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1.
Artigo em Inglês | MEDLINE | ID: mdl-31710506

RESUMO

RATIONALE: Interleukin (IL) 18 is a member of the IL-1 cytokine family and elevated blood IL-18 concentrations associate with disease activity in Macrophage Activation Syndrome (MAS) and poor clinical outcomes in severe inflammatory and septic conditions. OBJECTIVE: While recent investigations provide mechanistic evidence for a contribution of IL-18 to (hyper)inflammation in sepsis and MAS, we sought to study regulatory mechanisms underlying human IL-18 expression. METHODS: Samples from in vivo and in vitro endotoxin re-challenge experiments, inflammatory disease patients and isolated human monocytes treated with various stimulants and drugs were tested for cytokine gene and protein expression. Serum IL-18 expression with or without JAK/STAT-inhibition was analyzed in two MAS mouse models as well as a patient with recurrent MAS. MEASUREMENTS AND MAIN RESULTS: Peripheral blood as well as monocytic IL-18 expression escaped lipopolysacharide (LPS)-induced immunoparalysis. LPS-stimulated primary human monocytes revealed a specific IL-18 expression kinetics controlled by IFNα/ß-signaling. JAK/STAT-inhibition or IFNß-neutralization during LPS-stimulation blunted cytokine expression. Similarly, microtubule destabilizing drugs abrogated LPS-induced IL18 expression, which could be fully reversed by addition of IFNα/ß. Ex vivo analysis of inflammatory disease patients' whole blood revealed strong correlation of type I interferon score and IL18 expression, while JAK/STAT-inhibition in two MAS mouse models as well as a patient with recurrent MAS strongly reduced IL-18 serum levels. CONCLUSION: Our data indicate that IL-18 (but not IL-1ß) production from human monocytes requires cooperate toll-like receptor and IFNα/ß-signaling. Interference with IFNα/ß-expression or signaling following JAK/STAT-inhibition may control catastrophic hyperinflammation in MAS. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/).

2.
Inflamm Bowel Dis ; 2019 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-31560043

RESUMO

BACKGROUND: The inhibition of Janus kinases (JAKs) and subsequent signal transducers and activators of transcription (STATs) by tofacitinib represents a new therapeutic strategy in inflammatory bowel diseases (IBD) as clinical trials have led to approval of tofacitinib for ulcerative colitis (UC) and hint at a possible efficacy for Crohn`s disease (CD). However, the impact of tofacitinib on cellular response of monocytes, which are key players in inflammatory responses, has not been investigated so far. We aimed to analyze JAK/STAT-inhibition by tofacitinib in monocytes of IBD patients and healthy controls. METHODS: Primary monocytes of IBD patients with active disease and healthy controls (n = 18) were analyzed for cytokine expression and phenotype after granulocyte macrophage colony-stimulating factor (GM-CSF)/interferon (IFN)γ-stimulation and tofacitinib pretreatment (1-1000 nM) and capacity to induce Foxp3+-regulatory T cells (Tregs) in cocultures. In total, 20 UC patients and 21 CD patients were included. Additionally, dose-dependent inhibition of JAK/STAT-phosphorylation was analyzed in controls. RESULTS: Pro-inflammatory costimulation with GM-CSF/IFNγ resulted in significant tumor necrosis factor (TNFα) and interleukin (IL)-6 increase, whereas IL-10 expression decreased in monocytes. Tofacitinib modulated the responses of activated monocytes toward a regulatory phenotype through reduced TNFα and IL-6 secretion and enhanced Treg induction in cocultures. However, in monocytes from active IBD patients, higher tofacitinib dosages were needed for blockade of pro-inflammatory cytokines. Tofacitinib induced stronger regulatory phenotypes in monocytes of UC patients, including more effective inhibition of pro-inflammatory pathways and better restoration of anti-inflammatory mechanisms as compared with CD-derived monocytes. CONCLUSION: Tofacitinib dose-dependently reprograms monocytes toward a more regulatory cell type. This beneficial effect possibly results from selective JAK/STAT-blockade by adequate tofacitinib dosage with inhibition of pro-inflammatory responses and permission of a balance-shift toward regulatory pathways.

3.
Ocul Immunol Inflamm ; 27(5): 788-797, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-29771573

RESUMO

Purpose: To characterize peripheral blood T cells in juvenile idiopathic arthritis-associated uveitis (JIAU). Methods: Blood samples were taken from children with JIAU (n = 18), JIA without ocular involvement (n = 11), idiopathic anterior uveitis (IAU, n = 12), and healthy controls (n = 11). Cells were stained for T cell surface markers, and intracellular cytokine staining was performed after cell stimulation and analyzed by flow cytometry. Results: The Th1/Th2 ratio was increased in JIAU patients. Numbers of IL-13-expressing cells an level of IL-13 and IL-10 expression per cell were increased in all patient groups; whereas, percentages of IL-5-expressing T cells were decreased. Numbers of proinflammatory Th17 cells and T cells expressing CTLA-4 were increased in all patient groups; whereas, γ/δ T cell numbers were decreased. Results from JIA and IAU were similar. Conclusion: T cell subtypes and potential T cell function are altered in pediatric patients with uveitis and arthritis as compared to healthy children.


Assuntos
Células Th1/metabolismo , Células Th2/metabolismo , Uveíte Anterior , Adolescente , Artrite Juvenil/complicações , Estudos de Casos e Controles , Criança , Pré-Escolar , Citocinas/metabolismo , Feminino , Humanos , Masculino , Projetos Piloto , Uveíte Anterior/sangue , Uveíte Anterior/imunologia
4.
Ocul Immunol Inflamm ; 27(2): 330-337, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-29020495

RESUMO

PURPOSE: This study analyzed the effect of adalimumab on peripheral blood mononuclear cells (PBMCs) in uveitis. METHODS: PBMCs and serum S100A12 levels from 14 uveitis patients and 28 healthy controls were analyzed. Patient samples were taken before (w0), and 6 (w6) and 12 (w12) weeks after initiation of adalimumab therapy. RESULTS: Monocytes expressing CD124, CD86, CD39, CD115, and MHCII were decreased in patients. Adalimumab induced CD86+ and CD39+ monocytes, and further decreased the frequency of MHCII- and CD124-positive cells. Patients (w0) had increased percentages of Th1-, Th17-, and Th2 cells and T cell subsets showed a pro-inflammatory polarization (p = 0.02 ratio Th17/Treg patients w0 vs controls), which was reduced upon adalimumab treatment (p = 0.05 w0 vs w6). S100A12 levels were increased in patients (p = 0.02) and reduced under treatment (p = 0.02 for w6/w12). CONCLUSIONS: The phenotype of PBMCs from uveitis patients is modified upon adalimumab treatment. Serum S100A12 levels reflect the systemic immune response.


Assuntos
Adalimumab/administração & dosagem , Leucócitos Mononucleares/patologia , Monócitos/patologia , Linfócitos T Reguladores/patologia , Uveíte/tratamento farmacológico , Adolescente , Anti-Inflamatórios/administração & dosagem , Criança , Pré-Escolar , Relação Dose-Resposta a Droga , Feminino , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Masculino , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Estudos Retrospectivos , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/imunologia , Resultado do Tratamento , Uveíte/sangue , Uveíte/imunologia
5.
J Matern Fetal Neonatal Med ; : 1-191, 2018 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-30563403

RESUMO

INTRODUCTION: Sepsis has a grave impact on neonatal morbidity and mortality. Proper timely diagnosis and a subsequently tailored management are crucial to improve neonatal outcome and survival. New diagnostic methods are needed and much effort is directed to this objective. In this work, we aimed to evaluate S100A12 protein as a biomarker of neonatal sepsis. MATERIALS AND METHODS: In this prospective single-center study, 118 preterm and term neonates were enrolled and assigned to four groups: controls, infants with no infection, infants with probable infection and infants with proven infection. Clinical and routine laboratory data, the serum levels of S100A12 and additional cytokines (interleukin (IL)-1ß, IL-2, IL-6, IL-17A, IL-18, IL-22, IL-10 and interferon (IFN)-γ) were assessed. Using stepwise multivariate logistic regression analysis, S100A12 protein was evaluated as a biomarker of neonatal infection. RESULTS: Significant differences of the parameters of complete blood count and level of C-reactive protein were documented between the study/the four groups. The studied marker S100A12 as well as IL-6 and IL-10 were highly significant (p < 0.001) between infected and control groups. S100A12 had a sensitivity of 96.8% and a specificity of 93.3%. Even after adjusting for the confounding factors sex, body weight, gestational age, mode of delivery, number of pregnancies, premature rupture of membranes and preeclampsia S100A12 remained significant between the infected and control groups. CONCLUSIONS: S100A12 may be considered as a new biomarker of neonatal sepsis.

6.
Curr Rheumatol Rep ; 20(9): 53, 2018 07 14.
Artigo em Inglês | MEDLINE | ID: mdl-30008153

RESUMO

PURPOSE OF REVIEW: Current technical advances enable the assessment of the complex changes in body fluid proteomes and thus allow for the discovery of biomarker signatures rather than just following differences of a single marker. In this review, we aim to summarize current approaches to discover and evaluate multi-biomarker panels for improved monitoring of chronic arthritis disease activity. RECENT FINDINGS: Mass spectrometry and affinity proteomic methodologies have been used to identify biomarker panels in synovial fluid, serum, plasma, or urine of pediatric and adult chronic arthritis patients. Notably, despite the numerous efforts to develop new and better biomarker panels, very few have undergone extensive analytical and clinical validation and been adopted into routine use for patient benefit. There remains a significant gap between discovery of chronic arthritis biomarker signatures and their validation for clinical use.


Assuntos
Artrite/diagnóstico , Biomarcadores/análise , Proteômica/métodos , Doença Crônica , Humanos , Espectrometria de Massas/métodos , Reprodutibilidade dos Testes
7.
Clin Immunol ; 190: 84-88, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-28923439

RESUMO

OBJECTIVE: To characterize peripheral blood monocytes in uveitis associated with juvenile idiopathic arthritis (JIAU). METHODS: Peripheral blood monocytes from children with JIA (either with (n = 18) or without uveitis (n = 11)), idiopathic anterior uveitis (IAU; n = 12) and healthy controls (n = 11) were analyzed by flow cytometry. RESULTS: Percentage of CD14 + CD86+ monocytes and CD86 expression on single cell level were significantly higher in all patient groups than in controls, whereas no major differences existed between patient groups. Frequency of CD39+ (p < 0.05 all groups) and CD73+ monocytes (p = 0.03 JIAU vs controls) was elevated in patients. Disease activity did not influence monocyte phenotypes, but in methotrexate-treated JIAU patients numbers of CCR2+ monocytes were reduced and numbers of CD86+ and CD39+ cells increased. CONCLUSION: Children with arthritis or uveitis display a distinct monocytic phenotype when compared to cells from healthy children. Phenotypic changes seem to be neither arthritis- nor uveitis-dependent, but may be modified by treatment.


Assuntos
Artrite Juvenil/imunologia , Monócitos/imunologia , Uveíte Anterior/imunologia , Uveíte/imunologia , Adolescente , Antirreumáticos/uso terapêutico , Artrite Juvenil/sangue , Artrite Juvenil/tratamento farmacológico , Antígeno B7-2/imunologia , Antígeno B7-2/metabolismo , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Receptores de Lipopolissacarídeos/imunologia , Receptores de Lipopolissacarídeos/metabolismo , Masculino , Metotrexato/uso terapêutico , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Resultado do Tratamento , Uveíte/sangue , Uveíte/tratamento farmacológico , Uveíte Anterior/sangue , Uveíte Anterior/tratamento farmacológico
8.
J Am Soc Nephrol ; 29(3): 1011-1019, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29196304

RESUMO

Whether earlier initiation of RRT in critically ill patients with AKI can improve outcomes remains debated. We examined follow-up data from a large clinical trial to prospectively investigate the long-term outcomes associated with the timing of RRT initiation in such patients. We extended the follow-up of patients in the Early Versus Delayed Initiation of RRT in Critically Ill Patients with AKI (ELAIN) Trial from 90 days to 1 year after randomization for 230 (99.6%) patients. The primary outcome was a composite of major adverse kidney events (persistent renal dysfunction, dialysis dependence, and mortality) at 1 year. Secondary outcomes included inflammatory markers. Overall, 72 of 111 (64.9%) and 106 of 119 (89.1%) patients met the primary outcome in the early (stage 2 AKI) and delayed (stage 3 AKI) initiation groups, respectively (odds ratio [OR] with early initiation, 0.23; 95% confidence interval [95% CI], 0.11 to 0.45; P< 0.001). The early initiation group had a 1-year all-cause mortality rate (56 of 111 [50.2%]) significantly lower than that of the delayed initiation group (83 of 119 [69.8%]; absolute difference, -19.6%; 95% CI, -32.0% to -7.2%; P<0.01). After 1 year, 16 of 55 (29.1%) and 23 of 36 (63.9%) surviving patients in the early and delayed groups, respectively, failed to recover renal function (absolute difference, -34.8%; 95% CI, -54.6% to -15.0%; P=0.001). In conclusion, early initiation of RRT in these critically ill patients with AKI significantly reduced the occurrence of major adverse kidney events, reduced mortality, and enhanced renal recovery at 1 year.


Assuntos
Lesão Renal Aguda/terapia , Interleucinas/sangue , Oxirredutases Intramoleculares/sangue , Fatores Inibidores da Migração de Macrófagos/sangue , Insuficiência Renal Crônica/etiologia , Insuficiência Renal Crônica/terapia , Tempo para o Tratamento , Lesão Renal Aguda/complicações , Lesão Renal Aguda/fisiopatologia , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Estado Terminal , Seguimentos , Humanos , Inflamação/sangue , Interleucina-10/sangue , Interleucina-18/sangue , Interleucina-6/sangue , Interleucina-8/sangue , Mortalidade , Recuperação de Função Fisiológica , Insuficiência Renal Crônica/fisiopatologia , Terapia de Substituição Renal , Fatores de Tempo
9.
Arthritis Rheumatol ; 69(7): 1480-1494, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28296284

RESUMO

OBJECTIVE: Systemic-onset juvenile idiopathic arthritis (JIA) is speculated to follow a biphasic course, with an initial systemic disease phase driven by innate immune mechanisms and interleukin-1ß (IL-1ß) as a key cytokine and a second chronic arthritic phase that may be dominated by adaptive immunity and cytokines such as IL-17A. Although a recent mouse model points to a critical role of IL-17-expressing γ/δ T cells in disease pathology, in humans, both the prevalence of IL-17 and the role of IL-17-producing cells are still unclear. METHODS: Serum samples from systemic JIA patients and healthy pediatric controls were analyzed for the levels of IL-17A and related cytokines. Whole blood samples were studied for cellular expression of IL-17 and interferon-γ (IFNγ). CD4+ and γ/δ T cells isolated from the patients and controls were assayed for cytokine secretion in different culture systems. RESULTS: IL-17A was prevalent in sera from patients with active systemic JIA, while both ex vivo and in vitro experiments revealed that γ/δ T cells overexpressed this cytokine. This was not seen with CD4+ T cells, which expressed strikingly low levels of IFNγ. Therapeutic IL-1 blockade was associated with partial normalization of both cytokine expression phenotypes. Furthermore, culturing healthy donor γ/δ T cells in serum from systemic JIA patients or in medium spiked with IL-1ß, IL-18, and S100A12 induced IL-17 overexpression at levels similar to those observed in the patients' cells. CONCLUSION: A systemic JIA cytokine environment may prime γ/δ T cells in particular for IL-17A overexpression. Thus, our observations in systemic JIA patients strongly support a pathophysiologic role of these cells, as proposed by the recent murine model.


Assuntos
Artrite Juvenil/imunologia , Interleucina-17/imunologia , Linfócitos T/imunologia , Imunidade Adaptativa/imunologia , Adolescente , Adulto , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados , Antirreumáticos/uso terapêutico , Artrite Juvenil/tratamento farmacológico , Linfócitos T CD4-Positivos/imunologia , Estudos de Casos e Controles , Quimiocina CXCL10/imunologia , Quimiocina CXCL9/imunologia , Criança , Pré-Escolar , Citocinas/imunologia , Feminino , Humanos , Imunidade Inata/imunologia , Interferon gama/imunologia , Proteína Antagonista do Receptor de Interleucina 1/uso terapêutico , Interleucina-18/imunologia , Interleucina-18/farmacologia , Interleucina-1beta/imunologia , Interleucina-1beta/farmacologia , Subunidade p19 da Interleucina-23/imunologia , Interleucina-6/imunologia , Masculino , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Proteína S100A12/imunologia , Proteína S100A12/farmacologia , Linfócitos T/metabolismo , Adulto Jovem
10.
Clin Immunol ; 2016 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-27989897

RESUMO

We analyzed phenotype and function of peripheral blood mononuclear cells in 9 patients with active idiopathic intermediate uveitis (IIU) before and after 6 and 12weeks of systemic corticosteroid (CS) treatment and compared to 28 healthy individuals. Monocytes from IIU patients showed increased MHCII expression compared with controls (p=0.09). Treatment reduced expression of MHCII, CD86, CD39 and CD124 (all p<0.05), whereas the percentage of CD121b-expressing monocytes was increased by week 6 (p=0.039). Patients showed alterations in T cell polarization (Th1/Th2 ratio: patients 5.2 versus controls 3.1, p=0.054; Th17/Treg ratio: 3.0 versus 1.7, p=0.027). S100A12 serum levels were higher in active IIU (p=0.057). Phagocytosis, oxidative burst and serum cytokine levels did not differ between patients and controls, and were not altered by treatment. In conclusion, monocytes from patients with active IIU show increased co-stimulatory capacities, which are modulated by systemic CS treatment, whereas innate immune cell functions are not altered.

11.
Arthritis Rheumatol ; 68(12): 3010-3022, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27333294

RESUMO

OBJECTIVE: Familial Mediterranean fever (FMF) is an autoinflammatory disorder caused by pyrin-encoding MEFV mutations. Patients present with recurrent but self-limiting episodes of acute inflammation and often have persistent subclinical inflammation. The pathophysiology is only partially understood, but neutrophil overactivation is a hallmark of the disease. S100A12 is a neutrophil-derived proinflammatory danger signal that is strongly elevated in active FMF. This study was undertaken to characterize the secretory activity of neutrophils in vitro and investigate the association of S100A12 with disease activity and genotype in patients with FMF. METHODS: Neutrophils from FMF patients carrying the p.M694V mutation (1 compound heterozygous and 5 homozygous) and neutrophils from 4 healthy control subjects were purified and stimulated in vitro. Neutrophil secretion of S100A12, interleukin-18 (IL-18), IL-1ß, and caspase 1 was determined. Based on these in vitro analyses, serum concentrations of S100A12, IL-18, and IL-1ß were also analyzed in 128 clinically and genetically characterized patients with FMF. RESULTS: In vitro, unstimulated neutrophils from p.M694V-positive patients spontaneously secreted more S100A12, IL-18, and caspase 1 compared to neutrophils from healthy controls. Serum concentrations of S100A12 correlated with disease activity and genotype, with the levels being highest in homozygous patients and with compound heterozygotes displaying higher levels than heterozygotes. Compared to individuals negative for the p.M694V mutation, heterozygous, compound heterozygous, or homozygous p.M694V-positive patients had higher serum levels of S100A12 and IL-18 during inactive and subclinical disease. CONCLUSION: The FMF phenotype is known to be more severe in patients carrying the p.M694V mutation. This report describes 2 molecules secreted by unconventional secretory pathways, S100A12 and IL-18, whose concentrations correlated with clinical disease activity and genotype in patients with FMF. In this clinically and genetically heterogeneous disease, management of these surrogate markers might help to improve patient care and outcomes.


Assuntos
Caspase 1/metabolismo , Febre Familiar do Mediterrâneo/genética , Interleucina-18/metabolismo , Interleucina-1beta/metabolismo , Neutrófilos/metabolismo , Pirina/genética , Proteína S100A12/metabolismo , Adolescente , Adulto , Estudos de Casos e Controles , Caspase 1/sangue , Criança , Pré-Escolar , Febre Familiar do Mediterrâneo/sangue , Febre Familiar do Mediterrâneo/imunologia , Feminino , Genótipo , Heterozigoto , Homozigoto , Humanos , Técnicas In Vitro , Interleucina-18/sangue , Interleucina-1beta/sangue , Masculino , Pessoa de Meia-Idade , Neutrófilos/imunologia , Proteína S100A12/sangue , Adulto Jovem
12.
Eur J Immunol ; 46(8): 1997-2007, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27159026

RESUMO

In patients with juvenile idiopathic arthritis (JIA), increased release of IFN-γ and GM-CSF in cells infiltrating synovial tissue can be a potent driver of monocyte activation. Given the fundamental role of monocyte activation in remodeling the early phases of inflammatory responses, here we analyze the GM-CSF/IFN-γ induced activity of human monocytes in such a situation in vitro and in vivo. Monocytes from healthy donors were isolated and stimulated with GM-CSF ± IFN-γ. Monocyte activation and death were analyzed by flow cytometry, immunofluorescence microscopy, ELISA, and qPCR. T-cell GM-CSF/IFN-γ expression and monocyte function were determined in synovial fluid and peripheral blood from 15 patients with active JIA and 21 healthy controls. Simultaneous treatment with GM-CSF and IFN-γ induces cell death of monocytes. This cell death is partly cathepsin B-associated and has morphological characteristics of necrosis. Monocytes responding to costimulation with strong proinflammatory activities are consequently eliminated. Monocytes surviving this form of hyperactivation retain normal cytokine production. Cathepsin B activity is increased in monocytes isolated from synovial fluid from patients with active arthritis. Our data suggest GM-CSF/IFN-γ induced cell death of monocytes as a novel mechanism to eliminate overactivated monocytes, thereby potentially balancing inflammation and autoimmunity in JIA.


Assuntos
Artrite Juvenil/imunologia , Morte Celular , Citocinas/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Interferon gama/farmacologia , Monócitos/imunologia , Adolescente , Autoimunidade , Estudos de Casos e Controles , Catepsina B/metabolismo , Feminino , Citometria de Fluxo , Humanos , Ativação Linfocitária , Masculino , Líquido Sinovial
13.
J Immunol ; 194(5): 2424-38, 2015 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-25653427

RESUMO

Human and murine studies showed that GM-CSF exerts beneficial effects in intestinal inflammation. To explore whether GM-CSF mediates its effects via monocytes, we analyzed effects of GM-CSF on monocytes in vitro and assessed the immunomodulatory potential of GM-CSF-activated monocytes (GMaMs) in vivo. We used microarray technology and functional assays to characterize GMaMs in vitro and used a mouse model of colitis to study GMaM functions in vivo. GM-CSF activates monocytes to increase adherence, migration, chemotaxis, and oxidative burst in vitro, and primes monocyte response to secondary microbial stimuli. In addition, GMaMs accelerate epithelial healing in vitro. Most important, in a mouse model of experimental T cell-induced colitis, GMaMs show therapeutic activity and protect mice from colitis. This is accompanied by increased production of IL-4, IL-10, and IL-13, and decreased production of IFN-γ in lamina propria mononuclear cells in vivo. Confirming this finding, GMaMs attract T cells and shape their differentiation toward Th2 by upregulating IL-4, IL-10, and IL-13 in T cells in vitro. Beneficial effects of GM-CSF in Crohn's disease may possibly be mediated through reprogramming of monocytes to simultaneously improved bacterial clearance and induction of wound healing, as well as regulation of adaptive immunity to limit excessive inflammation.


Assuntos
Imunidade Adaptativa/efeitos dos fármacos , Colite/tratamento farmacológico , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Intestino Grosso/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Transferência Adotiva , Animais , Adesão Celular/efeitos dos fármacos , Quimiotaxia/efeitos dos fármacos , Colite/imunologia , Colite/patologia , Regulação da Expressão Gênica , Humanos , Interferon gama/farmacologia , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-13/genética , Interleucina-13/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Interleucina-4/farmacologia , Intestino Grosso/imunologia , Intestino Grosso/patologia , Camundongos , Camundongos Knockout , Monócitos/citologia , Monócitos/imunologia , Cultura Primária de Células , Explosão Respiratória/efeitos dos fármacos , Fatores de Transcrição SOXF/deficiência , Fatores de Transcrição SOXF/genética , Fatores de Transcrição SOXF/imunologia , Transdução de Sinais , Linfócitos T/imunologia , Linfócitos T/patologia , Linfócitos T/transplante
14.
Cell Mol Gastroenterol Hepatol ; 1(4): 433-449.e1, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28210690

RESUMO

BACKGROUND & AIMS: Granulocyte macrophage colony-stimulating factor (GM-CSF) treatment induces clinical response in patients with active Crohn's disease. To explore whether monocytes mediate GM-CSF effects in vivo, we used a mouse model of chronic colitis induced by dextran sulfate sodium (DSS). METHODS: Murine bone marrow-derived monocytes were activated with GM-CSF in vitro, and gene expression, phenotype, and function of GM-CSF-activated monocytes (GMaM) were analyzed. Therapeutic effects of GMaM were assessed in a model of chronic colitis induced by repeated cycles of DSS. Monocytes were administered intravenously and their immunomodulatory functions were evaluated in vivo by clinical monitoring, histology, endoscopy, immunohistochemistry, and expression of inflammatory markers in the colon. The distribution of injected monocytes in the intestine was measured by in vivo imaging. RESULTS: GMaM expressed significantly higher levels of anti-inflammatory molecules. Production of reactive oxygen species was also increased while phagocytosis and adherence were decreased. GMaM up-regulated CD39 and CD73, which allows the conversion of adenosine triphosphate into adenosine and coincided with the induction of Foxp3+ (forkhead-box-protein P3 positive) regulatory T cells (Treg) in cocultures of GMaM and naive T cells. In chronic DSS-induced colitis, adoptive transfer of GMaM led to significant clinical improvement, as demonstrated by reduced weight loss, inflammatory infiltration, ulceration, and colon shrinkage. As GMaM migrated faster and persisted longer in the inflamed intestine compared with control monocytes, their presence induced Treg generation in vivo. CONCLUSIONS: GM-CSF leads to specific monocyte activation that modulates experimental colitis via mechanisms that include the induction of Treg. We demonstrate a possible mechanism of Treg induction through CD39 and CD73 expression on monocytes.

15.
Immunol Res ; 61(1-2): 4-10, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25407645

RESUMO

Histologic chorioamnionitis (HCA) is an intrauterine status of inflammation which may lead to the fetal inflammatory response syndrome. Inflammation is a pathogenetic mechanism also of preeclampsia, although not of microbial origin. The aim of the present pilot study was to evaluate the pattern of inflammatory cytokines in mothers and high-risk preterm infants during the perinatal period. Concentrations of proinflammatory and anti-inflammatory cytokines and C-reactive protein were evaluated in maternal, cord, and neonatal blood of very preterm infants <1,500 g birth weight. Histologic examinations of placentae and umbilical cords were performed. The 65 mother-neonate pairs enrolled were subdivided into three groups: (1) HCA group (n = 15), (2) preeclampsia group (n = 17), and (3) control group, in the absence of HCA/preeclampsia (n = 33). Maternal Interleukin (IL)-6 levels were significantly higher in women of the HCA group compared with the preeclampsia and control groups (p < 0.05). IL-22 was detected in nearly all maternal samples [median value 693.115 pg/ml (599.91-809.91 pg/ml)], with no statistical difference between the groups, but with a tendency to increased levels among preeclamptic women. Increased concentrations of IL-22 were detected in cord blood of neonates exposed to preeclampsia, compared with controls and infants exposed to HCA (p < 0.05). We speculate that the tendentially higher concentrations of IL-22 in preeclamptic mothers and the significantly higher concentrations in cord blood may reflect placental dysfunction and the underlying reparative processes at the maternal-fetal interface. Therefore, IL-22 could be an important biomarker of inflammation in preeclampsia.


Assuntos
Inflamação/metabolismo , Interleucinas/metabolismo , Adulto , Biomarcadores/sangue , Biomarcadores/metabolismo , Proteína C-Reativa/metabolismo , Corioamnionite/sangue , Corioamnionite/metabolismo , Citocinas/sangue , Citocinas/metabolismo , Feminino , Sangue Fetal , Humanos , Recém-Nascido , Inflamação/sangue , Inflamação/diagnóstico , Interleucinas/sangue , Contagem de Leucócitos , Masculino , Projetos Piloto , Placenta/metabolismo , Placenta/patologia , Pré-Eclâmpsia/sangue , Pré-Eclâmpsia/metabolismo , Gravidez
16.
J Immunol ; 193(3): 1090-9, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-24990080

RESUMO

Glucocorticoids (GCs) are used as first-line therapies for generalized suppression of inflammation (e.g., allergies or autoimmune diseases), but their long-term use is limited by severe side effects. Our previous work revealed that GCs induced a stable anti-inflammatory phenotype in monocytes, the GC-stimulated monocytes (GCsMs) that we exploited for targeted GC-mediated therapeutic effects. We demonstrate that GCsMs interact with T cells in suppressing proliferation, as well as cytokine release of CD8(+) and, especially, CD4(+) T cells in vitro, and that they support generation of Foxp3(+) cells. Therefore, we tested their immunosuppressive potential in CD4(+) T cell-induced colitis in vivo. We found that injection of GCsMs into mice with severe colitis abolished the inflammation and resulted in significant clinical improvement within a few days. T cells recovered from GCsM-treated mice exhibited reduced secretion of proinflammatory cytokines IFN-γ and IL-17. Furthermore, clusters of Foxp3(+) CD4(+) T cells were detectable at local sites of inflammation in the colon. Thus, GCsMs are able to modify T cell responses in vitro and in vivo, as well as to downregulate and clinically cure severe T cell-mediated colitis.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/transplante , Comunicação Celular/imunologia , Glucocorticoides/farmacologia , Tolerância Imunológica/imunologia , Mediadores da Inflamação/administração & dosagem , Monócitos/imunologia , Animais , Anticorpos Neutralizantes/fisiologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Comunicação Celular/efeitos dos fármacos , Técnicas de Cocultura , Colite/tratamento farmacológico , Colite/imunologia , Colite/patologia , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/imunologia , Glucocorticoides/efeitos adversos , Tolerância Imunológica/efeitos dos fármacos , Mediadores da Inflamação/efeitos adversos , Interleucina-10/antagonistas & inibidores , Interleucina-10/biossíntese , Interleucina-10/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Monócitos/efeitos dos fármacos , Monócitos/patologia
17.
Endocrinology ; 155(10): 3899-908, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25057793

RESUMO

Glucocorticoids (GCs) constitute a highly pleiotropic class of drugs predominantly employed in the treatment of inflammatory diseases. In our search for new mechanisms of action, we identified a hitherto unknown effect of GCs in the gastrointestinal tract. We found that oral administration of dexamethasone (Dex) to mice caused an enlargement of the stomach due to the induction of gastroparesis and that this effect was abolished in GR(dim) mice carrying the A458T mutation in the GC receptor (GR). Gastroparesis was unrelated to the enhanced gastric acid secretion observed after Dex treatment, although both effects were mediated by the same molecular mechanism of the GR. Using conditional GR-knockout mice, we could further rule out that GC effects on enterocytes or myeloid cells were involved in the induction of gastroparesis. In contrast, we found that Dex upregulated arginase 2 (Arg2) in the stomach both at the mRNA and protein level. This suggests that GC treatment leads to a depletion of l-arginine thereby impeding the production of nitric oxide (NO), which is required for gastric motility. We tested this hypothesis by supplementing the drinking water of the mice with exogenous l-arginine to compensate for the presumed shortage of this major substrate of NO synthases. Importantly, this measure completely prevented both the enlargement of the stomach and the induction of gastroparesis after Dex treatment. Our findings raise considerations of combining orally applied GCs with l-arginine to improve tolerability of GC treatment and provide a possible explanation for the antiemetic effects of GCs widely exploited in chemotherapy.


Assuntos
Arginina/deficiência , Dexametasona/efeitos adversos , Gastroparesia/induzido quimicamente , Glucocorticoides/efeitos adversos , Animais , Arginase/genética , Arginase/metabolismo , Dexametasona/administração & dosagem , Feminino , Gastroparesia/genética , Gastroparesia/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Glucocorticoides/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/metabolismo , Regulação para Cima/efeitos dos fármacos
18.
Arthritis Rheumatol ; 66(5): 1327-39, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24470119

RESUMO

OBJECTIVE: Monosodium urate monohydrate (MSU) crystal-induced interleukin-1ß (IL-1ß) secretion is a critical factor in the pathogenesis of gout. However, without costimulation by a proIL-1ß-inducing factor, MSU crystals alone are insufficient to induce IL-1ß secretion. The responsible costimulatory factors that act as a priming endogenous signal in vivo are not yet known. We undertook this study to analyze the costimulatory properties of myeloid-related protein 8 (MRP-8) and MRP-14 (endogenous Toll-like receptor 4 [TLR-4] agonists) in MSU crystal-induced IL-1ß secretion and their relevance in gout. METHODS: MRP-8/MRP-14 was measured in paired serum and synovial fluid samples by enzyme-linked immunosorbent assay (ELISA) and localized in synovial tissue from gout patients by immunohistochemistry. Serum levels were correlated with disease activity, and MSU crystal-induced release of MRPs from human phagocytes was measured. Costimulatory effects of MRP-8 and MRP-14 on MSU crystal-induced IL-1ß secretion from phagocytes were analyzed in vitro by ELISA, Western blotting, and polymerase chain reaction. The impact of MRP was tested in vivo in a murine MSU crystal-induced peritonitis model. RESULTS: MRP-8/MRP-14 levels were elevated in the synovium, tophi, and serum of patients with gout and correlated with disease activity. MRP-8/MRP-14 was released by MSU crystal-activated phagocytes and increased MSU crystal-induced IL-1ß secretion in a TLR-4-dependent manner. Targeted deletion of MRP-14 in mice led to a moderately reduced response of MSU crystal-induced inflammation in vivo. CONCLUSION: MRP-8 and MRP-14, which are highly expressed in gout, are enhancers of MSU crystal-induced IL-1ß secretion in vitro and in vivo. These endogenous TLR-4 ligands released by activated phagocytes contribute to the maintenance of inflammation in gout.


Assuntos
Calgranulina A/metabolismo , Calgranulina B/metabolismo , Gota/metabolismo , Inflamação/metabolismo , Ácido Úrico/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Células Cultivadas , Cristalização , Modelos Animais de Doenças , Feminino , Humanos , Interleucina-1beta/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pessoa de Meia-Idade , Fagócitos/metabolismo , Receptor 4 Toll-Like/deficiência , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
19.
PLoS One ; 8(4): e62761, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23658649

RESUMO

Recent developments suggest a causal link between inflammation and impaired bacterial clearance in Crohn's disease (CD) due to alterations of intestinal macrophages. Studies suggest that excessive inflammation is the consequence of an underlying immunodeficiency rather than the primary cause of CD pathogenesis. We characterized phenotypic and functional features of peripheral blood monocytes of patients with quiescent CD (n = 18) and healthy controls (n = 19) by analyses of cell surface molecule expression, cell adherence, migration, chemotaxis, phagocytosis, oxidative burst, and cytokine expression and secretion with or without lipopolysaccharide (LPS) priming. Peripheral blood monocytes of patients with inactive CD showed normal expression of cell surface molecules (CD14, CD16, CD116), adherence to plastic surfaces, spontaneous migration, chemotaxis towards LTB4, phagocytosis of E. coli, and production of reactive oxygen species. Interestingly, peripheral blood monocytes of CD patients secreted higher levels of IL1ß (p<.05). Upon LPS priming we found a decreased release of IL10 (p<.05) and higher levels of CCL2 (p<.001) and CCL5 (p<.05). The expression and release of TNFα, IFNγ, IL4, IL6, IL8, IL13, IL17, CXCL9, and CXCL10 were not altered compared to healthy controls. Based on our phenotypic and functional studies, peripheral blood monocytes from CD patients in clinical remission were not impaired compared to healthy controls. Our results highlight that defective innate immune mechanisms in CD seems to play a role in the (inflamed) intestinal mucosa rather than in peripheral blood.


Assuntos
Doença de Crohn/imunologia , Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Monócitos/efeitos dos fármacos , Adulto , Antígenos CD/genética , Antígenos CD/imunologia , Estudos de Casos e Controles , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Quimiotaxia/efeitos dos fármacos , Doença de Crohn/genética , Doença de Crohn/patologia , Escherichia coli/imunologia , Feminino , Expressão Gênica/imunologia , Humanos , Interferon gama/genética , Interferon gama/imunologia , Interleucinas/genética , Interleucinas/imunologia , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Monócitos/patologia , Fagocitose/efeitos dos fármacos , Indução de Remissão , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
20.
Am J Respir Crit Care Med ; 187(12): 1324-34, 2013 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-23611140

RESUMO

RATIONALE: S100A12 is overexpressed during inflammation and is a marker of inflammatory disease. Furthermore, it has been ascribed to the group of damage-associated molecular pattern molecules that promote inflammation. However, the exact role of human S100A12 during early steps of immune activation and sepsis is only partially described thus far. OBJECTIVES: We analyzed the activation of human monocytes by granulocyte-derived S100A12 as a key function of early inflammatory processes and the development of sepsis. METHODS: Circulating S100A12 was determined in patients with sepsis and in healthy subjects with experimental endotoxemia. The release of human S100A12 from granulocytes as well as the promotion of inflammation by activation of human monocytes after specific receptor interaction was investigated by a series of in vitro experiments. MEASUREMENTS AND MAIN RESULTS: S100A12 rises during sepsis, and its expression and release from granulocytes is rapidly induced in vitro and in vivo by inflammatory challenge. A global gene expression analysis of S100A12-activated monocytes revealed that human S100A12 induces inflammatory gene expression. These effects are triggered by an interaction of S100A12 with Toll-like receptor 4 (TLR4). Blocking S100A12 binding to TLR4 on monocytes or TLR4 expressing cell lines (HEK-TCM) abrogates the respective inflammatory signal. On the contrary, blocking S100A12 binding to its second proposed receptor (receptor for advanced glycation end products [RAGE]) has no significant effect on inflammatory signaling in monocytes and RAGE-expressing HEK293 cells. CONCLUSIONS: Human S100A12 is an endogenous TLR4 ligand that induces monocyte activation, thereby acting as an amplifier of innate immunity during early inflammation and the development of sepsis.


Assuntos
Inflamação/etiologia , Monócitos/fisiologia , Proteínas S100/fisiologia , Sepse/imunologia , Receptor 4 Toll-Like/fisiologia , Adulto , Idoso , Células Cultivadas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas S100/sangue , Proteína S100A12 , Sepse/sangue , Receptor 4 Toll-Like/sangue , Adulto Jovem
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