin (500 ng/ml, Sigma) in the presence of Golgi-Stop (BD) as described [19]. Data were acquired and analyzed using a BD LSRFortessa analyzer and FlowJo software (Tree Star Inc).…”

“…Cells were isolated from the spleens and draining lymphoid nodes (dLNs) 28 days after EAU induction and stimulated with IRBP or soluble aCD3 (17A2)/aCD28 (37.51) Abs in serum-free medium for 48 hr [19,22]. Cytokines were measured from the cultured supernatants by ELISA: IFN-g, IL-17A, TNF-a, and IL-6 (eBioscience).…”

“…Induction of melanoma B16 melanoma cells (ATCC, RRID: CVCL_0604) were cultured in DMEM supplemented with 10% FBS. Melanoma [19,33] was induced in naïve mice or EAU mice (day 40 post-immunization) by subcutaneously injection of tumor cells (0.5 £ 10 6 cells/mouse), and mice were monitored daily for survival, tumor growth and tumor size. At the end of the experiment, tissues were harvested and cells isolated from the spleens and tumors were assessed for FCM.…”

“…Experimental autoimmune uveitis (EAU) is a classical model of human uveitis and can be induced in mice by immunization with retinal antigen, interphotoreceptor retinoid-binding protein (IRBP) [17][18][19].…”