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Immune response dysregulation plays a key role in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pathogenesis. In this study, we evaluated immune and endothelial blood cell profiles of patients with coronavirus disease 2019 (COVID-19) to determine critical differences between those with mild, moderate, or severe COVID-19 using spectral flow cytometry. We examined a suite of immune phenotypes, including monocytes, T cells, NK cells, B cells, endothelial cells, and neutrophils, alongside surface and intracellular markers of activation. Our results showed progressive lymphopenia and depletion of T cell subsets (CD3+, CD4+, and CD8+) in patients with severe disease and a significant increase in the CD56+CD14+Ki67+IFN-γ+ monocyte population in patients with moderate and severe COVID-19 that has not been previously described. Enhanced circulating endothelial cells (CD45-CD31+CD34+CD146+), circulating endothelial progenitors (CD45-CD31+CD34+/-CD146-), and neutrophils (CD11b+CD66b+) were coevaluated for COVID-19 severity. Spearman correlation analysis demonstrated the synergism among age, obesity, and hypertension with upregulated CD56+ monocytes, endothelial cells, and decreased T cells that lead to severe outcomes of SARS-CoV-2 infection. Circulating monocytes and endothelial cells may represent important cellular markers for monitoring postacute sequelae and impacts of SARS-CoV-2 infection during convalescence and for their role in immune host defense in high-risk adults after vaccination.
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COVID-19/inmunología , Células Endoteliales/inmunología , Monocitos/inmunología , SARS-CoV-2 , Adolescente , Adulto , Factores de Edad , Anciano , Anticuerpos Antivirales/biosíntesis , Anticuerpos Antivirales/inmunología , Biomarcadores , Antígeno CD56/análisis , COVID-19/sangre , COVID-19/epidemiología , Niño , Comorbilidad , Células Endoteliales/química , Femenino , Citometría de Flujo , Humanos , Hipertensión/epidemiología , Hipertensión/inmunología , Inmunofenotipificación , Activación de Linfocitos , Subgrupos Linfocitarios/inmunología , Linfopenia/etiología , Linfopenia/inmunología , Masculino , Persona de Mediana Edad , Monocitos/química , Neutrófilos/inmunología , Obesidad/epidemiología , Obesidad/inmunología , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/análisis , SARS-CoV-2/inmunología , Índice de Severidad de la Enfermedad , Glicoproteína de la Espiga del Coronavirus/inmunología , Adulto JovenRESUMEN
Background Today, there is a need for new and independent additional advanced markers that can predict the prognosis of meningioma patients, postoperatively. The present study aimed to find out postoperative short-term prognostic markers in patients with meningioma using their demographic data and routine blood biochemistry findings evaluated preoperatively. Methods The Glasgow Coma Scale (GCS), and Glasgow Outcome Scale (GOS) scores of the patients were recorded. Additionally, preoperatively obtained serum glucose, Creactive protein (CRP), sodium, potassium, creatinine, blood urea nitrogen, aspartate aminotransferase (AST), alanine aminotransferase, and hemoglobin level values, platelet, leukocyte, neutrophil, lymphocyte, eosinophil, basophil, andmonocyte count results, erythrocyte sedimentation rate (ESR), neutrophil-lymphocyte ratio, plateletlymphocyte ratio (PLR) and lymphocyte-monocyte ratio (LMR) values were evaluated. Results In the present study, 23 operated patients with meningioma World Health Organization (WHO) grade 1 (17 females, 6 males) were included. Correlation test results revealed that the GCS score, platelet count, and serum potassium level values could directly predict the short-term prognosis of these patients. Additionally, these test results suggested that the lymphocyte, monocyte, and eosinophil count values, PLR, LMR, ESR, serum glucose, CRP, and AST level values could be indirect markers in predicting the short-term prognosis. However, likelihood ratio test results revealed that only monocyte count value, LMR value, and serum CRP level value could be the markers for prediction of the short-term prognosis. Conclusion At the end of the present study, it was concluded that the monocyte count value, LMR value, and serum CRP level value could be the best markers in predicting the short-term prognosis of the operated meningioma patients.
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Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Anciano , Análisis Químico de la Sangre/métodos , Biomarcadores , Meningioma/terapia , Recuento de Plaquetas , Potasio/sangre , Pronóstico , Proteína C-Reactiva/química , Monocitos/química , Estudios Retrospectivos , Interpretación Estadística de Datos , Estadísticas no Paramétricas , Correlación de DatosRESUMEN
Introducción: La lesión central (LCCG) y periférica (LPCG) de células gigantes de los maxilares, son lesiones reactivas con comportamiento clínico diferente. Objetivo: Comparar la inmunoexpresión de CD68 en células gigantes (CGm) mononucleares (CMn) en lesiones central y periférica de los maxilares. Material y métodos: Se evaluaron 35 casos de LCCG y 24 de LPCG en bloques de parafi na que podían ser procesadas para la expresión del anticuerpo CD68. La inmunoexpresión se valoró en el citoplasma de ambas poblaciones celulares, obteniendo proporciones; la inmunoexpresión se categorizó en intensa, moderada, leve. Las proporciones se compararon con χ2, siendo signifi cativo p ≤ 0.05. Resultados: Para las CGm de LCCG, CD68 se expresó en una proporción de 96 versus 84.2% LPCG (p < 0.005). La proporción de la tinción de la expresión intensa y moderada fue más frecuente en las LCCG (p = 0.032). Las proporciones entre las CMn 59.3% LCCG versus 18.6% en la LPCG (p < 0.001). Hubo diferencia en intensidad de CD68, en las CMn de LCCG fue mayor (p = 0.002). Conclusiones: La alta expresión de CD68 en las CGM y CMn en la lesión central y periférica confi rma su fenotipo de macrófago. Las diferencias entre las proporciones y la tinción a CD68 refl eja mayor actividad fagocítica posiblemente relacionada con el comportamiento clínico (AU)
Introduction: Central (CGCL) and Peripheral (PGCL) giant cell lesions of jaws are reactive lesions displaying diff erent behavior patterns. Objective: To compare CD68 immunoexpression between CGCL and PCGL in giant multinucleated and mononuclear cells. Material and methods: 35 CGCL and 24 PGCL were retrieved from paraffi n-embedded biopsy, as well as the feasibility to analyze CD68 immunoexpression. The immunoexpression was analyzed in cytoplasm both cell populations cellular, for and staining intensity was categorized as intense, moderate or faint. Proportions were compared by χ2, making a p ≤ 0.05 value signifi cate. Results: In 96% of CGCL's in GMCs displayed CD68, as compared to 84.2% in PGCL, (p < 0.005). The proportion of stained cells, intense to moderate staining was more frequent in CGCL (p = 0.032). The proportion CD68 was expressed in 59.3% or CGCL mononuclear cells, as compared to 18.6% in PGCL, (p < 0.001). There was diff erence in staining CD68 intensity between mononuclear cells in CGCL, (p = 0.002). Conclusions: The high CD68 expression frequency in GMCs and mononuclear cells in central and peripheral GCL confi rm a macrophage phenotype; a more intense staining in CGML and GMCs suggests a more active phagocytic activity, and possibility underline the diff erent clinical behavior (AU)
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Niño , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Inmunohistoquímica , Granuloma de Células Gigantes/genética , Enfermedades Maxilomandibulares/inmunología , Antígenos CD , Monocitos/química , Interpretación Estadística de Datos , Distribución por Edad y Sexo , Macrófagos/química , MéxicoRESUMEN
OBJECTIVE: To observe the effect of short-term insulin intensive treatment on the monocyte chemoattractant protein-1 (MCP-1) as well as on the nuclear factor-kappa B (NF-κB) expression of peripheral blood monocyte. This is also in addition to observing the serum MCP-1 level in newlydiagnosed type 2 diabetic patients and probing its anti-inflammation effects. SUBJECTS AND METHODS: Twenty newly-diagnosed type 2 diabetic patients were treated with an insulin intensive treatment for 2 weeks. MCP-1 and NF-κB expression on the monocyte surface were measured with flow cytometry, the serum MCP-1 level was measured by enzyme linked immunosorbent assay (ELISA) during pretreatment and post-treatment. RESULTS: After 2 weeks of the treatment, MCP-1 and NF-κB protein expression of peripheral blood monocyte and serum MCP-1 levels decreased significantly compared with those of pre-treatment, which were (0.50 ± 0.18)% vs (0.89 ± 0.26)% (12.22 ± 2.80)% vs (15.53 ± 2.49)% and (44.53 ± 3.97) pg/mL vs (49.53 ± 3.47) pg/mL, respectively (P < 0.01). The MCP-1 expression on monocyte surface had a significant positive relationship with serum MCP-1 levels (r = 0.47, P < 0.01). CONCLUSIONS: Short-term insulin intensive therapy plays a role in alleviating the increased inflammation reaction in type 2 diabetics.
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Quimiocina CCL2/efectos de los fármacos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Inflamación/prevención & control , Insulina/administración & dosificación , Monocitos/química , FN-kappa B/efectos de los fármacos , Estudios de Casos y Controles , Quimiocina CCL2/sangre , Diabetes Mellitus Tipo 2/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Humanos , Masculino , Persona de Mediana Edad , FN-kappa B/sangreRESUMEN
ABSTRACT Objective To observe the effect of short-term insulin intensive treatment on the monocyte chemoattractant protein-1 (MCP-1) as well as on the nuclear factor-kappa B (NF-κB) expression of peripheral blood monocyte. This is also in addition to observing the serum MCP-1 level in newlydiagnosed type 2 diabetic patients and probing its anti-inflammation effects. Subjects and methods Twenty newly-diagnosed type 2 diabetic patients were treated with an insulin intensive treatment for 2 weeks. MCP-1 and NF-κB expression on the monocyte surface were measured with flow cytometry, the serum MCP-1 level was measured by enzyme linked immunosorbent assay (ELISA) during pretreatment and post-treatment. Results After 2 weeks of the treatment, MCP-1 and NF-κB protein expression of peripheral blood monocyte and serum MCP-1 levels decreased significantly compared with those of pre-treatment, which were (0.50 ± 0.18)% vs (0.89 ± 0.26)% (12.22 ± 2.80)% vs (15.53 ± 2.49)% and (44.53 ± 3.97) pg/mL vs (49.53 ± 3.47) pg/mL, respectively (P < 0.01). The MCP-1 expression on monocyte surface had a significant positive relationship with serum MCP-1 levels (r = 0.47, P < 0.01). Conclusions Short-term insulin intensive therapy plays a role in alleviating the increased inflammation reaction in type 2 diabetics.
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Humanos , Masculino , Femenino , Persona de Mediana Edad , Monocitos/química , FN-kappa B/efectos adversos , Quimiocina CCL2/efectos de los fármacos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Inflamación/prevención & control , Insulina/administración & dosificación , Ensayo de Inmunoadsorción Enzimática , Estudios de Casos y Controles , FN-kappa B/sangre , Quimiocina CCL2/sangre , Diabetes Mellitus Tipo 2/sangre , Citometría de FlujoAsunto(s)
Biomarcadores/análisis , Enfermedades Cardiovasculares/epidemiología , Transportador de Glucosa de Tipo 1/análisis , Infecciones por VIH/complicaciones , Infecciones por VIH/patología , Monocitos/química , Adulto , Fármacos Anti-VIH/uso terapéutico , Australia/epidemiología , Infecciones por VIH/tratamiento farmacológico , Humanos , Medición de Riesgo , Respuesta Virológica SostenidaRESUMEN
BACKGROUND & AIMS: Resident macrophages are derived from yolk sac precursors and seed the liver during embryogenesis. Native cells may be replaced by bone marrow precursors during extensive injuries, irradiation, and infections. We investigated the liver populations of myeloid immune cells and their location, as well as the dynamics of phagocyte repopulation after full depletion. The effects on liver function due to the substitution of original phagocytes by bone marrow-derived surrogates were also examined. METHODS: We collected and analyzed liver tissues from C57BL/6 (control), LysM-EGFP, B6 ACTb-EGFP, CCR2-/-, CD11c-EYFP, CD11c-EYFP-DTR, germ-free mice, CX3CR1gfp/gfp, CX3CR1gpf/wt, and CX3CR1-DTR-EYFP. Liver nonparenchymal cells were immunophenotyped using mass cytometry and gene expression analyses. Kupffer and dendritic cells were depleted from mice by administration of clodronate, and their location and phenotype were examined using intravital microscopy and time-of-flight mass cytometry. Mice were given acetaminophen gavage or intravenous injections of fluorescently labeled Escherichia coli, blood samples were collected and analyzed, and liver function was evaluated. We assessed cytokine profiles of liver tissues using a multiplexed array. RESULTS: Using mass cytometry and gene expression analyses, we identified 2 populations of hepatic macrophages and 2 populations of monocytes. We also identified 4 populations of dendritic cells and 1 population of basophils. After selective depletion of liver phagocytes, intravascular myeloid precursors began to differentiate into macrophages and dendritic cells; dendritic cells migrated out of sinusoids, after a delay, via the chemokine CX3CL1. The cell distribution returned to normal in 2 weeks, but the repopulated livers were unable to fully respond to drug-induced injury or clear bacteria for at least 1 month. This defect was associated with increased levels of inflammatory cytokines, and dexamethasone accelerated the repopulation of liver phagocytes. CONCLUSIONS: In studies of hepatic phagocyte depletion in mice, we found that myeloid precursors can differentiate into liver macrophages and dendritic cells, which each localize to distinct tissue compartments. During replenishment, macrophages acquire the ability to respond appropriately to hepatic injury and to remove bacteria from the blood stream.
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Antígenos CD/análisis , Células de la Médula Ósea/fisiología , Diferenciación Celular , Hígado/citología , Hígado/fisiopatología , Células Mieloides/fisiología , Acetaminofén , Animales , Células de la Médula Ósea/citología , Enfermedad Hepática Inducida por Sustancias y Drogas/inmunología , Quimiocina CX3CL1/metabolismo , Citocinas/genética , Citocinas/metabolismo , Células Dendríticas/química , Inmunofenotipificación/métodos , Microscopía Intravital , Lectinas/genética , Hígado/inmunología , Hígado/metabolismo , Macrófagos/química , Macrófagos/inmunología , Ratones , Ratones Endogámicos C57BL , Microscopía Confocal , Microvasos/metabolismo , Monocitos/química , Células Mieloides/química , Fenotipo , TranscriptomaRESUMEN
BACKGROUND & AIMS: Hypovitaminosis D and inflammation are highly prevalent among patients undergoing dialysis, and the association of both conditions with worse survival has been well recognized. Although a potential role for vitamin D in the immune system has been suggested, the effect of the treatment of hypovitaminosis D on the modulation of the inflammatory response remains unclear. The aim of this study was to investigate the effect of the restoration of the vitamin D status on the expression of vitamin D-regulatory proteins in monocytes and on circulating inflammatory markers in dialysis patients. METHODS: In this randomized double-blind placebo-controlled 12-week trial, 38 patients on dialysis with serum 25-hydroxyvitamin D [25(OH)D] <20 ng/mL were randomized either to the cholecalciferol group (n = 20; 50,000 IU of cholecalciferol twice weekly) or to the control group (n = 18; 50 drops of a placebo solution twice weekly). The expression of vitamin D receptor (VDR), CYP27B1, CYP24A1 and interleukin-6 (IL-6) in monocytes was determined by flow cytometry. Serum concentrations of 25(OH)D, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) were measured. The trial is registered at ClinicalTrials.gov #NCT01974245. RESULTS: After 12 weeks, the serum 25(OH)D increased from 14.3 ± 4.7 ng/mL to 43.1 ± 11.0 ng/mL (p < 0.05) in the cholecalciferol group and did not change in the control group (13.9 ± 4.2 ng/mL to 13.5 ± 4.3 ng/mL; p = 0.56). In monocytes, while CYP27B1 expression and VDR expression increased in the cholecalciferol group (p < 0.05), CYP27B1 expression did not change, and VDR expression decreased in the control group (p < 0.05). There were no changes in IL-6 and CYP24A1 expression in both groups. Serum concentration of IL-6 and CRP decreased from 8.1 ± 6.6 pg/mL to 4.6 ± 4.1 pg/mL (p < 0.05) and from 0.50 (0.10-1.27) mg/dL to 0.28 (0.09-0.62) mg/dL (p < 0.05), respectively only in the cholecalciferol group. Assessed overtime, the treatment group differences in 25(OH) D, PTH, CRP and IL-6, CYP27B1 and VDR remained significant. CONCLUSIONS: Restoration of vitamin D status of patients undergoing dialysis promoted upregulation of CYP27B1 and VDR expression in monocytes and a decrease in circulating inflammatory markers.
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Biomarcadores/sangre , Inflamación/sangre , Monocitos/química , Diálisis Renal , Deficiencia de Vitamina D/tratamiento farmacológico , Vitamina D/análogos & derivados , 25-Hidroxivitamina D3 1-alfa-Hidroxilasa/sangre , Adulto , Anciano , Brasil , Proteína C-Reactiva/análisis , Colecalciferol/administración & dosificación , Método Doble Ciego , Femenino , Humanos , Interleucina-6/sangre , Masculino , Persona de Mediana Edad , Placebos , Receptores de Calcitriol/sangre , Vitamina D/sangre , Deficiencia de Vitamina D/sangre , Vitamina D3 24-Hidroxilasa/sangreRESUMEN
UNLABELLED: High levels of circulating immunocomplexes (ICs) are found in patients with either infectious or sterile inflammation. We report that patients with either Plasmodium falciparum or Plasmodium vivax malaria have increased levels of circulating anti-DNA antibodies and ICs containing parasite DNA. Upon stimulation with malaria-induced ICs, monocytes express an NF-κB transcriptional signature. The main source of IC-induced proinflammatory cytokines (i.e., tumor necrosis factor alpha [TNF-α] and interleukin-1ß [IL-1ß])in peripheral blood mononuclear cells from acute malaria patients was found to be a CD14(+) CD16 (FcγRIIIA)(+) CD64 (FcγRI)(high) CD32 (FcγRIIB)(low) monocyte subset. Monocytes from convalescent patients were predominantly of the classical phenotype (CD14(+) CD16(-)) that produces high levels of IL-10 and lower levels of TNF-α and IL-1ß in response to ICs. Finally, we report a novel role for the proinflammatory activity of ICs by demonstrating their ability to induce inflammasome assembly and caspase-1 activation in human monocytes. These findings illuminate our understanding of the pathogenic role of ICs and monocyte subsets and may be relevant for future development of immunity-based interventions with broad applications to systemic inflammatory diseases. IMPORTANCE: Every year, there are approximately 200 million cases of Plasmodium falciparum and P. vivax malaria, resulting in nearly 1 million deaths, most of which are children. Decades of research on malaria pathogenesis have established that the clinical manifestations are often a consequence of the systemic inflammation elicited by the parasite. Recent studies indicate that parasite DNA is a main proinflammatory component during infection with different Plasmodium species. This finding resembles the mechanism of disease in systemic lupus erythematosus, where host DNA plays a central role in stimulating an inflammatory process and self-damaging reactions. In this study, we disclose the mechanism by which ICs containing Plasmodium DNA activate innate immune cells and consequently stimulate systemic inflammation during acute episodes of malaria. Our results further suggest that Toll-like receptors and inflammasomes have a central role in malaria pathogenesis and provide new insights toward developing novel therapeutic interventions for this devastating disease.
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Complejo Antígeno-Anticuerpo/metabolismo , Citocinas/metabolismo , ADN Protozoario/inmunología , Inflamasomas/metabolismo , Malaria Falciparum/patología , Malaria Vivax/patología , Monocitos/metabolismo , Complejo Antígeno-Anticuerpo/sangre , Antígenos CD/análisis , Humanos , Inmunofenotipificación , Malaria Falciparum/inmunología , Malaria Vivax/inmunología , Monocitos/química , Multimerización de ProteínaRESUMEN
Ulcer development in patients with cutaneous leishmaniasis (CL) caused by Leishmania braziliensis is associated with high levels of tumor necrosis factor (TNF). We found that early after infection, before ulcer development, the frequency of CD16(+) (both intermediate [CD14(+)CD16(+)] and nonclassical [CD14(dim)CD16(+)]) monocytes was increased in the peripheral blood of patients with L. braziliensis, compared with uninfected controls. These results suggest that CD16(+) monocytes might promote disease. Also, we found that intermediate monocytes expressed CCR2 and that increased levels of CCL2 protein were present in lesions from patients, suggesting that intermediate monocytes are more likely than nonclassical monocytes to migrate to the lesion site. Finally, we found that the intermediate monocytes produced TNF. Our results show that intermediate monocytes are increased in frequency soon after infection; express CCR2, which would promote their migration into the lesions; and, owing to their production of TNF, can enhance the inflammatory response.
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Leishmania braziliensis/inmunología , Leishmaniasis Cutánea/inmunología , Leishmaniasis Cutánea/patología , Monocitos/inmunología , Adolescente , Adulto , Quimiocina CCL2/metabolismo , Femenino , Proteínas Ligadas a GPI/análisis , Humanos , Masculino , Persona de Mediana Edad , Monocitos/química , Receptores CCR2/análisis , Receptores de IgG/análisis , Factor de Necrosis Tumoral alfa/metabolismo , Úlcera/inmunología , Úlcera/patología , Adulto JovenRESUMEN
Recognition of pathogens is performed by specific receptors in cells of the innate immune system, which may undergo modulation during the continuum of clinical manifestations of sepsis. Monocytes and neutrophils play a key role in host defense by sensing and destroying microorganisms. This study aimed to evaluate the expression of CD14 receptors on monocytes; CD66b and CXCR2 receptors on neutrophils; and TLR2, TLR4, TLR5, TLR9, and CD11b receptors on both cell types of septic patients. Seventy-seven septic patients (SP) and 40 healthy volunteers (HV) were included in the study, and blood samples were collected on day zero (D0) and after 7 days of therapy (D7). Evaluation of the cellular receptors was carried out by flow cytometry. Expression of CD14 on monocytes and of CD11b and CXCR2 on neutrophils from SP was lower than that from HV. Conversely, expression of TLR5 on monocytes and neutrophils was higher in SP compared with HV. Expression of TLR2 on the surface of neutrophils and that of TLR5 on monocytes and neutrophils of SP was lower at D7 than at D0. In addition, SP who survived showed reduced expression of TLR2 and TLR4 on the surface of neutrophils at D7 compared to D0. Expression of CXCR2 for surviving patients was higher at follow-up compared to baseline. We conclude that expression of recognition and cell signaling receptors is differentially regulated between SP and HV depending on the receptor being evaluated.
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Adulto , Anciano , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Quimiocinas/sangre , Integrinas/sangre , Monocitos/química , Neutrófilos/química , Sepsis/inmunología , Receptores Toll-Like/sangre , Antibacterianos/uso terapéutico , Antígenos CD/sangre , /sangre , /sangre , Moléculas de Adhesión Celular/sangre , Citometría de Flujo , Proteínas Ligadas a GPI/sangre , Mortalidad Hospitalaria , Inmunofenotipificación , Unidades de Cuidados Intensivos , /sangre , Estadísticas no Paramétricas , Sepsis/terapia , Resultado del Tratamiento , Receptor Toll-Like 9/sangre , /sangre , /sangre , /sangreRESUMEN
Recognition of pathogens is performed by specific receptors in cells of the innate immune system, which may undergo modulation during the continuum of clinical manifestations of sepsis. Monocytes and neutrophils play a key role in host defense by sensing and destroying microorganisms. This study aimed to evaluate the expression of CD14 receptors on monocytes; CD66b and CXCR2 receptors on neutrophils; and TLR2, TLR4, TLR5, TLR9, and CD11b receptors on both cell types of septic patients. Seventy-seven septic patients (SP) and 40 healthy volunteers (HV) were included in the study, and blood samples were collected on day zero (D0) and after 7 days of therapy (D7). Evaluation of the cellular receptors was carried out by flow cytometry. Expression of CD14 on monocytes and of CD11b and CXCR2 on neutrophils from SP was lower than that from HV. Conversely, expression of TLR5 on monocytes and neutrophils was higher in SP compared with HV. Expression of TLR2 on the surface of neutrophils and that of TLR5 on monocytes and neutrophils of SP was lower at D7 than at D0. In addition, SP who survived showed reduced expression of TLR2 and TLR4 on the surface of neutrophils at D7 compared to D0. Expression of CXCR2 for surviving patients was higher at follow-up compared to baseline. We conclude that expression of recognition and cell signaling receptors is differentially regulated between SP and HV depending on the receptor being evaluated.
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Quimiocinas/sangre , Integrinas/sangre , Monocitos/química , Neutrófilos/química , Sepsis/inmunología , Receptores Toll-Like/sangre , Adulto , Anciano , Antibacterianos/uso terapéutico , Antígenos CD/sangre , Antígeno CD11b/sangre , Moléculas de Adhesión Celular/sangre , Preescolar , Femenino , Citometría de Flujo , Proteínas Ligadas a GPI/sangre , Mortalidad Hospitalaria , Humanos , Inmunofenotipificación , Unidades de Cuidados Intensivos , Receptores de Lipopolisacáridos/sangre , Masculino , Persona de Mediana Edad , Receptores de Interleucina-8B/sangre , Sepsis/terapia , Estadísticas no Paramétricas , Receptor Toll-Like 2/sangre , Receptor Toll-Like 4/sangre , Receptor Toll-Like 5/sangre , Receptor Toll-Like 9/sangre , Resultado del TratamientoRESUMEN
Lipopolysaccharide stimulation of toll-like receptor 4 (TLR4) activates signal transduction pathways leading to proinflammatory cytokine secretion. We investigated TLR4 surface receptor expression on peripheral blood neutrophils and monocytes and their ability to modulate inflammatory cytokine release in 15 patients 1, 3, and 10 days after hemolytic uremic syndrome (HUS) onset. Seven patients with Escherichia coli (EHEC)-associated diarrhea and seven healthy controls were also studied. Isolated leucocytes from HUS-onset patients exhibited significantly higher messenger RNA (mRNA) TLR4 expression than controls. Moreover, TLR4 protein expression on neutrophils, determined by flow cytometry, was upregulated, driving dependent proinflammatory cytokine, tumor necrosis factor alpha (TNF-α), and interleukin 8 (IL-8) increase, and decreased anti-inflammatory IL-10 release at HUS onset compared with patients with EHEC diarrhea and controls. TLR4 expression on neutrophils was positively correlated with serum TNF-α levels. Conversely, significant reduction of neutrophil TLR4 receptor expression and lack of cytokine-responsive element activation was shown in patients 3 and 10 days after HUS onset. No differences were demonstrated in TLR4 receptor expression on monocytes among the studied groups. Our results suggest TLR4 expression may be differently regulated on neutrophils and monocytes. They could be dynamically modulated across the early development of HUS on neutrophils, resulting in negative regulation preceded by TLR4 overactivation.
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Síndrome Hemolítico-Urémico/metabolismo , Leucocitos/metabolismo , Receptor Toll-Like 4/genética , Preescolar , Citocinas/sangre , Femenino , Regulación de la Expresión Génica , Humanos , Lactante , Masculino , Monocitos/química , Neutrófilos/química , Receptor Toll-Like 4/sangreRESUMEN
BACKGROUND: Intravascular hemolysis may cause tissue injury directly or via a systemic inï¬ammatory response. Under physiological conditions, extracorpuscular hemoglobin (Hb) is bound by haptoglobin(Hp) and the complex internalized via the hemoglobin scavenger receptor CD163 on monocytes, prior to catabolism via heme-oxygenase-1 (HO-1). Recently, a novel subset of CD68(pos)CD163(high)HLA-DR(low) macrophages with high expression of HO-1 was recognized in hemorrhagic areas of atherosclerotic plaques, distinct from CD68(pos)CD163(low)HLA-DR(high) foam cell macrophages with low- HO-1 content. Considering the hemolytic insult during CPB, we hypothesized that an equivalent compensatory CD163(high)HLA-DR(low) phenotype will evolve in circulating CD14(pos) monocytes post surgery. METHODS: Twelve patients undergoing elective surgery with CPB were enrolled with informed consent.Whole-blood samples were collected in EDTA at predetermined time-points, pre- intra-, and postoperatively. Whole-blood was evaluated by three-color flow cytometry for expression of CD14, CD163, and HLA-DR; CD14(pos) cells were also permeabilized to detect intracellular HO-1 protein. Plasma [Hp-Hb] concentration was determined by ELISA. RESULTS: A striking phenotypic switch from CD163(low)HLA-DR(high) preoperatively to CD163(high)HLA-DR(low) postoperatively at 24 h was observed on circulating CD14(pos) monocytes. Intracellular HO-1 protein was also significantly up-regulated at 24 h after declamping. These phenotypic changes were preceded by peak Hb-Hp levels observed at 2 h. CONCLUSION: We have shown for the first time, a phenotypic commitment of monocytes towards a protective CD14(pos)CD163(high)HLA-DR(low) population with increased intracellular HO-1 occurring in the circulation during the recovery phase of CPB. These findings have implications for monitoring of anti-inflammatory interventions and linkage to clinical outcomes.
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Puente Cardiopulmonar , Hemoglobinas/metabolismo , Monocitos/metabolismo , Antígenos CD/sangre , Antígenos CD/inmunología , Antígenos de Diferenciación Mielomonocítica/sangre , Antígenos de Diferenciación Mielomonocítica/inmunología , Citometría de Flujo , Células Espumosas/química , Células Espumosas/inmunología , Células Espumosas/metabolismo , Antígenos HLA-DR/sangre , Antígenos HLA-DR/inmunología , Haptoglobinas/análisis , Haptoglobinas/inmunología , Hemo-Oxigenasa 1/análisis , Humanos , Receptores de Lipopolisacáridos/sangre , Receptores de Lipopolisacáridos/inmunología , Monocitos/química , Monocitos/inmunología , Fenotipo , Periodo Posoperatorio , Receptores de Superficie Celular/sangre , Receptores de Superficie Celular/inmunología , Regulación hacia ArribaRESUMEN
Yellow fever (YF) vaccines (17D-204 and 17DD) are well tolerated and cause very low rates of severe adverse events (YEL-SAE), such as serious allergic reactions, neurotropic adverse diseases (YEL-AND), and viscerotropic diseases (YEL-AVD). Viral and host factors have been postulated to explain the basis of YEL-SAE. However, the mechanisms underlying the occurrence of YEL-SAE remain unknown. The present report provides a detailed immunological analysis of a 23-year-old female patient. The patient developed a suspected case of severe YEL-AVD with encephalitis, as well as with pancreatitis and myositis, following receipt of a 17D-204 YF vaccination. The patient exhibited a decreased level of expression of Fc-gammaR in monocytes (CD16, CD32, and CD64), along with increased levels of NK T cells (an increased CD3(+) CD16(+/-) CD56(+/-)/CD3(+) ratio), activated T cells (CD4(+) and CD8(+) cells), and B lymphocytes. Enhanced levels of plasmatic cytokines (interleukin-6 [IL-6], IL-17, IL-4, IL-5, and IL-10) as well as an exacerbated ex vivo intracytoplasmic cytokine pattern, mainly observed within NK cells (gamma interferon positive [IFN-gamma(+)], tumor necrosis factor alpha positive [TNF-alpha(+)], and IL-4 positive [IL-4(+)]), CD8(+) T cells (IL-4(+) and IL-5(+)), and B lymphocytes (TNF-alpha(+), IL-4(+), and IL-10(+)). The analysis of CD4(+) T cells revealed a complex profile that consisted of an increased frequency of IL-12(+) and IFN-gamma(+) cells and a decreased percentage of TNF-alpha(+), IL-4(+), and IL-5(+) cells. Depressed cytokine synthesis was observed in monocytes (TNF-alpha(+)) following the provision of antigenic stimuli in vitro. These results support the hypothesis that a strong adaptive response and abnormalities in the innate immune system may be involved in the establishment of YEL-AND and YEL-AVD.
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Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/inmunología , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/patología , Vacuna contra la Fiebre Amarilla/efectos adversos , Vacuna contra la Fiebre Amarilla/inmunología , Linfocitos B/inmunología , Citocinas/biosíntesis , Citocinas/sangre , Encefalitis/inducido químicamente , Encefalitis/complicaciones , Encefalitis/patología , Femenino , Humanos , Células Asesinas Naturales/química , Células Asesinas Naturales/inmunología , Monocitos/química , Monocitos/inmunología , Miositis/inducido químicamente , Miositis/complicaciones , Miositis/patología , Pancreatitis/inducido químicamente , Pancreatitis/complicaciones , Pancreatitis/patología , Linfocitos T/química , Linfocitos T/inmunología , Adulto JovenRESUMEN
INTRODUCTION: Tissue factor (TF) is the main physiological initiator of blood coagulation; it is membrane-bound on monocytes (mTF) and free in plasma (pTF). Abnormal expression of TF by monocytes has been implicated in various diseases. We therefore quantified monocytes expressing TF and pTF levels in patients with lower-limb deep venous thrombosis (DVT). MATERIALS AND METHODS: DVT was confirmed by Duplex Scan. Blood mTF levels under resting condition (baseline), after incubation without (unstimulated) and with (stimulated) lipopolysaccharide (LPS), and total mTF levels were determined by flow cytometry using two analytical methods (Histogram and Quadrant-Statistics). Plasma TF levels were measured using an enzyme-linked immunoabsorbent assay (ELISA). Results were compared with age-matched controls. RESULTS: Histogram analysis in patients with DVT showed significantly elevated mTF levels for baseline, unstimulated and total mTF over controls. For Quadrant-Statistics, DVT patients also showed significantly raised baseline, unstimulated, stimulated and total mTF. Similarly, pTF levels were significantly raised in subjects with DVT compared to controls. Baseline mTF levels correlated with pTF levels by Histogram and Quadrant-Statistics analysis. Using the relative operating characteristic (ROC) curve, baseline mTF and pTF assays displayed sensitivity and specificity in detecting DVT. Quadrant-Statistics baseline mTF and pTF gave the best discrimination. CONCLUSIONS: The TF assays used in this study showed acceptable sensitivity and specificity and are cost-effective and practical. Therefore, they should be considered in patients with, or at risk of, DVT.
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Monocitos/química , Plasma/química , Tromboplastina/análisis , Trombosis de la Vena/diagnóstico , Adulto , Anciano , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Humanos , Lipopolisacáridos/farmacología , Extremidad Inferior/irrigación sanguínea , Masculino , Persona de Mediana Edad , Monocitos/citología , Sensibilidad y EspecificidadRESUMEN
Dendritic cells (DC) play a dual role in the immune response, participating in its induction, and the maintenance of immune tolerance. The aim of this work was to perform a quantitative and phenotypic analysis of DC generated in vitro in the presence of IL-10 in patients with systemic lupus erythematosus (SLE). Blood samples were obtained from 10 active and untreated patients with SLE and six controls. Monocyte-derived DC were generated in vitro in the presence or absence of IL-10, and a quantitative and phenotypic analysis was performed. We found that freshly isolated monocytes from SLE patients had an increased expression of CD11b. On the other hand, the efficiency of in vitro DC generation was diminished in blood samples from SLE patients for conventional DC, but not for IL-10-treated DC. A diminished expression of HLA-DR, CD9 and CD86 was observed in conventional DC from SLE patients compared with controls. In contrast, enhanced levels of HLA-DR, CD80, CD9 and CD151 tetraspanins, FN1 (a class II MHC-tetraspanin epitope), CD85j/ILT2 and CD69 were detected in IL-10-treated DC from SLE patients. Accordingly, the phenotypic profile of IL-10-treated DC was very different in SLE and controls. However, the synthesis of IL-10 and IL-12 was similar in IL-10-treated and conventional cells in both SLE patients and controls. Our findings on the aberrant phenotype of IL-10-treated DC in SLE and their normal efficiency of in vitro generation may be important for the design of future therapies of this condition based on the administration of DC to induce immune tolerance.
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Antígenos CD/análisis , Células Dendríticas/química , Células Dendríticas/efectos de los fármacos , Antígenos HLA-DR/análisis , Interleucina-10/farmacología , Lupus Eritematoso Sistémico/inmunología , Adulto , Diferenciación Celular , Células Dendríticas/inmunología , Femenino , Citometría de Flujo , Humanos , Masculino , Persona de Mediana Edad , Monocitos/química , Monocitos/citología , Fenotipo , Receptores de Superficie Celular/análisisRESUMEN
Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of 'piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.
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Plaquetas/ultraestructura , Peces/fisiología , Leucocitos/ultraestructura , Animales , Biomarcadores/análisis , Plaquetas/química , Gránulos Citoplasmáticos/química , Técnicas para Inmunoenzimas , Leucocitos/química , Microscopía Electrónica de Transmisión/métodos , Monocitos/química , Monocitos/ultraestructura , Neutrófilos/química , Neutrófilos/ultraestructura , Reacción del Ácido Peryódico de Schiff/métodosRESUMEN
The photosensitizing properties of six anthraquinones (AQs): soranjidiol (1), soranjidiol-1-methyl ether (2), rubiadin (3), rubiadin-1-methyl ether (4), damnacanthal (5) and damnacanthol (6), isolated from leaves and stems of Heterophyllaea pustulata Hook. f. (Rubiaceae) were studied. By means of photobiological and photophysical methods in vitro, the type of photosensitization that these metabolites are capable of producing was determined. Whereas the photosensitized generation of superoxide anion radical (O(2)(-)) (Type I) was evaluated in leukocyte suspensions, singlet molecular oxygen ((1)O(2)) production (Type II) was examined in organic solution. In addition, the quantum yield of (1)O(2) (Phi) in chloroform was measured for those AQs that generate it. It was established that 4 behaves exclusively as a Type I photosensitizer. By contrast, the others AQs act by both types of mechanisms, among which 5 showed the largest Phi of (1)O(2).
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Antraquinonas/química , Fármacos Fotosensibilizantes/química , Oxígeno Singlete/química , Oxígeno Singlete/metabolismo , Superóxidos/química , Superóxidos/metabolismo , Aniones/química , Humanos , Estructura Molecular , Monocitos/química , Monocitos/metabolismo , Análisis EspectralRESUMEN
The localization of peroxidase activity in different cell regions is used as a criterion for classifying the stage of maturity of mammalian mononuclear phagocytes, with a positive peroxidase reaction indicating the presence of monoblasts, promonocytes, monocytes, and macrophages. Peroxidase activity was observed ultrastructurally in the circulating blood of pacu fish (Piaractus mesopotamicus), identifying monoblasts, promonocytes, monocytes, and macrophages. These observations suggest that differentiation of mononuclear phagocytes occurs in the blood circulation of fish, whereas in mammals, monoblasts and promonocytes are detected in bone marrow, with only monocytes detected in circulating blood and differentiation into macrophages occurring in other body compartments.