Stage Dependent Aberrant Regulation of Cytokine-STAT Signaling in Murine Systemic Lupus Erythematosus

The Baxter Laboratory of Genetic Pharmacology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA.
PLoS ONE (Impact Factor: 3.23). 02/2009; 4(8):e6756. DOI: 10.1371/journal.pone.0006756
Source: PubMed

ABSTRACT Systemic lupus erythematosus (SLE) is a complex autoimmune disease of unknown etiology that involves multiple interacting cell types driven by numerous cytokines and autoimmune epitopes. Although the initiating events leading to SLE pathology are not understood, there is a growing realization that dysregulated cytokine action on immune cells plays an important role in promoting the inflammatory autoimmune state. We applied phospho-specific flow cytometry to characterize the extent to which regulation of cytokine signal transduction through the STAT family of transcription factors is disturbed during the progression of SLE. Using a panel of 10 cytokines thought to have causal roles in the disease, we measured signaling responses at the single-cell level in five immune cell types from the MRLlpr murine model. This generated a highly multiplexed view of how cytokine stimuli are processed by intracellular signaling networks in adaptive and innate immune cells during different stages of SLE pathogenesis. We report that robust changes in cytokine signal transduction occur during the progression of SLE in multiple immune cell subtypes including increased T cell responsiveness to IL-10 and ablation of Stat1 responses to IFNalpha, IFNgamma, IL-6, and IL-21, Stat3 responses to IL-6, Stat5 responses to IL-15, and Stat6 responses to IL-4. We found increased intracellular expression of Suppressor of Cytokine Signaling 1 protein correlated with negative regulation of Stat1 responses to inflammatory cytokines. The results provide evidence of negative feedback regulation opposing inflammatory cytokines that have self-sustaining activities and suggest a cytokine-driven oscillator circuit may drive the periodic disease activity observed in many SLE patients.

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Available from: Matthew B Hale, Sep 27, 2015
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    • "In our study, plasma levels of proinflammatory cytokines IL-1β, IL-12(p70), IFN-γ, and IL-6 are significantly augmented in SLE patients with inactive or middle disease compared with controls. In SLE it has been suggested that aberrant control of immune cell responses to inflammatory cytokines may disrupt the delicate balance between immunity and self-reactivity [2, 3, 88]. IL-6 has been studied in human SLE, and IL-6 was found to reflect disease activity [19, 89–93]. "
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    ABSTRACT: Kinases have been implicated in the immunopathological mechanisms of Systemic Lupus Erythematosus (SLE). v-akt murine-thymoma viral-oncogene-homolog 1 (AKT1) and mitogen-activated-protein-kinase 1 (MAPK1) gene expressions in peripheral mononuclear cells from thirteen SLE patients with inactive or mild disease were evaluated using quantitative real-time reverse-transcription polymerase-chain-reaction and analyzed whether there was any correlation with T-helper (Th) transcription factors (TF) gene expression, cytokines, and S100A8/S100A9-(Calprotectin). Age- and gender-matched thirteen healthy controls were examined. AKT1 and MAPK1 expressions were upregulated in SLE patients and correlated with Th17-(Retinoic acid-related orphan receptor (ROR)-C), T-regulatory-(Treg)-(Transforming Growth Factor Beta (TGFB)-2), and Th2-(interleukin (IL)-5)-related genes. MAPK1 expression correlated with Th1-(IL-12A, T-box TF-(T-bet)), Th2-(GATA binding protein-(GATA)-3), and IL-10 expressions. IL-10 expression was increased and correlated with plasma Tumor Necrosis Factor (TNF)-α and Th0-(IL-2), Th1-(IL-12A, T-bet), GATA3, Treg-(Forkhead/winged-helix transcription factor- (FOXP)-3), and IL-6 expressions. FOXP3 expression, FOXP3/RORC, and FOXP3/GATA3 expression ratios were increased. Plasma IL-1β, IL-12(p70), Interferon-(IFN)-γ, and IL-6 cytokines were augmented. Plasma IL-1β, IL-6, IL-2, IFN-γ, TNF-α, IL-10, and IL-13 correlated with C-reactive protein, respectively. Increased Calprotectin correlated with neutrophils. Conclusion, SLE patients presented a systemic immunoinflammatory activity, augmented AKT1 and MAPK1 expressions, proinflammatory cytokines, and Calprotectin, together with increased expression of Treg-related genes, suggesting a regulatory feedback opposing the inflammatory activity.
    Mediators of Inflammation 10/2012; 2012(11):495934. DOI:10.1155/2012/495934 · 3.24 Impact Factor
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    • "Flow cytometry analysis of the presence of myeloid cells showed that pSAP-treated lupus mice exhibited a notable decrease in the number of F4/80+ macrophages (Fig. 6A), but there was no significant decrease in the number of CD11c+ dendritic cells (data not shown), suggesting that macrophages were the key cells that was influenced by pSAP treatment. As a set of inflammatory markers mainly secreted by macrophages were expressed in kidneys following glomerular immune complex deposition [8], [18], [19], further studies using ELISA analysis allowed us to determine several key markers in kidneys of mice. It was found that TNF-α, IL-1β, IL-6, IL-12 and MCP-1, which were upregulated in kidneys of ALD-DNA-induced lupus mice, were decreased in the pSAP-treated lupus mice (Fig. 6B). "
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    ABSTRACT: Our previous study revealed that administration of syngeneic female BALB/c mice with excessive self activated lymphocyte-derived DNA (ALD-DNA) could induce systemic lupus erythematosus (SLE) disease, indicating that overload of self-DNA might exceed normal clearance ability and comprise the major source of autoantigens in lupus mice. Serum amyloid P component (SAP), an acute-phase serum protein with binding reactivity to DNA in mice, was proved to promote the clearance of free DNA and prevent mice against self-antigen induced autoimmune response. It is reasonable to hypothesize that SAP treatment might contribute to alleviation of SLE disease, whereas its role in ALD-DNA-induced lupus nephritis is not fully understood. The ratios of SAP to DNA significantly decreased and were negatively correlated with the titers of anti-dsDNA antibodies in ALD-DNA-induced lupus mice, indicating SAP was relatively insufficient in lupus mice. Herein a pcDNA3-SAP plasmid (pSAP) was genetically constructed and intramuscularly injected into BALB/c mice. It was found that SAP protein purified from the serum of pSAP-treated mice bound efficiently to ALD-DNA and inhibited ALD-DNA-mediated innate immune response in vitro. Treatment of ALD-DNA-induced lupus mice with pSAP in the early stage of SLE disease with the onset of proteinuria reversed lupus nephritis via decreasing anti-dsDNA autoantibody production and immune complex (IC) deposition. Further administration of pSAP in the late stage of SLE disease that had established lupus nephritis alleviated proteinuria and ameliorated lupus nephritis. This therapeutic effect of SAP was not only attributable to the decreased levels of anti-dsDNA autoantibodies, but also associated with the decreased infiltration of lymphocytes and the reduced production of inflammatory markers. These results suggest that SAP administration could effectively alleviated lupus nephritis via modulating anti-dsDNA antibody production and the inflammation followed IC deposition, and SAP-based intervening strategy may provide new approaches for treating SLE disease.
    PLoS ONE 07/2011; 6(7):e22659. DOI:10.1371/journal.pone.0022659 · 3.23 Impact Factor
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    • "Dysregulated cytokine signaling in lymphocytes is thought to contribute to autoimmune disease progression [19 –21]. An example of this, dysregulation was shown in a detailed longitudinal study of cytokine signaling in lymphocyte subsets using an MRL/lpr mouse model of SLE [22]. Here, the authors stimulated splenocytes ex vivo with a panel of 10 cytokines to evoke Janus kinases (Jak)/signal transducers and activators of transcription (Stat) signaling at different stages of SLE pathogenesis, and in normal mice. "
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    ABSTRACT: Autoimmune diseases are complex and heterogeneous in nature and show varying responses to therapeutic treatment. A more accurate biological characterization of individual patients would assist in disease classification, prognosis, and treatment decisions. This characterization ideally would incorporate cellular, biochemical, and molecular information that contribute to the inflammatory state. The advent of single-cell network profiling (SCNP) using phospho-flow multiparametric flow cytometry allows insight into the complexity of signaling networks in various cell types. Here, we describe the potential of SCNP to inform on the biological characterization of autoimmune disease, the application in clinical medicine, and the utility in drug development.
    Autoimmunity 11/2010; 43(7):550-9. DOI:10.3109/08916931003674774 · 2.71 Impact Factor
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