RESUMEN

Gain-of-function mutations that activate the innate immune system can cause systemic autoinflammatory diseases associated with increased IL-1ß production. This cytokine is activated identically to IL-18 by an intracellular protein complex known as the inflammasome; however, IL-18 has not yet been specifically implicated in the pathogenesis of hereditary autoinflammatory disorders. We have now identified an autoinflammatory disease in mice driven by IL-18, but not IL-1ß, resulting from an inactivating mutation of the actin-depolymerizing cofactor Wdr1. This perturbation of actin polymerization leads to systemic autoinflammation that is reduced when IL-18 is deleted but not when IL-1 signaling is removed. Remarkably, inflammasome activation in mature macrophages is unaltered, but IL-18 production from monocytes is greatly exaggerated, and depletion of monocytes in vivo prevents the disease. Small-molecule inhibition of actin polymerization can remove potential danger signals from the system and prevents monocyte IL-18 production. Finally, we show that the inflammasome sensor of actin dynamics in this system requires caspase-1, apoptosis-associated speck-like protein containing a caspase recruitment domain, and the innate immune receptor pyrin. Previously, perturbation of actin polymerization by pathogens was shown to activate the pyrin inflammasome, so our data now extend this guard hypothesis to host-regulated actin-dependent processes and autoinflammatory disease.

Asunto(s)

Actinas/fisiología , Proteínas del Citoesqueleto/metabolismo , Enfermedades Autoinflamatorias Hereditarias/metabolismo , Inflamación/metabolismo , Interleucina-1beta/metabolismo , Proteínas de Microfilamentos/metabolismo , Actinas/química , Animales , Células de la Médula Ósea/citología , Caspasa 1/metabolismo , Caspasas/metabolismo , Ácido Clodrónico/química , Cruzamientos Genéticos , Medios de Cultivo Condicionados/química , Ensayo de Inmunoadsorción Enzimática , Interleucina-18/metabolismo , Lipopolisacáridos/metabolismo , Liposomas/química , Hígado/embriología , Factor Estimulante de Colonias de Macrófagos/metabolismo , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Microscopía Fluorescente , Monocitos/citología , Pirina , Transducción de Señal

RESUMEN

Obesity is associated with infiltration of macrophages into adipose tissue (AT), contributing to insulin resistance and diabetes. However, relatively little is known regarding the origin of AT macrophages (ATMs). We discovered that murine models of obesity have prominent monocytosis and neutrophilia, associated with proliferation and expansion of bone marrow (BM) myeloid progenitors. AT transplantation conferred myeloid progenitor proliferation in lean recipients, while weight loss in both mice and humans (via gastric bypass) was associated with a reversal of monocytosis and neutrophilia. Adipose S100A8/A9 induced ATM TLR4/MyD88 and NLRP3 inflammasome-dependent IL-1ß production. IL-1ß interacted with the IL-1 receptor on BM myeloid progenitors to stimulate the production of monocytes and neutrophils. These studies uncover a positive feedback loop between ATMs and BM myeloid progenitors and suggest that inhibition of TLR4 ligands or the NLRP3-IL-1ß signaling axis could reduce AT inflammation and insulin resistance in obesity.

Asunto(s)

Tejido Adiposo/metabolismo , Macrófagos/metabolismo , Monocitos/metabolismo , Mielopoyesis/fisiología , Obesidad/metabolismo , Animales , Médula Ósea/metabolismo , Proteínas Portadoras/metabolismo , Proliferación Celular/fisiología , Humanos , Inflamasomas/metabolismo , Interleucina-1beta/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Factor 88 de Diferenciación Mieloide/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR , Neutrófilos/metabolismo , Receptores de Interleucina-1/metabolismo , Receptor Toll-Like 4/metabolismo

RESUMEN

Diabetes is a major risk factor for atherosclerosis. Although atherosclerosis is initiated by deposition of cholesterol-rich lipoproteins in the artery wall, the entry of inflammatory leukocytes into lesions fuels disease progression and impairs resolution. We show that diabetic mice have increased numbers of circulating neutrophils and Ly6-C(hi) monocytes, reflecting hyperglycemia-induced proliferation and expansion of bone marrow myeloid progenitors and release of monocytes into the circulation. Increased neutrophil production of S100A8/S100A9, and its subsequent interaction with the receptor for advanced glycation end products on common myeloid progenitor cells, leads to enhanced myelopoiesis. Treatment of hyperglycemia reduces monocytosis, entry of monocytes into atherosclerotic lesions, and promotes regression. In patients with type 1 diabetes, plasma S100A8/S100A9 levels correlate with leukocyte counts and coronary artery disease. Thus, hyperglycemia drives myelopoiesis and promotes atherogenesis in diabetes.

Asunto(s)

Aterosclerosis/patología , Hiperglucemia/patología , Mielopoyesis/fisiología , Animales , Aterosclerosis/metabolismo , Médula Ósea/metabolismo , Médula Ósea/patología , Enfermedad Coronaria/metabolismo , Enfermedad Coronaria/patología , Citocinas/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 1/patología , Glucosa/metabolismo , Productos Finales de Glicación Avanzada/metabolismo , Humanos , Hiperglucemia/metabolismo , Leucocitos/metabolismo , Leucocitos/patología , Leucocitosis/metabolismo , Leucocitosis/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Monocitos/metabolismo , Monocitos/patología , Células Progenitoras Mieloides/metabolismo , Células Progenitoras Mieloides/patología , FN-kappa B/metabolismo , Neutrófilos/metabolismo , Neutrófilos/patología , Receptor para Productos Finales de Glicación Avanzada , Receptores Inmunológicos/metabolismo

RESUMEN

Effective humoral responses to protein antigens require the precise execution of carefully timed differentiation programs in both T and B cell compartments. Disturbances in this process underlie the pathogenesis of many autoimmune disorders, including systemic lupus erythematosus (SLE). Interferon regulatory factor 4 (IRF4) is induced upon the activation of T and B cells and serves critical functions. In CD4(+) T helper cells, IRF4 plays an essential role in the regulation of IL-21 production, whereas in B cells it controls class switch recombination and plasma cell differentiation. IRF4 function in T helper cells can be modulated by its interaction with regulatory protein DEF6, a molecule that shares a high degree of homology with only one other protein, SWAP-70. Here, we demonstrate that on a C57BL/6 background the absence of both DEF6 and SWAP-70 leads to the development of a lupus-like disease in female mice, marked by simultaneous deregulation of CD4(+) T cell IL-21 production and increased IL-21 B cell responsiveness. We furthermore show that DEF6 and SWAP-70 are differentially used at distinct stages of B cell differentiation to selectively control the ability of IRF4 to regulate IL-21 responsiveness in a stage-specific manner. Collectively, these data provide novel insights into the mechanisms that normally couple and coordinately regulate T and B cell responses to ensure tight control of productive T-B cell interactions.

Asunto(s)

Linfocitos B/inmunología , Factores Reguladores del Interferón/inmunología , Linfocitos T/inmunología , Animales , Autoanticuerpos/biosíntesis , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/prevención & control , Diferenciación Celular , Proteínas de Unión al ADN/deficiencia , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/inmunología , Femenino , Factores de Intercambio de Guanina Nucleótido/deficiencia , Factores de Intercambio de Guanina Nucleótido/genética , Factores de Intercambio de Guanina Nucleótido/inmunología , Interleucinas/biosíntesis , Cooperación Linfocítica/inmunología , Trastornos Linfoproliferativos/etiología , Trastornos Linfoproliferativos/genética , Trastornos Linfoproliferativos/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Antígenos de Histocompatibilidad Menor , Proteínas Nucleares/deficiencia , Proteínas Nucleares/genética , Proteínas Nucleares/inmunología , Células Plasmáticas/inmunología

RESUMEN

OBJECTIVE: Neutrophils play a key role in the immune response but can undesirably exacerbate inflammation. High-density lipoproteins (HDL) are antiinflammatory particles, exerting beneficial cardiovascular influences. We determined whether HDL exerts antiinflammatory effects on neutrophils and explored the mechanisms by which these occur. METHODS AND RESULTS: CD11b on activated human neutrophils was significantly attenuated by apolipoprotein A-I (apoA-I) and HDL. The effects of apoA-I were mediated via ABCA1, whereas the effects of HDL were via scavenger receptor BI. Both were associated with a reduction in the abundance of lipid rafts, and a strong correlation between raft abundance and CD11b activation was observed. ApoA-I and HDL reduced neutrophil adhesion to a platelet monolayer under shear flow, as well as neutrophil spreading and migration. ApoA-I also inhibited leukocyte recruitment to the endothelium in an acute in vivo model of inflammation. Finally, infusion of reconstituted HDL in patients with peripheral vascular disease was demonstrated to significantly attenuate neutrophil activation. CONCLUSION: We describe here a novel role for HDL and apoA-I in regulating neutrophil activation using in vitro, in vivo, and clinical approaches. We also show that these effects of HDL and apoA-I involve a mechanism requiring changes in membrane domain content rather than in cholesterol efflux per se.

Asunto(s)

Apolipoproteína A-I/fisiología , Inflamación/inmunología , Lipoproteínas HDL/fisiología , Activación Neutrófila , Animales , Antígeno CD11b/análisis , Adhesión Celular , Movimiento Celular , Colesterol/metabolismo , Humanos , Inflamación/prevención & control , Masculino , Microdominios de Membrana/fisiología , Ratones , Ratones Endogámicos C57BL , Enfermedades Vasculares Periféricas/inmunología , Receptores Depuradores de Clase B , Acetato de Tetradecanoilforbol/farmacología

RESUMEN

Deregulated production of IL-17 and IL-21 plays a key pathogenic role in many autoimmune disorders. A delineation of the mechanisms that underlie the inappropriate synthesis of IL-17 and IL-21 in autoimmune diseases can thus provide important insights into potential therapies for these disorders. Here we have shown that the serine-threonine kinase Rho-associated, coiled-coil-containing protein kinase 2 (ROCK2) becomes activated in mouse T cells under Th17 skewing conditions and phosphorylates interferon regulatory factor 4 (IRF4), a transcription factor that is absolutely required for the production of IL-17 and IL-21. We furthermore demonstrated that ROCK2-mediated phosphorylation of IRF4 regulated the synthesis of IL-17 and IL-21 and the differentiation of Th17 cells. Whereas CD4+ T cells from WT mice activated ROCK2 physiologically under Th17 conditions, CD4+ T cells from 2 different mouse models of spontaneous autoimmunity aberrantly activated ROCK2 under neutral conditions. Moreover, administration of ROCK inhibitors ameliorated the deregulated production of IL-17 and IL-21 and the inflammatory and autoantibody responses observed in these autoimmune mice. Our findings thus uncover a crucial link among ROCK2, IRF4, and the production of IL-17 and IL-21 and support the idea that selective inhibition of ROCK2 could represent an important therapeutic regimen for the treatment of autoimmune disorders.

Asunto(s)

Factores Reguladores del Interferón/inmunología , Factores Reguladores del Interferón/metabolismo , Interleucina-17/metabolismo , Interleucinas/metabolismo , Quinasas Asociadas a rho/metabolismo , Animales , Autoinmunidad/inmunología , Diferenciación Celular/inmunología , Interleucina-17/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Fosforilación , Proteínas Serina-Treonina Quinasas/inmunología , Proteínas Serina-Treonina Quinasas/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismo

RESUMEN

OBJECTIVE: Whereas the anti-inflammatory effects of high-density lipoprotein (HDL) on endothelial cells are well described, such effects on monocytes are less studied. METHODS AND RESULTS: Human monocytes were isolated from whole blood followed by assessment of CD11b activation/expression and cell adhesion under shear-flow. HDL caused a dose-dependent reduction in the activation of CD11b induced by PMA or receptor-dependent agonists. The constituent of HDL responsible for the antiinflammatory effects on CD11b activation was found to be apolipoprotein A-I (apoA-I). Cyclodextrin, but not cyclodextrin/cholesterol complex, also inhibited PMA-induced CD11b activation implicating cholesterol efflux as the main mechanism. This was further confirmed with the demonstration that cholesterol content of lipid rafts diminished after treatment with the cholesterol acceptors. Blocking ABCA1 with an anti-ABCA1 antibody abolished the effect of apoA-I. Furthermore, monocytes derived from a Tangier disease patient definitively confirmed the requirement of ABCA1 in apoA-I-mediated CD11b inhibition. The antiinflammatory effects of apoA-I were also observed in functional models including cell adhesion to an endothelial cell monolayer, monocytic spreading under shear flow, and transmigration. CONCLUSIONS: HDL and apoA-I exhibit an antiinflammatory effect on human monocytes by inhibiting activation of CD11b. ApoA-I acts through ABCA1, whereas HDL may act through several receptors.

Asunto(s)

Apolipoproteína A-I/metabolismo , Aterosclerosis/prevención & control , Inflamación/prevención & control , Lipoproteínas HDL/metabolismo , Monocitos/metabolismo , Transportador 1 de Casete de Unión a ATP , Transportadoras de Casetes de Unión a ATP/metabolismo , Actinas/metabolismo , Aterosclerosis/metabolismo , Aterosclerosis/patología , Antígeno CD11b/metabolismo , Adhesión Celular , Forma de la Célula , Células Cultivadas , Colesterol/deficiencia , Colesterol/metabolismo , Ciclodextrinas/farmacología , Células Endoteliales/metabolismo , Humanos , Inflamación/metabolismo , Inflamación/patología , Microdominios de Membrana/metabolismo , Monocitos/efectos de los fármacos , Monocitos/patología , Adhesividad Plaquetaria , Receptores Depuradores de Clase B , Estrés Mecánico , Enfermedad de Tangier/metabolismo , Acetato de Tetradecanoilforbol/farmacología , Factores de Tiempo

RESUMEN

A pivotal mediator of actin dynamics is the protein cofilin, which promotes filament severing and depolymerization, facilitating the breakdown of existing filaments, and the enhancement of filament growth from newly created barbed ends. It does so in concert with actin interacting protein 1 (Aip1), which serves to accelerate cofilin's activity. While progress has been made in understanding its biochemical functions, the physiologic processes the cofilin/Aip1 complex regulates, particularly in higher organisms, are yet to be determined. We have generated an allelic series for WD40 repeat protein 1 (Wdr1), the mammalian homolog of Aip1, and report that reductions in Wdr1 function produce a dramatic phenotype gradient. While severe loss of function at the Wdr1 locus causes embryonic lethality, macrothrombocytopenia and autoinflammatory disease develop in mice carrying hypomorphic alleles. Macrothrombocytopenia is the result of megakaryocyte maturation defects, which lead to a failure of normal platelet shedding. Autoinflammatory disease, which is bone marrow-derived yet nonlymphoid in origin, is characterized by a massive infiltration of neutrophils into inflammatory lesions. Cytoskeletal responses are impaired in Wdr1 mutant neutrophils. These studies establish an essential requirement for Wdr1 in megakaryocytes and neutrophils, indicating that cofilin-mediated actin dynamics are critically important to the development and function of both cell types.

Asunto(s)

Factores Despolimerizantes de la Actina/metabolismo , Proteínas de Microfilamentos/metabolismo , Mutación/genética , Trombocitopenia/metabolismo , Trombocitopenia/patología , Alelos , Secuencia de Aminoácidos , Animales , Plaquetas/citología , Diferenciación Celular , Movimiento Celular , Secuencia Conservada , Humanos , Inflamación/genética , Inflamación/metabolismo , Inflamación/patología , Megacariocitos/citología , Ratones , Ratones Noqueados , Proteínas de Microfilamentos/química , Proteínas de Microfilamentos/genética , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , Neutrófilos/citología , Neutrófilos/metabolismo , Alineación de Secuencia , Trombocitopenia/genética