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1.
EBioMedicine ; 59: 102964, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32861199

RESUMO

Mononuclear phagocytes are a widely distributed family of cells contributing to innate and adaptive immunity. Circulating monocytes and tissue macrophages participate in all stages of SARS COVID-19. They contribute to comorbidities predisposing to clinical infection, virus resistance and dissemination, and to host factors that determine disease severity, recovery and sequelae. Assays are available to detect viral infection and antibody responses, but no adequate tests have been developed to measure the activation level of monocytes and tissue macrophages, and the risk of progression to a fatal hyperinflammatory syndrome. Blood monocytes provide a window on the systemic immune response, from production to tissue recruitment, reflecting the impact of infection on the host. Ready availability of blood makes it possible to monitor severity and the risk of potentially lethal complications, by developing tests to assess the status of monocyte activation and its potential for further inflammatory dysregulation after recruitment to tissues and during recovery.


Assuntos
Infecções por Coronavirus/patologia , Monócitos/imunologia , Pneumonia Viral/patologia , Betacoronavirus/isolamento & purificação , Comorbidade , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Citocinas/metabolismo , Humanos , Imunidade Inata , Macrófagos/citologia , Macrófagos/imunologia , Macrófagos/metabolismo , Monócitos/citologia , Monócitos/metabolismo , Pandemias , Pneumonia Viral/imunologia , Pneumonia Viral/virologia , Índice de Gravidade de Doença
2.
Proc Natl Acad Sci U S A ; 117(26): 14798-14804, 2020 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-32554496

RESUMO

Proper circulation of white blood cells (WBCs) in the pulmonary vascular bed is crucial for an effective immune response. In this branched vascular network, WBCs have to strongly deform to pass through the narrowest capillaries and bifurcations. Although it is known that this process depends on the cell mechanical properties, it is still poorly understood due to the lack of a comprehensive model of cell mechanics and of physiologically relevant experiments. Here, using an in-house microfluidic device mimicking the pulmonary capillary bed, we show that the dynamics of THP1 monocytes evolves along successive capillary-like channels, from a nonstationary slow motion with hops to a fast and smooth efficient one. We used actin cytoskeleton drugs to modify the traffic dynamics. This led us to propose a simple mechanical model that shows that a very finely tuned cortical tension combined with a high cell viscosity governs the fast transit through the network while preserving cell integrity. We finally highlight that the cortical tension controls the steady-state cell velocity via the viscous friction between the cell and the channel walls.


Assuntos
Capilares/fisiologia , Pulmão , Modelos Biológicos , Monócitos , Fenômenos Biomecânicos , Humanos , Pulmão/irrigação sanguínea , Pulmão/citologia , Técnicas Analíticas Microfluídicas/instrumentação , Monócitos/citologia , Monócitos/fisiologia , Células THP-1
3.
PLoS One ; 15(6): e0234038, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32492075

RESUMO

Extracellular adenosine triphosphate (eATP) released by damaged cells, and its purinergic receptors, comprise a crucial signaling network after injury. Purinergic receptor P2X7 (P2RX7), a major driver of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activation and IL-1ß processing, has been shown to play a role in liver injury in murine diet- and chemically-induced liver injury models. It is unclear, however, whether P2RX7 plays a role in non-alcoholic steatohepatitis (NASH) and which cell type is the main target of P2RX7 pharmacological inhibition. Here, we report that P2RX7 is expressed by infiltrating monocytes and resident Kupffer cells in livers from NASH-affected individuals. Using primary isolated human cells, we demonstrate that P2RX7 expression in CD14+ monocytes and Kupffer cells primarily mediates IL-1ß release. In addition, we show that pharmacological inhibition of P2RX7 in monocytes and Kupffer cells, blocks IL-1ß release, reducing hepatocyte caspase 3/7 activity, IL-1ß-mediated CCL2 and CCL5 chemokine gene expression and secretion, and hepatic stellate cell (HSC) procollagen secretion. Consequently, in a chemically-induced nonhuman primate model of liver fibrosis, treatment with a P2RX7 inhibitor improved histological characteristics of NASH, protecting from liver inflammation and fibrosis. Taken together, these findings underscore the critical role of P2RX7 in the pathogenesis of NASH and implicate P2RX7 as a promising therapeutic target for the management of this disease.


Assuntos
Inflamação/prevenção & controle , Cirrose Hepática/tratamento farmacológico , Antagonistas do Receptor Purinérgico P2X/uso terapêutico , Receptores Purinérgicos P2X7/metabolismo , Animais , Caspase 3/metabolismo , Células Cultivadas , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Células Estreladas do Fígado/citologia , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Inflamação/patologia , Interleucina-1beta/metabolismo , Macrófagos do Fígado/citologia , Macrófagos do Fígado/efeitos dos fármacos , Macrófagos do Fígado/metabolismo , Lipopolissacarídeos/farmacologia , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/patologia , Macaca fascicularis , Masculino , Monócitos/citologia , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Pró-Colágeno/metabolismo , Antagonistas do Receptor Purinérgico P2X/farmacologia , Receptores Purinérgicos P2X7/química , Receptores Purinérgicos P2X7/genética
4.
PLoS One ; 15(6): e0235174, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32574226

RESUMO

AIM: To investigate markers of systemic inflammation in pre- and postmenopausal women and identify possible predictors of systemic inflammation with menopause. METHODS: Cross-sectional study of 69 healthy women between 45- and 60 years. Blood samples were collected to assess leukocyte subsets and plasma cytokines. MRI and DXA scans were performed to assess body composition. Through uni- and multivariate analyses, follicle-stimulating hormone (FSH), visceral fat mass and age were evaluated as predictors of systemic inflammation in relation to menopause. RESULTS: Postmenopausal women tended to have higher leukocyte counts (5.4 x109 vs. 4.9 x109 cells/l, p = 0.05) reflected in increased total lymphocytes (1.8 x109 vs. 1.6 x109 cells/l, p = 0.01) and monocytes (0.5 x109 vs. 0.4 x109 cells/l, p = 0.02), compared to premenopausal women. Increased visceral fat mass was a strong predictor of high leukocyte subsets. Postmenopausal women had higher plasma TNF-α (2.24 vs. 1.91 pg/ml, p = 0.01) and IL-6 (0.45 vs. 0.33 pg/ml, p = 0.004) compared to premenopausal women and high FSH was a significant predictor of increased plasma TNF-α, IL-1ß and IL-6. Menopause was further associated with increased T-cells (1,336 vs. 1,128 cells/µl, p = 0.04) reflected in significantly higher counts of exhausted-, senescent-, and memory CD4+ T-cell subsets. CONCLUSIONS: Menopause is associated with increased systemic inflammation as well as exhausted- and senescent T-cells. We suggest, that both increased visceral fat mass and declining sex hormone levels might contribute to postmenopausal systemic inflammation and calls for further large-scale studies to confirm these findings.


Assuntos
Citocinas/imunologia , Inflamação/imunologia , Pós-Menopausa/imunologia , Subpopulações de Linfócitos T/imunologia , Absorciometria de Fóton/métodos , Composição Corporal , Citocinas/sangue , Citocinas/metabolismo , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Foliculoestimulante/imunologia , Hormônios Esteroides Gonadais/sangue , Hormônios Esteroides Gonadais/imunologia , Humanos , Inflamação/sangue , Inflamação/metabolismo , Gordura Intra-Abdominal/diagnóstico por imagem , Gordura Intra-Abdominal/imunologia , Gordura Intra-Abdominal/metabolismo , Linfócitos/citologia , Linfócitos/imunologia , Linfócitos/metabolismo , Imagem por Ressonância Magnética/métodos , Pessoa de Meia-Idade , Monócitos/citologia , Monócitos/imunologia , Monócitos/metabolismo , Análise Multivariada , Pós-Menopausa/sangue , Pós-Menopausa/metabolismo , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/metabolismo
5.
Cells ; 9(6)2020 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-32526950

RESUMO

Feline coronavirus is a highly contagious virus potentially resulting in feline infectious peritonitis (FIP), while the pathogenesis of FIP remains not well understood, particularly in the events leading to the disease. A predominant theory is that the pathogenic FIPV arises from a mutation, so that it could replicate not only in enterocytes of the intestines but also in monocytes, subsequently systemically transporting the virus. The immune status and genetics of affected cats certainly play an important role in the pathogenesis. Considering the importance of genetics and host immune responses in viral infections, the goal of this study was to elucidate host gene expression in macrophages using RNA sequencing. Macrophages from healthy male cats infected with FIPV 79-1146 ex vivo displayed a differential host gene expression. Despite the virus uptake, aligned viral reads did not increase from 2 to 17 h. The overlap of host gene expression among macrophages from different cats was limited, even though viral transcripts were detected in the cells. Interestingly, some of the downregulated genes in all macrophages were involved in immune signaling, while some upregulated genes common for all cats were found to be inhibiting immune activation. Our results highlight individual host responses playing an important role, consistent with the fact that few cats develop feline infectious peritonitis despite a common presence of enteric FCoV.


Assuntos
Coronavirus Felino/imunologia , Peritonite Infecciosa Felina/imunologia , Peritonite Infecciosa Felina/patologia , Macrófagos/imunologia , Monócitos/imunologia , Animais , Gatos , Linhagem Celular , Coronavirus Felino/genética , Peritonite Infecciosa Felina/virologia , Regulação da Expressão Gênica , Imunidade Inata/genética , Imunidade Inata/imunologia , Macrófagos/citologia , Monócitos/citologia , RNA Viral/isolamento & purificação , Análise de Sequência de RNA , Transcriptoma/genética
6.
Medicine (Baltimore) ; 99(19): e20190, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32384513

RESUMO

Diabetic nephropathy (DN) is serious threat to human health. Therefore, early prediction of its occurrence is important. This study aimed to assess the predictive significance of monocyte-lymphocyte ratio (MLR) for DN.A total of 301 patients with type 2 diabetes (T2D), including 212 T2D patients without diabetic-related complications and 99 DN patients, were enrolled. Peripheral white blood cells were measured before treatment to calculate MLR, and the risk factors and predictive significance for T2D and DN were assessed.T2D patients without diabetic-related complications had higher MLR than control patients (P < .01). However, MLR was significantly higher in DN patients than in T2D patients without diabetic-related complications (P < .001). According to MLR quartiles, higher MLR in DN patients was correlated with higher serum creatinine, estimated glomerular filtration rate, and urinary albumin excretion (UAE) levels (P < .01 or P < .001). Furthermore, MLR was positively correlated with UAE level (R = 0.5973; P < .01) and an independent predictor for DN (odds ratio: 7.667; 95% confidence interval [CI]: 3.689-21.312; P < .001). The area under the receiver-operating characteristic (ROC) curve for MLR was 0.874 (95%CI: 0.830-0.918, P < .001). When the optimal cutoff value was 0.23, the sensitivity and specificity of MLR for DN prediction were 0.85 and 0.74, respectively.The present findings suggest that MLR is a powerful independent predictor for DN.


Assuntos
Diabetes Mellitus Tipo 2/sangue , Nefropatias Diabéticas/sangue , Linfócitos/citologia , Monócitos/citologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Albuminúria , Creatinina/sangue , Feminino , Taxa de Filtração Glomerular , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Curva ROC , Fatores de Risco
7.
Bratisl Lek Listy ; 121(4): 287-292, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32356444

RESUMO

OBJECTIVES: The aim of our study was to assess the diagnostic value of neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), platelet-to-lymphocyte ratio (PLR), and red blood cell distribution width (RDW) in children with lymphadenopathy (LAP). METHODS: Between January, 2009 and December, 2018, 190 children who underwent excisional lymph node biopsy due to enlarged lymph nodes were included. The clinical and laboratory features of pediatric patients with lymph node enlargement, histopathological examination of the lymph node, and the role of complete blood count parameters in the differentiation of reactive and malignant LAP were analyzed retrospectively. RESULTS: In total, 139 (73.2 %) children had pathologically confirmed reactive LAP and 51 (26.8 %) were diagnosed with lymphoma. Compared with the reactive LAP group, median values for NLR, MLR, PLR, and RDW were significantly higher in children with lymphoma (p < 0.01). According to receiver operating characteristic curve analysis performed for distinguishing between reactive LAP and lymphomas, the area under curves of NLR, MLR, PLR, and RDW were 0.75, 0.76, 0.71, and 0.61, respectively. CONCLUSIONS: Children with histologically proven lymphoma have higher NLR, MLR, PLR, and RDW values than children with reactive LAP. NLR, MLR, PLR, and RDW tests, which can be performed even in primary health care centers, may be useful markers to determine which patients with LAP should be referred to the advanced center at an early stage for biopsy (Tab. 4, Fig. 2, Ref. 26).


Assuntos
Plaquetas/citologia , Índices de Eritrócitos , Linfadenopatia/diagnóstico , Linfócitos/citologia , Linfoma/diagnóstico , Monócitos/citologia , Neutrófilos/citologia , Criança , Humanos , Linfadenopatia/sangue , Linfoma/sangue , Prognóstico , Estudos Retrospectivos
8.
Mol Immunol ; 123: 1-6, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32380279

RESUMO

The repertoire of dendritic cells (DCs), monocytes and macrophages in adult humans is diverse and we are appreciating this to a greater extent as high throughput methods, such a single-cell RNA sequencing, become widely adopted and scalable. This powerful lens of analysis is also beginning to shed light on prenatal immunology, allowing us to chart the emergence, tissue distribution and developmental regulation of DCs, monocytes and macrophages during early human life. In this review, we will integrate recent insights from studies of the developing immune system into our understanding of adult DC, monocyte and macrophage organization, illustrating where insights from early life both affirm and challenge current understanding.


Assuntos
Células Dendríticas/citologia , Desenvolvimento Fetal/fisiologia , Macrófagos/citologia , Monócitos/citologia , Mielopoese/fisiologia , Análise de Célula Única/métodos , Adulto , Animais , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Células Cultivadas , Células Dendríticas/fisiologia , Feminino , Humanos , Macrófagos/fisiologia , Monócitos/fisiologia , Gravidez
9.
Pol J Pathol ; 71(1): 46-54, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32429654

RESUMO

Prostate cancer is one of the most frequent cancers in men. Although several treatment options exist, their clinical effectiveness is still not satisfactory. One the possible reason of such situation might be the presence of myeloid-derived suppressor cells (MDSC) and their pro-tumorigenic activity. MDSC possess immunosuppressive ability and in many studies were shown to support tumor development and progression. In this study we addressed the question whether commonly used therapies of prostate cancer affect the level of MDSC populations in the patients' blood. We compared the level of granulocytic (Gr-MDSC), monocytic (Mo-MDSC) and early stage MDSC (eMDSC) in the blood of patients at different clinical stage and different tumor grading scores, who underwent either surgery or hormonal therapy alone or were given a combined treatment, including e.g. radiotherapy. The obtained results showed that the level of Gr-MDSC was significantly lower in all treated patients comparing to untreated group. On the other hand, surgery or hormonal therapy alone did not affect the level of Mo-MDSC. These results were independent of the PSA level, the tumor grading and clinical stage of the patients. In conclusion, we suggest that Mo-MDSC should be considered as a potential therapy target in the course of prostate cancer treatment to enhance its anti-tumor effectiveness.


Assuntos
Células Supressoras Mieloides/citologia , Neoplasias da Próstata/sangue , Granulócitos/citologia , Humanos , Masculino , Monócitos/citologia , Neoplasias da Próstata/terapia , Resultado do Tratamento
10.
Nat Commun ; 11(1): 2280, 2020 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-32385245

RESUMO

Renal macrophages (RMs) participate in tissue homeostasis, inflammation and repair. RMs consist of embryo-derived (EMRMs) and bone marrow-derived RMs (BMRMs), but the fate, dynamics, replenishment, functions and metabolic states of these two RM populations remain unclear. Here we investigate and characterize RMs at different ages by conditionally labeling and ablating RMs populations in several transgenic lines. We find that RMs expand and mature in parallel with renal growth after birth, and are mainly derived from fetal liver monocytes before birth, but self-maintain through adulthood with contribution from peripheral monocytes. Moreover, after the RMs niche is emptied, peripheral monocytes rapidly differentiate into BMRMs, with the CX3CR1/CX3CL1 signaling axis being essential for the maintenance and regeneration of both EMRMs and BMRMs. Lastly, we show that EMRMs have a higher capacity for scavenging immune complex, and are more sensitive to immune challenge than BMRMs, with this difference associated with their distinct glycolytic capacities.


Assuntos
Células da Medula Óssea/citologia , Linhagem da Célula , Rim/embriologia , Macrófagos/citologia , Animais , Receptor 1 de Quimiocina CX3C/metabolismo , Quimiocina CX3CL1/sangue , Quimiocina CX3CL1/metabolismo , Feminino , Feto/citologia , Fígado/embriologia , Masculino , Camundongos , Monócitos/citologia
11.
J Vis Exp ; (159)2020 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-32449722

RESUMO

A human alveolar cell coculture model is described here for simulation of the alveolar epithelial tissue barrier composed of alveolar epithelial type II cells and two types of immune cells (i.e., human monocyte-derived macrophages [MDMs] and dendritic cells [MDDCs]). A protocol for assembling the multicellular model is provided. Alveolar epithelial cells (A549 cell line) are grown and differentiated under submerged conditions on permeable inserts in two-chamber wells, then combined with differentiated MDMs and MDDCs. Finally, the cells are exposed to an air-liquid interface for several days. As human primary immune cells need to be isolated from human buffy coats, immune cells differentiated from either fresh or thawed monocytes are compared in order to tailor the method based on experimental needs. The three-dimensional models, composed of alveolar cells with either freshly isolated or thawed monocyte-derived immune cells, show a statistically significant increase in cytokine (interleukins 6 and 8) release upon exposure to proinflammatory stimuli (lipopolysaccharide and tumor necrosis factor α) compared to untreated cells. On the other hand, there is no statistically significant difference between the cytokine release observed in the cocultures. This shows that the presented model is responsive to proinflammatory stimuli in the presence of MDMs and MDDCs differentiated from fresh or thawed peripheral blood monocytes (PBMs). Thus, it is a powerful tool for investigations of acute biological response to different substances, including aerosolized drugs or nanomaterials.


Assuntos
Células Epiteliais Alveolares/citologia , Células Epiteliais/citologia , Macrófagos/citologia , Modelos Biológicos , Células A549 , Células Epiteliais Alveolares/metabolismo , Contagem de Células , Morte Celular , Diferenciação Celular , Proliferação de Células , Forma Celular , Técnicas de Cocultura , Congelamento , Humanos , Mediadores da Inflamação/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Monócitos/citologia
12.
PLoS One ; 15(4): e0231927, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32343751

RESUMO

Metabolic Syndrome (MS) is characterized by a low-grade inflammatory state causing an alteration of non-invasive indexes derived from blood count, namely monocyte-to-HDL ratio (MHR), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR). We analyse a population of 771 subjects (394 controls and 377 MS patients) to evaluate the best predictive index of MS. The diagnosis of MS was made according to the 2006 criteria of the International Diabetes Federation (IDF). We performed ROC curve analyses to evaluate the best predictor index of MS. MHR cut-off value was used to classify the population in two different groups and to create the outcome variable of the Recursive Partitioning and Amalgamation (RECPAM) analysis. This method is a tree-structured approach that defines "risk profiles" for each group of dichotomous variables. We showed that MHR index is significantly linked to body mass index (BMI), waist circumference, creatinine, C-reactive protein (CRP), Erythrocyte Sedimentation Rate (ESR). ROC curve defined an MHR cut-off value of 6.4, which was able to identify two patient groups with significant differences in waist circumference, blood pressure, creatinine, estimated glomerular filtration rate and fasting plasma glucose. RECPAM analysis demonstrated that gender, BMI categorization and hyperglycaemia were the most important risk determinants of increased MHR index that can be considered bona fide a useful and easily obtainable tool to suggest the presence of peculiar metabolic features that predict MS.


Assuntos
Glicemia/análise , Índice de Massa Corporal , Lipoproteínas HDL/sangue , Síndrome Metabólica/patologia , Monócitos/citologia , Adulto , Idoso , Área Sob a Curva , Sedimentação Sanguínea , Proteína C-Reativa/análise , Estudos de Casos e Controles , Feminino , Humanos , Modelos Logísticos , Masculino , Síndrome Metabólica/sangue , Síndrome Metabólica/metabolismo , Pessoa de Meia-Idade , Curva ROC , Fatores Sexuais , Fumantes , Circunferência da Cintura
13.
Scand J Immunol ; 92(1): e12883, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32243617

RESUMO

Monocytes are important cells of the innate system. They are a heterogeneous type of cells consisting of phenotypically and functionally distinct subpopulations, which play a specific role in the control, development and escalation of the immunological processes. Based on the expression of superficial CD14 and CD16 in flow cytometry, they can be divided into three subsets: classical, intermediate and non-classical. Variation in the levels of human monocyte subsets in the blood can be observed in patients in numerous pathological states, such as infections, cardiovascular and inflammatory diseases, cancer and autoimmune diseases. The aim of this review is to summarize current knowledge of human monocyte subsets and their significance in homeostasis and in pathological conditions.


Assuntos
Imunidade Inata/imunologia , Monócitos/classificação , Monócitos/imunologia , Fatores Estimuladores de Colônias/biossíntese , Fatores Estimuladores de Colônias/imunologia , Humanos , Macrófagos/citologia , Macrófagos/imunologia , Monócitos/citologia , Receptores de Superfície Celular/imunologia
14.
Am J Med Sci ; 359(5): 281-286, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32245567

RESUMO

BACKGROUND: The monocyte to high-density lipoprotein ratio (MHR) has been used to predict adverse clinical outcomes in patients with acute coronary syndrome (ACS). This meta-analysis aimed to evaluate the prognostic utility of MHR in patients with ACS. MATERIALS AND METHODS: We comprehensively searched for relevant studies in Pubmed, Embase, CNKI, WanFang and VIP databases until March 12, 2019. Epidemiologic studies investigating the association between MHR and major adverse cardiovascular events (MACE) or all-cause mortality in patients with ACS were included. Pooled effect was expressed as risk ratios (RR) with 95% confidence intervals (CI) for the highest versus the reference lower MHR group. RESULTS: Eight studies involving 6,480 patients with ACS were included and analyzed. Meta-analysis indicated that the highest MHR was significantly associated with higher risk of MACE (RR 1.65; 95%CI 1.36-2.02) and all-cause mortality (RR 2.61; 95%CI 1.29-4.89) after adjusting for the conventional confounders. The prognostic values of MACE with the highest MHR caused no significant changes in the in-hospital follow-up (RR 1.76; 95%CI 1.34-2.32) and >6 months follow-up (RR 1.68; 95%CI 1.08-2.62) subgroups. Furthermore, ST elevation myocardial infarction patients with the highest MHR had a 2.07-fold higher risk of in-hospital MACE (RR 2.07; 95%CI 1.52-2.80). CONCLUSIONS: Elevated MHR is independently associated with an increased risk of MACE and all-cause mortality in patients with ACS. MHR may serve as a potential prognostic indicator for ACS prognosis.


Assuntos
Síndrome Coronariana Aguda/sangue , Síndrome Coronariana Aguda/diagnóstico , Lipoproteínas HDL/sangue , Monócitos/citologia , Feminino , Humanos , Inflamação , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo , Prognóstico , Fatores de Risco , Infarto do Miocárdio com Supradesnível do Segmento ST/sangue , Infarto do Miocárdio com Supradesnível do Segmento ST/diagnóstico
15.
PLoS One ; 15(4): e0231132, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32271804

RESUMO

The monocyte-derived dendritic cells (moDCs) are a subset of dendritic cells widely used in immunological studies as a convenient and easy approach after isolation of mononuclear cells directly from peripheral blood mononuclear cells (PBMC). Both the purification and cell culture of monocytes impact on the differentiation of monocytes into moDCs. The methodology to isolate and differentiate monocytes into moDCs is still controversial. We aimed to compare three different protocols for monocyte isolation from PBMC: 1) Cold-aggregation; 2) Percoll gradient; and 3) Magnetic beads cell-enrichment. Additionally we also compared four different monocyte differentiation and culture techniques: 1) Cell culture media; 2) Serum sources; 3) required GM-CSF and IL-4 concentrations; 4) Cell culture systems. We used flow cytometry analysis of light scattering and/or expression of pan surface markers, such as CD3, CD14 and CD209 to determine isolation/differentiation degree. Purified PBMC followed by two steps of cold aggregation, yielded cell viability around 95% with poor monocyte enrichment (monocytes increase vs. lymphocytes reduction was not statistically significant, p>0.05). Conversely, monocyte isolation from PBMC with discontinuous Percoll gradient generated around 50% cell viability. Albeit, we observed a significant reduction (p≤0.05) of lymphocytes contaminants. The magnetic beads cell-enrichment yield cell viability higher than 95%, as high as a significant lymphocyte depletion (p≤0.005) when compared to all other techniques employed. The moDCs showed better differentiation based on increased CD209 expression, but lower CD14 levels, when cells were cultured in RPMI medium plus 500IU/mL of both GM-CSF and IL-4 in a semi-adherent fashion. Serum sources showed no influence on the culture performance. In conclusion, the magnetic beads cell-enrichment showed superior cell viability, indicating that this approach is a better choice to isolate monocytes, and moDCs cultured afterwards in appropriate medium, serum, cytokines and culture system might influence the monocytes differentiation into moDC.


Assuntos
Separação Celular/métodos , Células Dendríticas/citologia , Monócitos/citologia , Antígenos CD/metabolismo , Diferenciação Celular , Sobrevivência Celular , Células Cultivadas , Citometria de Fluxo , Fluorescência , Humanos , Monócitos/metabolismo , Espalhamento de Radiação
16.
PLoS One ; 15(4): e0231223, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32298286

RESUMO

We observed prominent effects of doxorubicin (Dox), an anthracycline widely used in anti-cancer therapy, on the aggregation and intracellular distribution of both partners of the H2A-H2B dimer, with marked differences between the two histones. Histone aggregation, assessed by Laser Scanning Cytometry via the retention of the aggregates in isolated nuclei, was observed in the case of H2A. The dominant effect of the anthracycline on H2B was its massive accumulation in the cytoplasm of the Jurkat leukemia cells concomitant with its disappearance from the nuclei, detected by confocal microscopy and mass spectrometry. A similar effect of the anthracycline was observed in primary human lymphoid cells, and also in monocyte-derived dendritic cells that harbor an unusually high amount of H2B in their cytoplasm even in the absence of Dox treatment. The nucleo-cytoplasmic translocation of H2B was not affected by inhibitors of major biochemical pathways or the nuclear export inhibitor leptomycin B, but it was completely diminished by PYR-41, an inhibitor with pleiotropic effects on protein degradation pathways. Dox and PYR-41 acted synergistically according to isobologram analyses of cytotoxicity. These large-scale effects were detected already at Dox concentrations that may be reached in the typical clinical settings, therefore they can contribute both to the anti-cancer mechanism and to the side-effects of this anthracycline.


Assuntos
Citoplasma/metabolismo , Doxorrubicina/farmacologia , Histonas/metabolismo , Transporte Ativo do Núcleo Celular , Antraciclinas/farmacologia , Antineoplásicos/farmacologia , Núcleo Celular/metabolismo , Proliferação de Células , Ácidos Graxos Insaturados/metabolismo , Humanos , Células Jurkat , Citometria de Varredura a Laser , Espectrometria de Massas , Microscopia Confocal , Monócitos/citologia
17.
J Vis Exp ; (158)2020 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-32310228

RESUMO

Chemotaxis is receptor-mediated guidance of cells along a chemical gradient, whereas chemokinesis is the stimulation of random cell motility by a chemical. Chemokinesis and chemotaxis are fundamental for the mobilization and deployment of immune cells. For example, chemokines (chemotactic cytokines) can rapidly recruit circulating neutrophils and monocytes to extravascular sites of inflammation. Chemoattractant receptors belong to the large family of G protein-coupled receptors. How chemoattractant (i.e., ligand) gradients direct cell migration via G protein-coupled receptor signaling is not yet fully understood. In the field of immunology, neutrophils are popular model cells for studying chemotaxis in vitro. Here we describe a real-time two-dimensional (2D) chemotaxis assay tailored for mouse resident macrophages, which have traditionally been more difficult to study. Macrophages move at a slow pace of ~1 µm/min on a 2D surface and are less well suited for point-source migration assays (e.g., migration towards the tip of a micropipette filled with chemoattractant) than neutrophils or Dictyostelium discoideum, which move an order of magnitude faster. Widely used Transwell assays are useful for studying the chemotactic activity of different substances, but do not provide information on cell morphology, velocity, or chemotactic navigation. Here we describe a time-lapse microscopy-based macrophage chemotaxis assay that allows quantification of cell velocity and chemotactic efficiency and provides a platform to delineate the transducers, signal pathways, and effectors of chemotaxis.


Assuntos
Quimiotaxia , Macrófagos/citologia , Imagem com Lapso de Tempo/métodos , Animais , Dictyostelium/citologia , Macrófagos/metabolismo , Camundongos , Monócitos/citologia , Neutrófilos/citologia , Receptores Acoplados a Proteínas-G/metabolismo , Transdução de Sinais
18.
Rev. andal. med. deporte ; 13(1): 40-44, mar. 2020. ilus, tab, graf
Artigo em Inglês | IBECS | ID: ibc-193416

RESUMO

Phagocytic cells constitute the first defense line against the diversity of infectious agents. The effects of aging on the immune function - immunosenescence - affect the phagocytic capacity of neutrophils and monocytes/macrophages and result in increased risk to cancer and other diseases. The aim of this review was to assess the functional aspects of the innate system cells in aging. Evidence brought about by this review suggests that resistance training is a useful therapy to mitigate the adverse effects of the innate immune system aging process. Resistance training is consistently recommended as assistent strategy for prevention of the inflamaging and associated chronic diseases, but establishing adequate program is still in demand. In addition, future studies are needed to improve our understanding of the resistance training-induced mechanisms underlying changes in phagocytic activity in the elderly


Las células fagocíticas constituyen la primera línea de defensa contra los agentes infecciosos. Los efectos del envejecimiento sobre la función inmune - inmunosenescencia - afectan la capacidad fagocítica de neutrófilos y monocitos/macrófagos y resultan en riesgo aumentado para el cáncer y otras enfermedades. El objetivo de esta revisión fue evaluar los aspectos funcionales de las células del sistema innato en el envejecimiento. Las evidencias revisadas sugieren que el entrenamiento de resistencia es una terapia útil para atenuar los efectos adversos del proceso de envejecimiento del sistema inmune innato. Se recomienda el entrenamiento de resistencia continuamente como estrategia complementaria para la prevención de la inflamación y de las enfermedades crónicas asociadas, pero hay que establecer el programa adecuado. Además, se necesitan más investigaciones para mejorar nuestra comprensión de los mecanismos modulados por el entrenamiento de resistencia que inducen a los cambios en la actividad fagocítica en las personas mayores


As células fagocitárias constituem a primeira linha de defesa contra agentes infecciosos. Os efeitos do envelhecimento sobre a função imune - imunossenescência - afetam a capacidade fagocítica de neutrófilos e monócitos/macrófagos e resultam em aumento do risco para câncer e outras doenças. O objetivo desta revisão foi avaliar os aspectos funcionais das células do sistema inato durante o envelhecimento. Os estudos revisados sugerem que o treinamento resistido é uma terapia útil para atenuar os efeitos adversos do processo de envelhecimento do sistema imune inato. Recomenda-se que o treinamento resistido seja aplicado continuamente como estratégia complementar para a prevenção da inflamação e doenças crônicas associadas, porém deve-se estabelecer o programa adequado. Ressalta-se ainda que, são necessários mais estudos para melhorar a compreensão sobre os mecanismos modulados pelo treinamento resistido que induzem a alterações na atividade fagocítica em idosos


Assuntos
Humanos , Macrófagos/citologia , Treinamento de Resistência , Monócitos/imunologia , Neutrófilos/imunologia , Imunossenescência/fisiologia , Monócitos/citologia , Neutrófilos/citologia , Doença Crônica
19.
Life Sci ; 249: 117518, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32147432

RESUMO

AIMS: The objectives of the present study were to investigate the mechanisms of Ninj-1 regulation in TNFα-activated human endothelial cells (HEC), and to test if Amlodipine (AML) ameliorates the inflammatory stress by decreasing Ninj-1 expression. MAIN METHODS: TNFα-activated HEC with/without AML (0.1 µM and 1 µM) were used. TNFα-receptor 1 (TNFR1) was silenced and inhibitors for oxidative stress (N-acetyl cysteine), endoplasmic reticulum stress (salubrinal, 4-phenyl butyric acid), or NF-kB (Bay 11-7085) and p38 MAPK (SB203580) were used. Levels of Ninj-1, TNFR1, monocyte adhesion, endoplasmic reticulum stress (ERS) sensors, NADPH oxidase- and mitochondria-derived oxidative species were evaluated. KEY FINDINGS: The novel findings that we report here are: (i) silencing the endothelial TNFR1 leads to decreased Ninj-1 expression and diminished monocyte adhesion; (ii) increased oxidative stress, ERS and NF-kB activation enhance Ninj-1 expression and monocyte adhesion; (iii) up-regulation of endothelial Ninj-1 expression stimulates monocytes adhesion to TNFα - activated HEC; (iv) AML diminishes monocyte adhesion by reducing Ninj-1 expression through mechanisms involving the decrease of NADPH oxidase and mitochondria-dependent oxidative stress, ERS and NF-kB. In addition, AML alleviates apoptosis by reducing the pro-apoptotic CHOP expression and re-establishing the mitochondrial transmembrane potential. SIGNIFICANCE: The results of the present study suggest that Ninj-1 and the proteins involved in its regulation can be considered therapeutic targets for the alleviation of inflammation- dependent disorders. In addition, we demonstrate that some of the benefic effects of AML can be achieved through regulation of Ninj-1.


Assuntos
Anlodipino/farmacologia , Moléculas de Adesão Celular Neuronais/fisiologia , Adesão Celular/fisiologia , Monócitos/citologia , Fatores de Crescimento Neural/fisiologia , Receptores Tipo I de Fatores de Necrose Tumoral/fisiologia , Fator de Necrose Tumoral alfa/fisiologia , Regulação para Cima , Vasodilatadores/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Inativação Gênica , Células Endoteliais da Veia Umbilical Humana , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , NADPH Oxidases/metabolismo , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/genética
20.
Proc Natl Acad Sci U S A ; 117(15): 8573-8583, 2020 04 14.
Artigo em Inglês | MEDLINE | ID: mdl-32220961

RESUMO

Dicer is a ribonuclease III enzyme in biosynthesis of micro-RNAs (miRNAs). Here we describe a regulation of Dicer expression in monocytic cells, based on proteolysis. In undifferentiated Mono Mac 6 (MM6) cells, full-length Dicer was undetectable; only an ∼50-kDa fragment appeared in Western blots. However, when MM6 cells were treated with zymosan or LPS during differentiation with TGF-ß and 1,25diOHvitD3, full-length Dicer became abundant together with varying amounts of ∼170- and ∼50-kDa Dicer fragments. Mass spectrometry identified the Dicer fragments and showed cleavage about 450 residues upstream from the C terminus. Also, PGE2 (prostaglandin E2) added to differentiating MM6 cells up-regulated full-length Dicer, through EP2/EP4 and cAMP. The TLR stimuli strongly induced miR-146a-5p, while PGE2 increased miR-99a-5p and miR-125a-5p, both implicated in down-regulation of TNFα. The Ser protease inhibitor AEBSF (4-[2-aminoethyl] benzene sulfonyl fluoride) up-regulated full-length Dicer, both in MM6 cells and in primary human blood monocytes, indicating a specific proteolytic degradation. However, AEBSF alone did not lead to a general increase in miR expression, indicating that additional mechanisms are required to increase miRNA biosynthesis. Finally, differentiation of monocytes to macrophages with M-CSF or GM-CSF strongly up-regulated full-length Dicer. Our results suggest that differentiation regimens, both in the MM6 cell line and of peripheral blood monocytes, inhibit an apparently constitutive Dicer proteolysis, allowing for increased formation of miRNAs.


Assuntos
Diferenciação Celular , RNA Helicases DEAD-box/metabolismo , MicroRNAs/metabolismo , Monócitos/metabolismo , Prostaglandina-E Sintases/metabolismo , Proteólise , Ribonuclease III/metabolismo , Células Cultivadas , RNA Helicases DEAD-box/genética , Dinoprostona/farmacologia , Hematopoese , Humanos , Lipopolissacarídeos/farmacologia , MicroRNAs/genética , Monócitos/citologia , Monócitos/efeitos dos fármacos , Prostaglandina-E Sintases/genética , Receptores de Prostaglandina E Subtipo EP2/metabolismo , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Ribonuclease III/genética , Zimosan/farmacologia
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