Abstract
The establishment of an “interferon (IFN) signature” to subset SLE patients on disease severity has led to therapeutics targeting IFNα. Here, we investigate IFN signaling in SLE using multiplexed protein arrays and single cell cytometry by time of flight (CyTOF). First, the IFN signature for SLE patients (n=81) from the Stanford Lupus Registry is determined using fluidigm qPCR measuring 44 previously determined IFN-inducible transcripts. IFN-high (IFN-H) patients have increased SLE criteria and renal/CNS/immunologic involvement, and increased autoantibody reactivity against spliceosome-associated antigens. CyTOF analysis is performed on non-stimulated and stimulated (IFNα, IFNγ, IL-21) PBMCs from SLE patients (n=25) and HCs (n=9) in a panel identifying changes in phosphorylation of intracellular signaling proteins (pTOF). Another panel is utilized to detect changes in intracellular cytokine (ICTOF) production in non-stimulated and stimulated (PMA/ionomycin) PBMCs from SLE patients (n=31) and HCs (n=17). Bioinformatic analysis by MetaCyto and OMIQ reveal phenotypic changes in immune cell subsets between IFN-H and IFN-low (IFN-L) patients. Most notably, IFN-H patients exhibit increased STAT1/3/5 phosphorylation downstream of cytokine stimulation and increased phosphorylation of non-canonical STAT proteins. These results suggest that IFN signaling in SLE modulates STAT phosphorylation, potentially uncovering possible targets for future therapeutic approaches.
Highlights
Systemic Lupus Erythematosus (SLE) is a chronic autoimmune disease that carries significant clinical burden for patients including organ damage and death [1–3]
We find that IFN-H patients have more severe disease and increased levels of autoantibodies against spliceosome associated antigens compared to IFN-L patients
We apply highly multiplex single cell mass cytometry to elucidate the effects of the IFN signature on critical SLE-associated cytokines across multiple immune cell subsets
Summary
Systemic Lupus Erythematosus (SLE) is a chronic autoimmune disease that carries significant clinical burden for patients including organ damage and death [1–3]. Transcript profiling of peripheral blood mononuclear cells (PBMCs) [5] and immune cell subsets [6] from SLE patients demonstrate the presence of an IFNainducible gene expression signature in a subset of patients that correlates with clinical SLE criteria and autoantibody production [7]. Recent single cell RNA-seq analysis of kidney biopsies from lupus nephritis (LN) patients reveals an IFN response signature in infiltrating immune cells that correlates with the same signature in the peripheral blood [8, 9]. These studies establish the IFN signature as pathogenic in SLE and support its use to subset patients as IFN-high (IFN-H) or IFN-low (IFN-L). How IFN signaling interacts with other immune pathways in SLE remains unclear
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