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Leucemia Mielomonocítica Crónica , Monocitos , Síndromes Mielodisplásicos , Femenino , Proteínas Ligadas a GPI/sangre , Proteínas Ligadas a GPI/inmunología , Humanos , Leucemia Mielomonocítica Crónica/sangre , Leucemia Mielomonocítica Crónica/inmunología , Leucemia Mielomonocítica Crónica/patología , Receptores de Lipopolisacáridos/sangre , Receptores de Lipopolisacáridos/inmunología , Masculino , Monocitos/inmunología , Monocitos/metabolismo , Monocitos/patología , Monocinas/sangre , Monocinas/inmunología , Síndromes Mielodisplásicos/sangre , Síndromes Mielodisplásicos/inmunología , Síndromes Mielodisplásicos/patología , Receptores de IgG/sangre , Receptores de IgG/inmunologíaAsunto(s)
Monocinas/sangre , Micosis Fungoide/sangre , Rayos Ultravioleta , Terapia Ultravioleta , Adulto , Femenino , Humanos , MasculinoRESUMEN
BACKGROUND: The ability to measure T-cell responses to antigens is proving critical in the field of vaccine development and for understanding immunity to pathogens, allergens and self-antigens. Although a variety of technologies exist for this purpose IFNγ-ELISpot assays are widely used because of their sensitivity and simplicity. However, ELISpots cannot be performed on whole blood, and require relatively large volumes of blood to yield sufficient numbers of peripheral blood mononuclear cells. To address these deficiencies, we describe an assay that measures antigen-specific T cell responses through changes in monokine gene transcription. The biological amplification of the IFNγ signal generated by this assay provides sensitivity comparable to ELISpot, but with the advantage that responses can be quantified using small volumes of whole blood. METHODS: Whole blood or peripheral blood mononuclear cells (PBMCs) from healthy controls and immunosuppressed recipients of solid organ transplants were incubated with peptide pools covering viral and control antigens or mitogen for 20 hours. Total RNA was extracted and reverse transcribed before amplification in a TaqMan qPCR reaction using primers and probes specific for MIG (CXCL9), IP-10 (CXCL10) and HPRT. The induction of MIG and IP-10 in response to stimuli was analysed and the results were compared with those obtained by ELISpot. RESULTS: Antigen-specific T cell responses can be measured through the induction of MIG or IP-10 gene expression in PBMCs or whole blood with results comparable to those achieved in ELISpot assays. The biological amplification generated by IFNγ-R signaling allows responses to be detected in as little as 25 uL of whole blood and enables the assay to retain sensitivity despite storage of samples for up to 48 hours prior to processing. CONCLUSIONS: A monokine-based reporter assay provides a sensitive measure of antigen-specific T cell activation. Assays can be performed on small volumes of whole blood and remain accurate despite delays in processing. This assay may be a useful tool for studying T cell responses, particularly when samples are limited in quantity or when storage or transportation are required before processing.
Asunto(s)
Ensayo de Immunospot Ligado a Enzimas/métodos , Monocinas/sangre , Especificidad del Receptor de Antígeno de Linfocitos T/inmunología , Linfocitos T/inmunología , Células Cultivadas , Humanos , Terapia de Inmunosupresión , Interferón gamma/farmacología , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Recuento de Linfocitos , Receptores de Citocinas/genética , Receptores de Citocinas/metabolismo , Proteínas Recombinantes/farmacología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Linfocitos T/efectos de los fármacosRESUMEN
BACKGROUND: The smallpox vaccine is associated with more serious adverse events than any other live attenuated vaccine in use today. Although studies have examined serum cytokine levels in primary vaccine recipients at 1 and 3-5 weeks after vaccination with the smallpox vaccine, serial measurements have not been performed, and studies in revaccinated subjects have not been conducted. METHODS: We analyzed cytokine responses in both primary vaccine recipients and revaccinated subjects every other day for 2 weeks after vaccination. RESULTS: Primary vaccine recipients had maximal levels of granulocyte-colony-stimulating factor on days 6-7 after vaccination; peak levels of tumor necrosis factor (TNF)-alpha, soluble TNF receptor 1, interferon (IFN)-gamma, IFN-inducible protein-10 (IP-10), interleukin (IL)-6, and tissue inhibitor of metalloproteinases-1 on days 8-9 after vaccination; peak levels of soluble TNF receptor 2 and monokine induced by IFN-gamma (MIG) on days 10-11 after vaccination; and peak levels of granulocyte-macrophage-colony-stimulating factor on days 12-13 after vaccination. Primary vaccine recipients were significantly more likely to have higher peak levels of IFN-gamma, IP-10, and MIG after vaccination than were revaccinated subjects. Primary vaccine recipients were significantly more likely to have fatigue, lymphadenopathy, and headache, as well as a longer duration of these symptoms and more hours missed from work, compared with revaccinated subjects. CONCLUSIONS: The increased frequency and duration of symptoms observed in primary vaccine recipients, compared with revaccinated subjects, paralleled the increases in serum cytokine levels in these individuals. TRIAL REGISTRATION: Clinicaltrials.gov identifier NCT00325975.
Asunto(s)
Citocinas/sangre , Vacuna contra Viruela/farmacología , Adulto , Quimiocina CXCL10/sangre , Femenino , Factor Estimulante de Colonias de Granulocitos/sangre , Humanos , Interferón gamma/sangre , Interferones/sangre , Interleucinas/sangre , Cinética , Masculino , Persona de Mediana Edad , Monocinas/sangre , Receptores del Factor de Necrosis Tumoral/sangre , Vacuna contra Viruela/administración & dosificación , Vacuna contra Viruela/efectos adversos , Vacuna contra Viruela/inmunología , Factor de Necrosis Tumoral alfa/sangre , Adulto JovenRESUMEN
AIMS/INTRODUCTION: Our aims were to examine the add-on effects of a sodium-glucose cotransporter 2 inhibitor, dapagliflozin, compared with existing antidiabetes treatments, on anthropometric/metabolic parameters, the levels of an endocrine regulator, fibroblast growth factor 21 (FGF21); a skeletal muscle mass (SMM) negative regulator, myostatin; and a metabolic regulator, irisin, in patients with type 2 diabetes. MATERIALS AND METHODS: A total of 54 patients with type 2 diabetes were randomly divided into dapagliflozin and control groups. The dapagliflozin group received dapagliflozin 5 mg/day in addition to conventional therapy for 24 weeks. The primary outcome was the change in the level of serum FGF21 from baseline. The secondary outcomes included changes from baseline in anthropometric/metabolic parameters and serum levels of myostatin and irisin. RESULTS: Bodyweight decreased in the dapagliflozin group compared with the control group (P < 0.001), but the changes in SMM were not significant between the groups (P = 0.611), thereby elevating the ratio of SMM-to-bodyweight in the dapagliflozin group (P = 0.028). Myostatin levels were significantly decreased (P = 0.010), and irisin levels showed a nearly significant reduction (P = 0.052) in the dapagliflozin group compared with the control group, whereas FGF21 levels did not change significantly from baseline to the end of the intervention in both the dapagliflozin (P = 0.673) and the control (P = 0.823) groups. CONCLUSIONS: Dapagliflozin add-on therapy in patients with type 2 diabetes reduced myostatin levels significantly and maintained SMM, without significant changes in FGF21 levels.
Asunto(s)
Compuestos de Bencidrilo/uso terapéutico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Factores de Crecimiento de Fibroblastos/sangre , Glucósidos/uso terapéutico , Monocinas/sangre , Músculo Esquelético/patología , Inhibidores del Cotransportador de Sodio-Glucosa 2/uso terapéutico , Anciano , Pueblo Asiatico , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/patología , Femenino , Humanos , Japón , Masculino , Persona de Mediana Edad , Resultado del TratamientoRESUMEN
Interactions between cytokines play an important role in the development of thyroid autoimmunity. Using enzyme-linked immunosorbent assay we investigated serum concentrations of soluble interleukin-2 receptor (sIL-2R), interferon-gamma, tumour necrosis factor (TNF)-alpha, interleukin (IL)-10, CD30, monokine induced by interferon-gamma (MIG), cytotoxic T lymphocyte antigen-4 and markers of apoptosis decoy receptor 3 and Bcl-2 in 28 patients with hyperthyroid Graves' disease (GD), 24 patients with untreated Hashimoto's thyroiditis (HT) and 15 healthy controls. TNF-alpha, IL-10 and sIL-2R were higher in GD compared with HT and controls (TNF-alpha: 8.79 in GD versus 2.54 pg/ml in HT, P = 0.01; IL-10: 10.00 versus 3.10 versus 3.10 pg/ml, P(1) < 0.001, P(2) = 0.005; sIL-2R: 1.26 versus 0.64 versus 0.46 ng/ml, P < 0.001). MIG and CD30 were higher in HT compared with controls (649.22 +/- 262.55 versus 312.95 +/- 143.35 pg/ml, P = 0.037, 6.57 +/- 2.35 versus 3.03 +/- 1.04 U/ml, P = 0.036 respectively). In GD sIL-2R decreased when the euthyroid state was achieved (1.31 +/- 0.64 versus 0.260 +/- 0.11, n = 12, P < 0.001). sIL-2R correlated positively with free thyroxine (FT4) (R = 0.521, P = 0.000) and negatively with thyroid stimulating hormone (TSH) (R = -0.472, P = 0.00132). MIG correlated negatively with FT4 (R = -0.573, P = 0.00234) and positively with TSH (R = 0.462, P = 0.0179). The results suggest that serum concentrations of sIL-2R and MIG are related to thyroid function rather than to activation of autoimmunity.
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Interferón gamma/inmunología , Monocinas/sangre , Receptores de Interleucina-2/inmunología , Glándula Tiroides/metabolismo , Tiroiditis Autoinmune/inmunología , Adulto , Anciano , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Femenino , Enfermedad de Graves/inmunología , Enfermedad de Hashimoto/inmunología , Humanos , Interleucina-10/sangre , Masculino , Persona de Mediana Edad , Estadísticas no Paramétricas , Pruebas de Función de la Tiroides , Glándula Tiroides/inmunología , Tirotropina/sangre , Tiroxina/sangre , Factor de Necrosis Tumoral alfa/sangreRESUMEN
Although studies have shown that 17beta-estradiol (estradiol) normalized Kupffer cell function following trauma-hemorrhage, the mechanism by which E2 maintains immune function remains unclear. Activation of Toll-like receptor 4 (TLR4) initiates an inflammatory cascade, involving activation of p38 mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K), and nuclear factor-kappaB (NF-kappaB). This leads to the release of proinflammatory cytokines. Thus, we hypothesized that the salutary effects of estradiol on Kupffer cell function following trauma-hemorrhage are mediated via negative regulation of TLR4-dependent p38 MAPK and NF-kappaB. TLR4 mutant (C3H/HeJ) and wild type (C3H/HeOuJ) mice were subjected to trauma-hemorrhage (mean BP 35+/-5 mmHg approximately 90 min, then resuscitation) or sham operation. Administration of estradiol following trauma-hemorrhage in wild type mice decreased Kupffer cell TLR4 expression as well as prevented the phosphorylation of p38 MAPK and NF-kappaB. This was accompanied by normalization of Kupffer cell production capacities of IL-6, TNF-alpha, macrophage inflammatory protein (MIP)-1alpha, and MIP-2 and the decrease in plasma cytokine levels. In contrast, TLR4 mutant mice did not exhibit the increase in Kupffer cell p38 MAPK and NF-kappaB activation, cytokine production, or the increase in circulating cytokine levels following trauma-hemorrhage. No difference was observed in activation of PI3K among groups. These results suggest that the protective effect of estradiol on Kupffer cell function is mediated via downregulation of TLR4-dependent p38 MAPK and NF-kappaB signaling following trauma-hemorrhage, which prevents the systemic release of cytokines.
Asunto(s)
Citocinas/biosíntesis , Regulación hacia Abajo/efectos de los fármacos , Estradiol/farmacología , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/enzimología , Receptor Toll-Like 4/genética , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CXCL2 , Citocinas/sangre , Hemorragia/inducido químicamente , Hemorragia/inmunología , Inflamación , Interleucina-6/biosíntesis , Interleucina-6/sangre , Proteínas Inflamatorias de Macrófagos/biosíntesis , Proteínas Inflamatorias de Macrófagos/sangre , Ratones , Monocinas/biosíntesis , Monocinas/sangre , FN-kappa B/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Fosforilación/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptor Toll-Like 4/metabolismo , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/sangre , Heridas y Lesiones/inducido químicamente , Heridas y Lesiones/inmunologíaRESUMEN
This study examines the effects of DEP components on circulatory CC and CXC chemokines, potent activators and chemoattractants for macrophage and leukocyte subpopulations, in a murine model of lung inflammation. ICR mice were divided into six experimental groups which received intratracheal inoculation of vehicle, LPS alone (2.5 mg/kg), organic chemicals in DEP (DEP-OC: 4 mg/kg) extracted with dichloromethane, residual carbonaceous nuclei after the extraction (washed DEP: 4 mg/kg), DEP-OC + LPS, or washed DEP + LPS. Intratracheal instillation of each DEP component alone did not significantly change the circulatory level of macrophage inflammatory protein (MIP)-1alpha, MIP-2, and macrophage chemoattractant protein-1 (MCP-1) 24 h after the exposure as compared with vehicle instilled alone. In the LPS group, MCP-1, but not MIP-1alpha or MIP-2, was significantly greater than in the vehicle group. The combined administration of LPS and washed DEP caused a further three to five-fold increase in MIP-1alpha, MIP-2, and MCP-1 proteins in the serum as compared with LPS administered alone. No significant difference between the LPS + DEP-OC group and the LPS group was observed. These results indicate that pulmonary exposure to washed DEP enhances circulatory level of chemokines during lung inflammation. The enhancement may be important in the aggravations of systemic inflammatory responses and ischemic cardiovascular conditions associated with air pollution.
Asunto(s)
Quimiocinas/sangre , Neumonía/inducido químicamente , Neumonía/metabolismo , Emisiones de Vehículos/toxicidad , Animales , Quimiocina CCL2/sangre , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CXCL2 , Quimiocinas CXC/sangre , Exposición por Inhalación , Lipopolisacáridos/farmacología , Proteínas Inflamatorias de Macrófagos/sangre , Masculino , Ratones , Ratones Endogámicos ICR , Monocinas/sangreRESUMEN
A water-soluble polysaccharide (EPA-1) from Pleurotus eryngii was obtained using DEAE-52 and Sephadex G-50 columns. The properties, structure and immunomodulatory activity of EPA-1 were studied. The results demonstrated that EPA-1 was a homogeneous polysaccharide with the molecular weight of 9.97×104Da. EPA-1 consisted of Man, Glc and Gal in a molar ratio of 2.2:1.0:3.2. The characterized fragment structures of EPA-1 were found to be consisting of seven sugar residues and two branches by GC-MS, FTIR and NMR analyses. Among them, the (1â6)-linkedGal residue was the main linkage mode of EpA-1 and composed of its backbone. Activity tests indicated that EPA-1 significantly induced macrophage to release the immune activity factor of NO, TNF-α, IL-1 and IL-6 through activation of signal protein of p38, ERK, JNK in MAPKs and translocation of nuclear NF-κΒ, indicating EPA-1 to possess good immunoregulatory activity.
Asunto(s)
Polisacáridos Fúngicos , Factores Inmunológicos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Macrófagos/metabolismo , Pleurotus/química , Animales , Conformación de Carbohidratos , Línea Celular Tumoral , Polisacáridos Fúngicos/química , Polisacáridos Fúngicos/aislamiento & purificación , Polisacáridos Fúngicos/farmacología , Factores Inmunológicos/química , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/farmacología , Sistema de Señalización de MAP Quinasas/inmunología , Macrófagos/inmunología , Masculino , Ratones , Monocinas/sangre , Monocinas/inmunología , Óxido Nítrico/inmunología , Óxido Nítrico/metabolismoRESUMEN
We have conducted a phase I study with autologous monocytes activated ex vivo and administered intraperitoneally in nine patients with peritoneal carcinomatosis. Blood monocytes were collected by leukapheresis and then purified by counterflow elutriation (up to 10(9) cells, with a purity of greater than 90%). Ex vivo activation was obtained by incubating these cells with 1 micrograms liposomal MTP-PE/10(6) monocytes for 18 hours in hydrophobic culture bags at 37 degrees C in 5% carbon dioxide humidified air. The activated monocytes were then infused in the peritoneal cavity once a week for 5 consecutive weeks through an implanted peritoneal infusion system, Port-A-Cath (Pharmacia Deltec, St Paul, MN), on an intrapatient dose-escalating schedule (10(7) to 10(9) monocytes). No severe adverse reactions occurred. Toxicity was mild, the chief acute reactions being fever (27%), chills (13%), and abdominal pain (25%). None of the side effects led to dose reduction. No consistent change in hemostatic function, liver function, or renal function was observed. Significant increases in granulocyte counts, neopterine, and acute phase reactants (fibrinogen, C-reactive protein) occurred in the peripheral blood. In vitro monocyte activation was demonstrated by the relapse of procoagulant activity and monokines (interleukin-1 [IL-1], IL-6, and tumor necrosis factor-alpha [TNF alpha]) in the supernatants of cultured monocytes. Evidence for in vivo monocyte activation was provided by the increase of these monokines in the peritoneal fluids. Kinetic studies with indium-111 (111In)-labeled activated autologous monocytes in five patients suggest that these infused monocytes may remain in the peritoneal cavity for up to 7 days. This locoregional immunotherapeutic approach seems to be encouraging in view of adjuvant therapeutic modality in ovarian cancer patients with minimal residual intraabdominal disease following second-look laparotomy.
Asunto(s)
Carcinoma/terapia , Monocitos Activados Asesinos , Neoplasias Peritoneales/terapia , Anciano , Análisis de Varianza , Recuento de Células Sanguíneas , Carcinoma/sangre , Carcinoma/etiología , Carcinoma/patología , Evaluación de Medicamentos , Femenino , Humanos , Radioisótopos de Indio , Infusiones Parenterales/instrumentación , Liposomas , Masculino , Persona de Mediana Edad , Monocinas/sangre , Neoplasias Peritoneales/sangre , Neoplasias Peritoneales/etiología , Neoplasias Peritoneales/patología , Análisis de Regresión , Factores de TiempoRESUMEN
Ischemia-reperfusion (I/R) injury is associated with activation of coagulation and inflammation. Interestingly, various anticoagulants have been shown to reduce both coagulation and inflammation in animal models of kidney I/R injury. Fondaparinux is a synthetic pentasaccharide that selectively inhibits factor Xa (FXa) in the coagulation cascade. The aim of this study was to investigate the effect of fondaparinux in a lethal murine model of kidney I/R injury. A murine model of kidney I/R was established. In this model, we measured activation of the coagulation cascade and induction of inflammation. Administration of fondaparinux to I/R-injured mice reduced fibrin deposition in the kidney, reduced serum creatinine levels and increased survival from 0 to 44% compared with saline-treated control mice. Fondaparinux also reduced interleukin-6 and macrophage inflammatory protein-2 expression and decreased neutrophil accumulation in the injured kidneys. Finally, we showed that fondaparinux reduced thioglycollate-induced recruitment of neutrophils into the peritoneum and inhibited the binding of U937 cells to P-selectin in vitro. Our data suggest that fondaparinux reduces kidney I/R injury primarily by inhibiting the recruitment of neutrophils.
Asunto(s)
Riñón/patología , Neutrófilos/efectos de los fármacos , Polisacáridos/farmacología , Daño por Reperfusión/tratamiento farmacológico , Animales , Coagulación Sanguínea/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Quimiocina CXCL2 , Creatina/sangre , Evaluación Preclínica de Medicamentos , Fibrina/metabolismo , Fondaparinux , Inflamación/tratamiento farmacológico , Interleucina-6/sangre , Riñón/irrigación sanguínea , Ratones , Modelos Animales , Monocinas/sangre , Polisacáridos/administración & dosificación , Tasa de SupervivenciaRESUMEN
Immunohistochemical double-label technique was used to detect trypanosomal antigen in macrophages. Immunoglobulin (Ig)M as well as IgG2a monoclonal antibodies (mAb) specific for the variant surface glycoprotein (VSG) mediated phagocytosis of Trypanosoma congolense variant antigenic type (VAT) TC13 by macrophages [bone marrow-derived macrophage cell line from BALB/c (BALB.BM)] in vitro. Administration of these IgM or IgG2a antibodies to BALB/c mice 30 min after injection of 3 x 10(8) T. congolense mediated phagocytosis of trypanosomes by Kupffer cells of the liver within 1 h. Plasma levels of the monokines interleukin (IL)-1beta, IL-10, and IL-12p40 were significantly increased 6-48 h after phagocytosis. In BALB/c mice infected with 10(3) T. congolense, a small degree of phagocytosis of trypanosomes by Kupffer cells, mediated by actively synthesized antibodies, was detected as early as 5 days after infection. Phagocytosis of trypanosomes was dramatically enhanced on day 6. Concomitantly, the Kupffer cells trippled in size. In BALB/c mice infected for 6 days, treatment with IgM or IgG2a mAb specific for T. congolense VSG led to clearance of VAT TC13 parasitemia but did not prevent death at the second parasitemia of a different VAT. We conclude that IgM as well as IgG antibody mediate phagocytosis of trypanosomes by Kupffer cells.
Asunto(s)
Macrófagos del Hígado/inmunología , Fagocitosis/inmunología , Trypanosoma congolense/inmunología , Tripanosomiasis Africana/inmunología , Animales , Inmunoglobulina G/inmunología , Inmunoglobulina M/inmunología , Macrófagos del Hígado/fisiología , Ratones , Monocinas/sangre , Fagocitosis/fisiología , Ratas , Factores de TiempoRESUMEN
The major proteoglycan in macrophages and platelets is the chondroitin sulphate proteoglycan serglycin. To study the biological role of serglycin, its binding to secreted and cell-associated proteins from macrophages and blood platelets was examined. Affinity chromatography with serglycin-Sepharose and chondroitin sulphate-Sepharose was used to isolate proteoglycin-binding proteins from macrophages and platelets. Antibodies against human macrophage inflammatory protein-1 alpha (MIP-1 alpha) precipitated a 14-kDa 35S-methionine-labeled protein among the chondroitin sulfate binding proteins secreted from the macrophage-like U937 cells after stimulation. Two proteins from murine macrophage J774 cells with molecular masses of approximately 10 and 14 kDa were precipitated by an antiserum against the murine MIP-1 alpha. Protein sequencing of fragments obtained by trypsin digestion of a 14-kDa chondroitin sulfate-binding protein from cell extracts of stimulated U937 cells revealed 100% homology with lysozyme, a bacteriolytic enzyme. Fragment of one other protein with approximate molecular mass of 8 kDa showed high homology with bone morphogenetic protein. Inhibition studies showed that chondroitin 6-sulfate inhibited the bacteriolytic activity of lysozyme in a competitive manner more efficiently than heparin and chondroitin 4-sulphate. Amino-terminal sequencing of two proteins from platelet extracts that bound to serglycin-Sepharose revealed that they corresponded to multimeric forms of human platelet factor 4 (PE4). Chondroitin sulfate-Sepharose was shown to be equally efficient in retaining PF4 from platelet extracts as serglycin-Sepharose indicating that the glycosaminoglycan chains mediate the binding to PF4 in the intact proteoglycan molecule. Competition experiments showed that serglycin was as efficient as heparin sulfate in blocking the binding of [3H] chondrotin sulfate to PF4, whereas heparin was one order of magnitude more efficient. Affinity measurements using fluoresceinamine-labeled glycosaminoglycans showed that the affinity of heparin for PF4 is on the order of 30 nM, whereas chondroitin sulfate has an affinity of 260 nM. Both PF4, MIP-1 alpha, and lysozyme play important role in different types of inflammatory reactions. The interaction with serglycin may indicate that this proteoglycan is involved in the regulation of the inflammatory response.
Asunto(s)
Plaquetas/metabolismo , Proteínas Sanguíneas/metabolismo , Macrófagos/metabolismo , Monocinas/sangre , Factor Plaquetario 4/metabolismo , Proteoglicanos/sangre , Animales , Quimiocina CCL4 , Sulfatos de Condroitina/sangre , Cromatografía de Afinidad , Heparina/sangre , Humanos , Activación de Macrófagos/fisiología , Proteínas Inflamatorias de Macrófagos , Ratones , Muramidasa/sangre , Pruebas de Precipitina , Unión Proteica , Tritio , Proteínas de Transporte VesicularRESUMEN
We hypothesised that circulating monocytes of women with vaginal colonisation with Ureaplasma spp., genital microorganisms known to cause inflammation-driven preterm birth, would elicit a tolerised cytokine response to subsequent in vitro Ureaplasma parvum serovar 3 (UpSV3) stimulation. Using multi-parameter flow cytometry, we found no differences with regard to maternal colonisation status in the frequency of TNF-α-, IL-6-, IL-8- and IL-1ß-expressing monocytes in response to subsequent UpSV3 stimulation (P > 0.10 for all cytokines). We conclude that vaginal Ureaplasma spp. colonisation does not specifically tolerise monocytes of pregnant women towards decreased responses to subsequent stimulation.
Asunto(s)
Citometría de Flujo , Monocitos/inmunología , Monocinas/inmunología , Ureaplasma/inmunología , Adulto , Femenino , Humanos , Monocitos/metabolismo , Monocinas/sangre , Embarazo , Nacimiento Prematuro/sangre , Nacimiento Prematuro/inmunología , Nacimiento Prematuro/microbiologíaRESUMEN
Hyperlipidaemia may accelerate the development of atherosclerosis by enhancing the expression of chemokines by cells within the arterial wall. Chemokines of the CC subfamily are clearly implicated in atherogenesis; however, recent reports suggest that CXC chemokines may play a hitherto unrecognised role in monocyte recruitment into atheromatous lesions expressing these molecules. Here, we examine whether circulating levels of CXC chemokines may reflect the pathogenic changes occurring during early atherogenesis. ApoE*3 Leiden mice developed marked hypercholesterolaemia, and early Type I 'fatty streak' lesions, following consumption of an atherogenic diet high in saturated fat and cholesterol, and containing sodium cholate, for up to 4 weeks. By contrast, their non-transgenic littermates (C57BL/6J) exhibited a much less pronounced hypercholesterolaemia and did not develop fatty streak lesions, when fed the same diet. Under these conditions, serum concentrations of CXC chemokines, KC and Macrophage Inflammatory Protein-2 (MIP-2) were significantly (PAsunto(s)
Arteriosclerosis/patología
, Quimiocinas CXC/sangre
, Dieta Aterogénica
, Hipercolesterolemia/sangre
, Monocinas/sangre
, ARN Mensajero/análisis
, Animales
, Aorta/patología
, Apolipoproteínas E
, Arteriosclerosis/fisiopatología
, Secuencia de Bases
, Quimiocina CXCL2
, Quimiocinas CXC/metabolismo
, Modelos Animales de Enfermedad
, Femenino
, Hipercolesterolemia/fisiopatología
, Inmunohistoquímica
, Masculino
, Ratones
, Ratones Transgénicos
, Datos de Secuencia Molecular
, Monocinas/metabolismo
, Reacción en Cadena de la Polimerasa
, Probabilidad
, Valores de Referencia
, Factores de Riesgo
, Sensibilidad y Especificidad
RESUMEN
Flagellin is a recently identified bacterial product that elicits immune response via toll-like receptor 5. Here, we demonstrate that flagellin is an extraordinarily potent proinflammatory stimulus in the lung during sepsis. In vitro, flagellin triggers the production of interleukin (IL)-8 by human lung epithelial (A549) cells, with 50% of the maximal response obtained at a concentration of 2 x 10(-14) M. Flagellin also induces the expression of ICAM-1 in vitro. Intravenous administration of flagellin to mice elicited a severe acute lung inflammation that was significantly more pronounced than following lipopolysaccharide (LPS) administration. Flagellin induced a local release of proinflammatory cytokines, the accumulation of inflammatory cells, and the development of pulmonary hyperpermeability. These effects were associated with the nuclear translocation of the transcription NF-kappaB in the lung. Flagellin remained active in inducing pulmonary inflammation at doses as low as 10 ng/mouse. In the plasma of patients with sepsis, flagellin levels amounted to 7.1 +/- 0.1 ng/mL. Plasma flagellin levels showed a significant positive correlation with the lung injury score, with the alveolar-arterial oxygen difference as well as with the duration of the sepsis. Flagellin emerges as a potent trigger of acute respiratory complications in gram-negative bacterial sepsis.
Asunto(s)
Flagelina/metabolismo , Bacterias Gramnegativas/metabolismo , Inflamación/metabolismo , Pulmón/inmunología , Sepsis/inmunología , Transporte Activo de Núcleo Celular , Animales , Células Cultivadas , Quimiocina CCL4 , Quimiocina CXCL2 , Quimiocinas/metabolismo , Relación Dosis-Respuesta a Droga , Flagelina/sangre , Humanos , Inflamación/microbiología , Interleucina-1/sangre , Interleucina-8/metabolismo , Lipopolisacáridos/sangre , Pulmón/metabolismo , Pulmón/microbiología , Proteínas Inflamatorias de Macrófagos/sangre , Masculino , Ratones , Ratones Endogámicos BALB C , Monocinas/sangre , FN-kappa B/metabolismo , Neutrófilos/metabolismo , Óxido Nítrico/metabolismo , Salmonella/metabolismo , Sepsis/metabolismo , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
The effect of (R)-1-(5-hydroxyhexyl)-3,7-dimethylxanthine (CT-1501R; the nonproprietary name for CT-1501R approved by the United States Name Council is lisofylline), an inhibitor of second messenger signaling through phosphatidic acid, on release of endogenous mediators important in the systemic inflammatory response syndrome (SIRS) was studied using the human whole blood ex vivo assay system. Human blood was stimulated with various endotoxin preparations, zymosan, or protein A, and the levels of secreted monokines were measured by enzyme-linked immunosorbent assay. CT-1501R inhibited tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta), and IL-6 release in a dose-dependent manner and was active with all stimuli tested including Salmonella and Escherichia coli-derived endotoxin, endotoxin from both rough and smooth E. coli strains, as well as zymosan and protein A. CT-1501R inhibited monokine release by approximately 50% at 200 microM and 30% at 50 microM and was independent of the relative potency of stimulus. CT-1501R also inhibited IL-1 alpha or IL-1 beta induction of either TNF-alpha or IL-1 beta and inhibited the synergistic effects of stimulation with both human IL-1 beta and murine TNF-alpha on release of human TNF-alpha. Inhibition of monokine release following stimulation with monokine(s) was, in general, greater than that achieved with lipopolysaccharide (LPS) stimulation. Northern blot analysis showed decreased mRNA accumulation of TNF-alpha and IL-1 beta in CT-1501R-treated samples following LPS stimulation suggesting that CT-1501R acts at least in part, at the pretranslational level. In contrast, CT-1501R does not inhibit LPS-stimulated IL-8 or IL-1 receptor antagonist (IL-1ra) release in human whole blood or IL-1 alpha-induced release of PGE2 in human foreskin fibroblast cells. These data suggest that CT-1501R may be of use for clinical intervention in SIRS.
Asunto(s)
Inflamación/prevención & control , Monocinas/antagonistas & inhibidores , Monocinas/sangre , Pentoxifilina/análogos & derivados , Células 3T3 , Animales , Células Cultivadas , Dinoprostona/biosíntesis , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Técnicas In Vitro , Inflamación/sangre , Inflamación/etiología , Proteína Antagonista del Receptor de Interleucina 1 , Interleucina-1/genética , Interleucina-1/metabolismo , Interleucina-1/farmacología , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Ratones , Pentoxifilina/farmacología , Ácidos Fosfatidicos/biosíntesis , ARN Mensajero/sangre , ARN Mensajero/genética , Sialoglicoproteínas/metabolismo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
BACKGROUND: Previous clinical reports have documented multisystem organ injury after hepatic cryoablation. We hypothesized that hepatic cryosurgery, but not partial hepatectomy, induces a systemic inflammatory response characterized by distant organ injury and overproduction of nuclear factor kappa B (NF-kappa B)-dependent, proinflammatory cytokines. METHODS: In this study, rats underwent either cryoablation of 35% of liver parenchyma or a similar resection of left hepatic tissue. Serum tumor necrosis factor-alpha and macrophage inflammatory protein-2 levels and NF-kappa B activation were assessed by electrophoretic mobility shift assay at 30 minutes 1, 2, 6, and 24 hours after either procedure. RESULTS: Cryoablation of 35% of liver (n = 22 rats) resulted in lung injury and a 45% mortality rate within 24 hours of surgery, whereas 7% treated with 35% hepatectomy (n = 15 rats) died during the 24 hours after surgery (P < .05, cryoablation vs hepatectomy). Serum tumor necrosis factor-alpha and macrophage inflammatory protein-2 levels were markedly increased in rats (n = 10 rats) 1 hour after hepatic cryoablation compared with rats that underwent partial hepatectomy (P < .005). We evaluated NF-kappa B activation by electrophoretic mobility shift assay in nuclear extracts of liver and lung after cryosurgery and found that NF-kappa B activation was strikingly increased in the liver but not the lung at 30 minutes and in both organs 1 hour after cryosurgery, and returned to baseline in both organs by 2 hours. In rats undergoing 35% hepatectomy, no increase in NF-kappa B activation was detected in nuclear extracts of either liver or lung at any time point. CONCLUSIONS: These data show that hepatic cryosurgery results in systemic inflammation with activation of NF-kappa B and increased production of NF-kappa B-dependent cytokines. Our data suggest that lung injury and death in this animal model is mediated by an exaggerated inflammatory response to cryosurgery.
Asunto(s)
Criocirugía/efectos adversos , Citocinas/biosíntesis , Hígado/cirugía , Lesión Pulmonar , FN-kappa B/metabolismo , Enfermedad Aguda , Animales , Quimiocina CXCL2 , Modelos Animales de Enfermedad , Hepatectomía/efectos adversos , Humanos , Mediadores de Inflamación/metabolismo , Hígado/metabolismo , Pulmón/metabolismo , Pulmón/patología , Monocinas/sangre , Ratas , Ratas Sprague-Dawley , Factores de Tiempo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE: To determine whether closed bone fracture in conjunction with hemorrhagic shock compromises immune functions more severely than hemorrhagic shock alone. DESIGN: In a randomized, controlled trial, closed bone fracture of the right lower leg and/or hemorrhagic shock (mean +/- SEM arterial blood pressure, 35 +/- 5 mm Hg for 90 minutes) were induced in male C3H/HeN mice (weight, 25 g). Animals subjected to hemorrhage were resuscitated with the shed blood and lactated Ringer solution. At 72 hours after the experiment, all animals were killed to obtain whole blood, splenocytes, and splenic and peritoneal macrophages. Macrophage interleukin-1 and splenocyte interleukin-2 and interleukin-3 release were determined by bioassay, and splenocyte proliferation was measured by tritiated thymidine incorporation. RESULTS: Closed bone fracture alone did not affect immune functions 72 hours after the trauma. Hemorrhagic shock, however, induced a significant depression of splenocyte and macrophage functions. Bone fracture followed by hemorrhagic shock further depressed splenocyte proliferation and splenocyte interleukin-2 and interleukin-3 release as well as interleukin-1 release. CONCLUSION: Since bone injury coupled with hemorrhagic shock produces more severe depression of immune functions than hemorrhage alone, bone injury appears to play a contributory role in further depressing immune functions in trauma patients who experience major blood loss.
Asunto(s)
Fracturas Cerradas/inmunología , Choque Hemorrágico/inmunología , Animales , Fracturas Cerradas/sangre , Fracturas Cerradas/complicaciones , Linfocinas/sangre , Masculino , Ratones , Ratones Endogámicos C3H , Monocinas/sangre , Choque Hemorrágico/sangre , Choque Hemorrágico/complicacionesRESUMEN
Many findings demonstrated that Glycosaminoglycans (GAGs) and Proteoglycans (PGs) possess antioxidant activity. Collagen-induced arthritis (CIA) is an experimental animal model similar to human rheumatoid arthritis (RA) in which free radicals are involved. Sodium salicylate can be used as a chemical trap for hydroxyl radicals (OH*), the most damaging reactive oxygen species (ROS), yielding 2,5-dihydroxybenzoic acid), (2,5-DHBA) and 2,3-dihydroxybenzoic acid (2,3-DHBA). The measurement of these two acids in the plasma allows to indirectly assess the production of OH* radicals. The aim of the study was to investigate the effect of hyaluronic acid (HYA) (30 mg/kg i.p.) or chondroitin-4-sulphate (C4S) (30 mg/kg i.p.), on free radical production in Lewis rats subjected to CIA. After the immunization with bovine collagen type II in complete Freund's adjuvant, rats developed an erosive hind paw arthritis, that produced high plasma OH* levels assayed as 2,3-DHBA and 2,5-DHBA, primed lipid peroxidation, evaluated by analyzing conjugated dienes (CD) in the articular cartilage; decreased the concentration of endogenous vitamin E (VE) and catalase (CA) in the joint cartilage; enhanced macrophage inflammatory protein-2 (MIP-2) serum levels and increased elastase (ELA) evaluated as an index of activated leukocyte polymophonuclear (PMNs) accumulation in the articular joints. The administration of HYA and C4S starting at the onset of arthritis (day 11) for 20 days, limited inflammation and the clinical signs in the knee and paw, reduced OH* production, decreased CD levels, partially restored the endogenous antioxidants VE and CA, reduced MIP-2 serum levels and limited PMNs infiltration. The results indicate that the GAGs HYA and C4S significantly reduce free radical production in CIA and could be used as a tool to investigate the role of antioxidants in RA.