RESUMEN
PURPOSE: To examine the impact of polyphenol supplementation on the recruitment, mobilization, and activation of monocyte subsets after resistance exercise. METHODS: Thirty-eight recreationally active males (22.1 ± 3.1 yr; 173.9 ± 7.9 cm; 77.8 ± 14.5 kg) were assigned to 28 d of polyphenol blend (PPB) supplementation, placebo (PL), or control (CON). Blood samples were obtained before (PRE) postresistance exercise, immediately (IP) postresistance exercise, 1 h (1H) postresistance exercise, 5 h (5H) postresistance exercise, 24 h (24H) postresistance exercise, and 48 h (48H) postresistance exercise (PPB/PL) or rest (CON). Fine-needle biopsies were obtained from the vastus lateralis at PRE, 1H, 5H, and 48H. Circulating concentrations of macrophage chemoattractant protein-1 (MCP-1) and fractalkine, as well as intramuscular MCP-1 were analyzed via multiplex assay. Changes in the proportions and expression of CD11b on monocyte subsets were assessed via flow cytometry. RESULTS: Circulating MCP-1 increased in PPB and PL at IP with further increases at 5H. Intramuscular MCP-1 was increased at 1H, 5H, and 48H in all groups. Classical monocyte proportions were reduced in PPB and PL at IP, and increased at 1H. Nonclassical monocytes were increased in PPB and PL at IP, whereas intermediate monocytes were increased at IP, and reduced at 1H. Intermediate monocytes were increased in PPB at 24H and 48H. CD11b expression was reduced on PPB compared with PL and CON at PRE on intermediate and nonclassical monocytes. CONCLUSIONS: Resistance exercise may elicit selective mobilization of intermediate monocytes at 24H and 48H, which may be mediated by tissue damage. Additionally, polyphenol supplementation may suppress CD11b expression on monocyte subsets at rest.
Asunto(s)
Antioxidantes/administración & dosificación , Suplementos Dietéticos , Monocitos/metabolismo , Polifenoles/administración & dosificación , Músculo Cuádriceps/metabolismo , Entrenamiento de Fuerza , Antígeno CD11b/sangre , Quimiocina CCL2/sangre , Quimiocina CCL2/metabolismo , Quimiocina CX3CL1/sangre , Humanos , Antígeno de Macrófago-1/sangre , Masculino , Factores de Tiempo , Adulto JovenRESUMEN
OBJECTIVE: The objective was to investigate the effect of commonly used inhaled corticosteroids on white blood cell count (WBC) and to examine the mechanisms involved. METHODS: This randomized comparative study comprised 60 healthy adults. We measured the effects of budesonide (by face mask inhalation or aerosol inhaler), fluticasone (by inhaler), and saline inhalation (control) on WBC and the differential leukocyte count, especially the absolute neutrophil count (ANC). To elucidate the mechanisms involved, we measured the expression of the adhesion neutrophil ligands Mac-1 (CD11b) and L-selectin (CD62L), and granulocyte colony-stimulating factor serum levels. RESULTS: Six hours after a single-dose inhalation of budesonide, mean increases of 23.4% in WBC (95% confidence interval [CI], 11.3-35.4) and 30.1% in ANC (95% CI, 7.2-53.0) were noted. The percentage of neutrophils increased from 54.6% to 58.1% (P< .001). Inhaled fluticasone increased WBC and ANC by 12.6% (95% CI, 1.5-23.7) and 22.7% (95% CI, 6.2-39.2), respectively (P< .01 for both). The absolute lymphocyte and eosinophil counts did not change significantly from baseline. The expression of Mac-1 and L-selectin decreased by 51.0% (P< .01) and 30.9% (P= .02), respectively, following face mask inhalation of budesonide and by 39.8% (P= .01) and 17.4% (P= .17), respectively, following inhalation of fluticasone. No significant changes in granulocyte colony-stimulating factor levels were noted. CONCLUSIONS: Glucocorticoid inhalation increases WBC by increasing ANC. Reduced neutrophil adhesion to the endothelial surface, mediated by decreased adhesion molecule expression on neutrophils, is a plausible mechanism. Physicians should be aware of the effect of inhaled corticosteroids on WBC, as it may influence clinical decisions, especially in the emergency department.
Asunto(s)
Corticoesteroides/farmacología , Budesonida/farmacología , Fluticasona/farmacología , Selectina L/sangre , Recuento de Leucocitos , Antígeno de Macrófago-1/sangre , Administración por Inhalación , Corticoesteroides/administración & dosificación , Adulto , Budesonida/administración & dosificación , Femenino , Fluticasona/administración & dosificación , Factor Estimulante de Colonias de Granulocitos/sangre , Humanos , Masculino , Persona de Mediana Edad , Neutrófilos/efectos de los fármacos , Adulto JovenRESUMEN
UNLABELLED: The innate immune response is generally considered to have an important role in tissue remodeling after resistance exercise. PURPOSE: The purpose of this study was to compare changes in markers of monocyte recruitment after an acute bout of high-intensity (HVY) versus high-volume (VOL) lower-body resistance exercise. METHODS: Ten resistance-trained men (24.7 ± 3.4 yr, 90.1 ± 11.3 kg, 176.0 ± 4.9 cm) performed each protocol in a randomized, counterbalanced order. Blood samples were collected at baseline, immediately (IP), 30 min (30P), 1 h (1H), 2 h (2H), and 5 h (5H) postexercise. Plasma concentrations of monocyte chemoattractant protein 1 (MCP-1), tumor necrosis factor alpha (TNF-α), myoglobin, and cortisol were measured via assay. Tumor necrosis factor receptor 1 (TNFr1), macrophage-1 antigen (cluster of differentiation 11b [CD11b]), and C-C chemokine receptor 2 (CCR2) expression levels were measured using flow cytometry. TNFr1 and CD11b were assessed on CD14CD16 monocytes, whereas CCR2 was assessed on CD14 monocytes. RESULTS: Plasma myoglobin concentrations were significantly greater after HVY compared with VOL (P < 0.001). Changes in plasma TNF-α, MCP-1, and expression levels of CCR2 and CD11b were similar between HVY and VOL. When collapsed across groups, TNF-α was significantly increased at IP, 30P, 1H, and 2H (P values < 0.05), whereas MCP-1 was significantly elevated at all postexercise time points (P values < 0.05). CCR2 expression on CD14 monocytes was significantly lower at IP, 1H, 2H, and 5H (P values < 0.05). CD11b expression on CD14 CD16 was significantly greater at IP (P < 0.014) and 1H (P = 0.009). TNFr1 expression did not differ from baseline at any time point. Plasma cortisol concentrations did not seem to be related to receptor expression. CONCLUSIONS: Results indicate that both HVY and VOL protocols stimulate a robust proinflammatory response. However, no differences were noted between resistance exercise training paradigms.
Asunto(s)
Monocitos/metabolismo , Entrenamiento de Fuerza/métodos , Antígeno CD11b/sangre , Quimiocina CCL2/sangre , Humanos , Hidrocortisona/sangre , Inmunidad Innata/fisiología , Antígeno de Macrófago-1/sangre , Masculino , Mioglobina/sangre , Receptores CCR2/sangre , Receptores Tipo I de Factores de Necrosis Tumoral/sangre , Factor de Necrosis Tumoral alfa/sangre , Adulto JovenRESUMEN
The clinical management of paroxysmal nocturnal hemoglobinuria (PNH), a rare but life-threatening hematologic disease, has fundamentally improved with the introduction of a therapeutic that prevents complement-mediated intravascular hemolysis. However, a considerable fraction of PNH patients show insufficient treatment response and remain transfusion dependent. Because the current treatment only prevents C5-induced lysis but not upstream C3 activation, it has been speculated that ongoing opsonization with C3 fragments leads to recognition and phagocytosis of PNH erythrocytes by immune cells. Here, for the first time, we provide experimental evidence for such extravascular hemolysis and demonstrate that PNH erythrocytes from anti-C5-treated patients are phagocytosed by activated monocytes in vitro. Importantly, we show that this uptake can be mediated by the end-stage opsonin C3dg, which is not traditionally considered a phagocytic marker, via interaction with complement receptor 3 (CR3). Interaction studies confirmed that C3dg itself can act as a ligand for the binding domain of CR3. The degree of C3dg-mediated erythrophagocytosis in samples from different PNH patients correlated well with the individual level of C3dg opsonization. This finding may guide future treatment options for PNH but also has potential implications for the description and management of other complement-mediated diseases.
Asunto(s)
Hemoglobinuria Paroxística/sangre , Hemoglobinuria Paroxística/inmunología , Fragmentos de Péptidos/sangre , Complemento C3b , Eritrocitos/inmunología , Eritrocitos/patología , Hemólisis/inmunología , Humanos , Antígeno de Macrófago-1/sangre , Modelos Inmunológicos , Proteínas Opsoninas/sangre , Fagocitosis/inmunologíaRESUMEN
Intravenous immunoglobulin (IVIG) decreases neutrophil adhesion to endothelium and red blood cell-neutrophil interactions in sickle cell mice undergoing vaso-occlusion. In this Phase I clinical trial of sickle cell anemia (SCA) patients admitted with pain crisis, we evaluated the status of adhesion molecules on neutrophils in control and IVIG-treated subjects pre- and post-infusion up to 800 mg/kg, the same dose used in murine studies. Mac-1 function significantly decreased from baseline in the low-dose IVIG (200-400 mg/kg) cohorts. IVIG-related adverse events may have occurred in the high-dose (600-800 mg/kg) cohorts. There were no significant increases in neutrophil and leukocyte counts, suggesting that IVIG may more selectively inhibit Mac-1 function as opposed to neutrophil adhesion. This study provides the first in-human validation of pre-clinical murine studies that IVIG can decrease Mac-1 function.
Asunto(s)
Síndrome Torácico Agudo/tratamiento farmacológico , Anemia de Células Falciformes/tratamiento farmacológico , Inmunoglobulinas Intravenosas/uso terapéutico , Antígeno de Macrófago-1/sangre , Neutrófilos/efectos de los fármacos , Dolor/tratamiento farmacológico , Síndrome Torácico Agudo/sangre , Síndrome Torácico Agudo/complicaciones , Síndrome Torácico Agudo/fisiopatología , Adolescente , Adulto , Anemia de Células Falciformes/sangre , Anemia de Células Falciformes/complicaciones , Anemia de Células Falciformes/fisiopatología , Adhesión Celular/efectos de los fármacos , Niño , Esquema de Medicación , Femenino , Humanos , Masculino , Activación Neutrófila/efectos de los fármacos , Neutrófilos/metabolismo , Neutrófilos/patología , Dolor/sangre , Dolor/complicaciones , Dolor/fisiopatologíaRESUMEN
Neutrophil recruitment is known to be a rate-limiting step in mediating tissue injury in severe acute pancreatitis (AP). However, the signalling mechanisms controlling inflammation and organ damage in AP remain elusive. Herein, we examined the role of Ras signalling in AP. Male C57BL/6 mice were treated with a Ras inhibitor (farnesylthiosalicylic acid, FTS) before infusion of taurocholate into the pancreatic duct. Pancreatic and lung tissues as well as blood were collected 24 h after pancreatitis induction. Pretreatment with FTS decreased serum amylase levels by 82% and significantly attenuated acinar cell necrosis, tissue haemorrhage and oedema formation in taurocholate-induced pancreatitis. Inhibition of Ras signalling reduced myeloperoxidase (MPO) levels in the inflamed pancreas by 42%. In addition, administration of FTS decreased pancreatic levels of CXC chemokines as well as circulating levels of interleukin-6 and high-mobility group box 1 in animals exposed to taurocholate. Moreover, treatment with FTS reduced taurocholate-induced MPO levels in the lung. Inhibition of Ras signalling had no effect on neutrophil expression of Mac-1 in mice with pancreatitis. Moreover, FTS had no direct impact on trypsin activation in isolated pancreatic acinar cells. These results indicate that Ras signalling controls CXC chemokine formation, neutrophil recruitment and tissue injury in severe AP. Thus, our findings highlight a new signalling mechanism regulating neutrophil recruitment in the pancreas and suggest that inhibition of Ras signalling might be a useful strategy to attenuate local and systemic inflammation in severe AP.
Asunto(s)
Inhibidores Enzimáticos/uso terapéutico , Farnesol/análogos & derivados , Infiltración Neutrófila/efectos de los fármacos , Páncreas/efectos de los fármacos , Pancreatitis Aguda Necrotizante/prevención & control , Proteínas Proto-Oncogénicas p21(ras)/antagonistas & inhibidores , Salicilatos/uso terapéutico , Transducción de Señal/efectos de los fármacos , Células Acinares/efectos de los fármacos , Células Acinares/inmunología , Células Acinares/metabolismo , Células Acinares/patología , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Quimiocinas/antagonistas & inhibidores , Quimiocinas/sangre , Quimiocinas/metabolismo , Farnesol/uso terapéutico , Proteína HMGB1/sangre , Interleucina-6/sangre , Antígeno de Macrófago-1/sangre , Antígeno de Macrófago-1/metabolismo , Masculino , Ratones Endogámicos C57BL , Terapia Molecular Dirigida , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/metabolismo , Neutrófilos/patología , Páncreas/inmunología , Páncreas/metabolismo , Páncreas/patología , Pancreatitis Aguda Necrotizante/inmunología , Pancreatitis Aguda Necrotizante/metabolismo , Pancreatitis Aguda Necrotizante/patología , Proteínas Proto-Oncogénicas p21(ras)/metabolismoRESUMEN
BACKGROUND: Insufficient leukocyte recruitment may be one reason for the high incidence of life-threatening infections in preterm infants. Since the receptor of advanced glycation end products (RAGE) is a known leukocyte adhesion molecule and highly expressed during early development, we asked whether RAGE plays a role for leukocyte recruitment in preterm and term infants. METHODS: Leukocyte adhesion was analyzed in dynamic flow chamber experiments using isolated leukocytes of cord blood from extremely premature (<30 weeks of gestation), moderately premature (30-35 weeks of gestation) and mature neonates (>35 weeks of gestation) and compared to the results of adults. For fluorescent microscopy leukocytes were labeled with rhodamine 6G. In the respective age groups we also measured the plasma concentration of soluble RAGE (sRAGE) by ELISA and Mac-1 and LFA-1 expression on neutrophils by flow cytometry. RESULTS: The adhesive functions of fetal leukocytes significantly increase with gestational age. In all age groups, leukocyte adhesion was crucially dependent on RAGE. In particular, RAGE was equally effective to mediate leukocyte adhesion when compared to ICAM-1. The plasma levels of sRAGE were high in extremely premature infants and decreased with increasing gestational age. In contrast, expression of ß2-Integrins Mac-1 and LFA-1 which are known ligands for RAGE and ICAM-1 did not change during fetal development. CONCLUSION: We conclude that RAGE is a crucial leukocyte adhesion molecule in both preterm and term infants.
Asunto(s)
Recien Nacido Extremadamente Prematuro/sangre , Leucocitos/metabolismo , Receptores Inmunológicos/sangre , Adulto , Adhesión Celular/inmunología , Femenino , Regulación de la Expresión Génica/inmunología , Humanos , Recien Nacido Extremadamente Prematuro/inmunología , Recién Nacido , Molécula 1 de Adhesión Intercelular/sangre , Molécula 1 de Adhesión Intercelular/inmunología , Leucocitos/inmunología , Leucocitos/patología , Antígeno-1 Asociado a Función de Linfocito/sangre , Antígeno-1 Asociado a Función de Linfocito/inmunología , Antígeno de Macrófago-1/sangre , Antígeno de Macrófago-1/inmunología , Masculino , Receptor para Productos Finales de Glicación Avanzada , Receptores Inmunológicos/inmunologíaRESUMEN
Mediators of monocyte migration, complement receptor-3 (CR3), and chemokine ligand-4 (CCL4) were measured in response to recovery modalities following resistance exercise. Thirty resistance-trained men (23.1 ± 2.9 y; 175.2 ± 7.1 cm; 82.1 ± 8.4 kg) were given neuromuscular electric stimulation (NMES), cold water immersion (CWI), or control (CON) treatments immediately following resistance exercise. Blood samples were obtained preexercise (PRE), immediately (IP), 30 minutes (30 P), 24 hours (24 H), and 48 hours (48 H) after exercise for measurement of circulating CCL4 and CR3 expression on CD14+ monocytes, by assay and flow cytometry. Circulating CCL4 showed no consistent changes. Inferential analysis indicated that CR3 expression was likely greater in CON at 30 P than NMES (90.0%) or CWI (86.8%). NMES was likely lower than CON at 24 H (92.9%) and very likely lower at 48 H (98.7%). Expression of CR3 following CWI was very likely greater than CON (96.5%) at 24 H. The proportion of CR3+ monocytes was likely greater following CWI than NMES (85.8%) or CON (85.2%) at 24 H. The change in proportion of CR3+ monocytes was likely (86.4%) greater following NMES than CON from IP to 30 P. The increased expression of CR3 and increased proportion of CR3+ monocytes following CWI at 24 H indicate a potentially improved ability for monocyte adhesion to the endothelium, possibly improving phagocytosis of damaged tissues.
Asunto(s)
Ejercicio Físico/fisiología , Monocitos/citología , Monocitos/metabolismo , Músculo Esquelético/metabolismo , Músculo Esquelético/fisiología , Adulto , Quimiocina CCL4/sangre , Citometría de Flujo , Humanos , Antígeno de Macrófago-1/sangre , Masculino , Adulto JovenRESUMEN
OBJECTIVE: The platelet glycoprotein Ib-IX (GP Ib-IX) receptor is a well-characterized adhesion receptor supporting hemostasis and thrombosis via interactions with von Willebrand factor. We examine the GP Ib-IX/von Willebrand factor axis in murine polymicrobial sepsis, as modeled by cecal ligation and puncture (CLP). APPROACH AND RESULTS: Genetic absence of the GP Ib-IX ligand, von Willebrand factor, prolongs survival after CLP, but absence of the receptor, GP Ib-IX, does not. Because absence of either von Willebrand factor or GP Ib-IX significantly impairs hemostasis and thrombosis, we sought to define additional GP Ib-IX-dependent pathways impacting survival in the CLP model. We document that the absence of GP Ib-IX leads to reduced platelet-neutrophil and platelet-monocyte interactions. Twenty-four hours after CLP, absence of GP Ib-IX coincides with an alteration in cytokine levels, such as tumor necrosis factor-α secreted by monocytes, and increased macrophage-1 antigen expression by neutrophils. CONCLUSIONS: In contrast to the well-characterized proinflammatory properties of platelets, we describe in the CLP model an anti-inflammatory property associated with platelet GP Ib-IX. Thus, a single platelet receptor displays a dual modulatory role in both the thrombotic and inflammatory pathways associated with polymicrobial sepsis. In sharing leucine-rich motifs with toll-like receptors, platelet GP Ib-IX can be considered a multifunctional participant in hemostasis, thrombosis, and the inflammatory cascade. The results highlight a dynamic role for platelets in systemic inflammation and add to the complex pathophysiologic events that occur during the dysregulated coagulation and inflammation associated with sepsis.
Asunto(s)
Plaquetas/metabolismo , Complejo GPIb-IX de Glicoproteína Plaquetaria/metabolismo , Sepsis/metabolismo , Animales , Plaquetas/inmunología , Ciego/microbiología , Ciego/cirugía , Comunicación Celular , Modelos Animales de Enfermedad , Hemostasis , Humanos , Interleucina-4/genética , Interleucina-4/metabolismo , Ligandos , Ligadura , Antígeno de Macrófago-1/sangre , Masculino , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Monocitos/metabolismo , Neutrófilos/metabolismo , Complejo GPIb-IX de Glicoproteína Plaquetaria/genética , Sepsis/sangre , Sepsis/genética , Sepsis/inmunología , Sepsis/microbiología , Transducción de Señal , Trombosis/sangre , Trombosis/metabolismo , Factores de Tiempo , Factor de Necrosis Tumoral alfa/sangre , Factor de von Willebrand/genética , Factor de von Willebrand/metabolismoRESUMEN
The inflammatory response to muscle-damaging exercise requires monocyte mobilization and adhesion. Complement receptor type 3 (CR3) and macrophage inflammatory protein (MIP)-1ß enables monocyte recruitment, adhesion, and subsequent infiltration into damaged muscle tissue. The purpose of this study was to examine the effects of cold water immersion (CWI) and/or ß-hydroxy-ß-methylbutyrate free acid (HMB-FA) on CR3 expression and MIP-1ß concentration after four sets of up to 10 repetitions of squat, dead lift, and split squat exercises at 70-80% 1-repetition maximum. Thirty-nine resistance-trained men (22.2 ± 2.5 yr) were randomly divided into four groups: 1) placebo (PL), 2) HMB-FA, 3) HMB-FA-CWI, and 4) PL-CWI. The HMB-FA groups ingested 3 g/day, and CWI groups were submersed into 10-12°C water for 10 min after exercise. Blood was sampled at baseline (PRE), immediately post- (IP), 30 min post- (30P), 24 h post- (24P), and 48 h post (48P)-exercise. Circulating MIP-1ß was assayed and CR3 expression on CD14+ monocytes was measured by flow cytometry. Without treatment, CR3 expression significantly elevated at 30P compared with other time points (P = 0.030-0.047). HMB-FA significantly elevated the percentage of monocytes expressing CR3 between IP and 24P (P = 0.046) and between IP and 48P (P = 0.046). No time effect was observed for MIP-1ß concentration. The recovery modalities showed to attenuate the rise in CR3 following exercise. Additionally, supplementation with HMB-FA significantly elevated the percentage of monocytes expressing CR3 during recovery. Although the time course that inflammatory responses are most beneficial remains to be determined, recovery modalities may alter immune cell mobilization and adhesion mechanisms during tissue recovery.
Asunto(s)
Quimiocina CCL4/sangre , Frío , Inmersión , Antígeno de Macrófago-1/efectos de los fármacos , Monocitos/efectos de los fármacos , Contracción Muscular , Músculo Esquelético/efectos de los fármacos , Entrenamiento de Fuerza , Valeratos/farmacología , Agua , Adulto , Método Doble Ciego , Florida , Humanos , Antígeno de Macrófago-1/sangre , Masculino , Monocitos/inmunología , Monocitos/metabolismo , Músculo Esquelético/inmunología , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Recuperación de la Función , Factores de Tiempo , Adulto JovenRESUMEN
Individual ventricular assist device (VAD) design may affect leukocytes and impact immunity. Few studies have presented leukocyte and infection profiles in VAD patients over the course of the implant period. CD11b (MAC-1) expression on granulocytes is an indicator of activation during inflammation, mediating extravasation and the release of reactive oxygen species in tissue. No reported studies have presented MAC-1 expression on circulating granulocytes in VAD patients. Fifty-six patients implanted at a single center with a HeartMate II (HMII; n = 32), HeartWare (HW; n = 12), or Thoratec pneumatic VAD (PVAD; n = 12) between 1999 and 2011 were followed for 120 days of support. The leukocyte profiles and infectious events of all patients were evaluated; additionally, a subset had MAC-1 expression on circulating granulocytes was measured (HMIIâ n = 9; HWâ n = 7; PVADâ n = 4). All groups exhibited a significant peak in leukocyte numbers at postoperative day (POD) 14 while simultaneously experiencing a significant decrease in hematocrit. HMII patients exhibited a 3.2-fold increase in granulocyte MAC-1 expression at POD 14, and the temporal trend over the implant period differed from that experienced by HW patients. Further, HW patients experienced significantly fewer infection events. Alterations in leukocyte profiles and granulocyte activation experienced by VAD patients appear to be device-specific. Elevations in leukocyte activation may be related to an increased risk for infection, although the specific relationship between these phenomena in this patient group is not known.
Asunto(s)
Granulocitos/inmunología , Insuficiencia Cardíaca/terapia , Corazón Auxiliar , Leucocitos/inmunología , Implantación de Prótesis/instrumentación , Función Ventricular Izquierda , Adulto , Anciano , Biomarcadores/sangre , Femenino , Granulocitos/metabolismo , Insuficiencia Cardíaca/diagnóstico , Insuficiencia Cardíaca/fisiopatología , Corazón Auxiliar/efectos adversos , Hematócrito , Humanos , Recuento de Leucocitos , Leucocitos/metabolismo , Antígeno de Macrófago-1/sangre , Masculino , Persona de Mediana Edad , Pennsylvania , Valor Predictivo de las Pruebas , Diseño de Prótesis , Implantación de Prótesis/efectos adversos , Infecciones Relacionadas con Prótesis/sangre , Infecciones Relacionadas con Prótesis/diagnóstico , Infecciones Relacionadas con Prótesis/inmunología , Factores de Riesgo , Infección de la Herida Quirúrgica/sangre , Infección de la Herida Quirúrgica/diagnóstico , Infección de la Herida Quirúrgica/inmunología , Factores de Tiempo , Resultado del TratamientoRESUMEN
Circulating platelet-leukocyte hetero-aggregates play an important role in acute cardiovascular events and hypersensitivity reactions. The association involves the receptor families of selectins and integrin. The objective of this study was to investigate the role of CD11b/CD18 integrin (Mac-1) in hetero-aggregate formation and search for a counter-receptor on platelets ready to interact with Mac-1. As a model of leukocytes, Mac-1 presenting Chinese hamster ovary (CHO) cells were used to evaluate the role of Mac-1 in hetero-aggregate formation. The amount of CHO cell-bound active and inactive platelets was measured by flow cytometry, while the counter-receptors on platelets were identified via using blocking antibodies. We observed significant platelet adhesion on Mac-1-bearing cells when platelet-rich plasma or activated platelets were present. Inactive platelets did not adhere to Mac-1-bearing cells. Addition of fibrinogen, a ligand of Mac-1 significantly increased platelet binding. CD40L was demonstrated to act similarly on Mac-1. Inhibition of platelet GpIIb/IIIa completely abolished CHO cell-platelet aggregation. In our study, we have shown for the first time that Mac-1 mediates the formation of hetero-aggregates without selectin tethering when Mac-1 ligands such as fibrinogen or CD40L are present and blockers of platelet GpIIb/IIIa are able to diminish this interaction.
Asunto(s)
Plaquetas/metabolismo , Leucocitos/metabolismo , Antígeno de Macrófago-1/sangre , Agregación Plaquetaria/fisiología , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/antagonistas & inhibidores , Animales , Células CHO , Cricetinae , Humanos , Antígeno de Macrófago-1/genética , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , TransfecciónRESUMEN
Prasugrel, through its active metabolite, reduces atherothrombosis and its clinical manifestations by inhibiting platelet activation and aggregation. Platelets also contribute to inflammation through interaction with different classes of leukocytes. We investigated whether the inhibitory effect of prasugrel on platelets also counteract inflammatory responses. The effect of prasugrel active metabolite, R-138727, was investigated on platelet P-selectin expression, platelet adhesion to polymorphonuclear leukocytes (PMN) and monocytes (MN) and Mac-1 expression in PMN and MN, in vitro, in human cells. The ex vivo effect of prasugrel administration on P-selectin, thromboxane (TXB)2 formation, platelet-PMN conjugates and Mac-1 expression in PMN triggered by PAR-4 agonist peptide was examined in whole blood from healthy mice as well as from mice in which an acute inflammatory reaction was induced by treatment with endotoxin. The effect of prasugrel on inflammatory markers in endotoxin-treated animals was also tested in vivo. R-138727 inhibited agonist-stimulated expression of platelet P-selectin, platelet-PMN and platelet-MN adhesion and platelet-dependent Mac-1 expression in leukocytes. Addition of aspirin did not modify the inhibitory effect elicited by R-138727. Treatment of mice with prasugrel resulted in a profound inhibition of platelet P-selectin expression, TXB2 production, platelet-PMN adhesion and Mac-1 expression in PMN induced by ex vivo stimulation with PAR-4 agonist peptide of whole blood from healthy or endotoxin-treated mice. Measurement of markers revealed that prasugrel reduced TXB2 and tumour necrosis factor-α synthesis and increased nitric oxide metabolites in endotoxin-treated mice in vivo. In conclusion, prasugrel reduces platelet interactions with PMN and MN. Through these effects prasugrel may curb platelet-mediated inflammatory responses.
Asunto(s)
Antiinflamatorios/farmacología , Plaquetas/efectos de los fármacos , Mediadores de Inflamación/sangre , Leucocitos/efectos de los fármacos , Piperazinas/farmacología , Inhibidores de Agregación Plaquetaria/farmacología , Antagonistas del Receptor Purinérgico P2Y/farmacología , Choque Séptico/tratamiento farmacológico , Tiofenos/farmacología , Animales , Aspirina/farmacología , Biomarcadores/sangre , Plaquetas/inmunología , Modelos Animales de Enfermedad , Regulación hacia Abajo , Humanos , Leucocitos/inmunología , Antígeno de Macrófago-1/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Óxido Nítrico/sangre , Adhesividad Plaquetaria/efectos de los fármacos , Pruebas de Función Plaquetaria , Clorhidrato de Prasugrel , Selenoproteína P/sangre , Choque Séptico/sangre , Choque Séptico/inmunología , Tromboxano B2/sangre , Factor de Necrosis Tumoral alfa/sangreRESUMEN
BACKGROUND: Chronic periodontitis (CP) and aggressive periodontitis (AP) are inflammatory diseases and the main cause of dental loss in adults. We aimed to investigate the expression of adhesion molecules and the source of proinflammatory and anti-inflammatory cytokines in circulating mononuclear cells from patients with CP and AP. METHODS: Peripheral blood mononuclear cells from healthy controls and CP or AP patients were collected. The expression of the cell adhesion molecules CD11a and CD11b, and the cellular sources of interleukin (IL)-4, IL-10, IL-12, interferon-γ, and tumor necrosis factor-α by distinct subpopulations of circulating leukocytes were determined using flow cytometry. RESULTS: The expression of CD11a, but not CD11b, was significantly higher within the CD4(+) and CD8(+) T cells in CP and AP than in healthy controls. The frequencies of tumor necrosis factor-α-expressing CD4(+) T cells and CD14(+) cells were higher in AP and CP, compared to healthy controls, respectively. Moreover, the frequency of IL-10 expressing CD14(+) cells was higher in CP, but not AP, compared to healthy controls CD4(+) T cells committed to IL-4 production was higher in CP than in healthy controls. CONCLUSION: These results suggest the participation of CD11a in the pathogenesis of periodontal lesions and show distinct cellular sources of immunoregulatory cytokines in AP versus CP.
Asunto(s)
Periodontitis Agresiva/sangre , Periodontitis Crónica/sangre , Citocinas/sangre , Leucocitos Mononucleares/inmunología , Adolescente , Adulto , Periodontitis Agresiva/inmunología , Antígenos CD/sangre , Antígenos CD19/sangre , Antígenos de Diferenciación de Linfocitos T/sangre , Antígeno CD11a/sangre , Antígeno CD11b/sangre , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Periodontitis Crónica/inmunología , Femenino , Humanos , Mediadores de Inflamación/inmunología , Molécula 1 de Adhesión Intercelular/sangre , Interferón gamma/sangre , Interleucina-10/sangre , Interleucina-12/sangre , Interleucina-4/sangre , Lectinas Tipo C/sangre , Leucocitos/clasificación , Receptores de Lipopolisacáridos/sangre , Antígeno-1 Asociado a Función de Linfocito/sangre , Antígeno de Macrófago-1/sangre , Masculino , Persona de Mediana Edad , Monocitos/inmunología , Factor de Necrosis Tumoral alfa/análisis , Adulto JovenAsunto(s)
Integrina alfaXbeta2/sangre , Antígeno de Macrófago-1/sangre , Receptores de Complemento 3b/sangre , Receptores de Complemento 3d/sangre , Biomarcadores/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Citometría de Flujo/métodos , Humanos , Integrina alfaXbeta2/química , Integrina alfaXbeta2/fisiología , Síndrome de Deficiencia de Adhesión del Leucocito/diagnóstico , Lupus Eritematoso Sistémico/diagnóstico , Antígeno de Macrófago-1/química , Antígeno de Macrófago-1/fisiología , Fagocitosis , Receptores de Complemento 3b/química , Receptores de Complemento 3b/fisiología , Receptores de Complemento 3d/química , Receptores de Complemento 3d/fisiología , Valores de Referencia , Manejo de EspecímenesRESUMEN
BACKGROUND: Sulfatides are known to be a native ligand of P-selectin. Platelet-leukocyte interaction via the cross-talk between P-selectin and Mac-1 (CD11b/CD18) plays an important role in the mechanism of neointimal thickening after vascular injury such as that seen in post-stent restenosis. However, the roles of sulfatides on restenosis have not been elucidated. METHODS: Serum sulfatide levels, P-selectin expression on the surface of platelets, and activated Mac-1 on the surface of neutrophils were serially measured using both coronary sinus and peripheral blood samples in 21 patients who underwent coronary stent implantation. RESULTS: The trans-cardiac gradient (coronary sinus minus peripheral blood) of the sulfatide levels significantly increased at 15 min (-1.47+/-2.87 to 0.59+/-1.44 nmol/ml, p<0.001), compared to baseline levels. The maximum response of the trans-cardiac gradient of P-selectin expression on the surface of platelets at 15 min after stent implantation (R=0.55, p<0.01), and that of activated Mac-1 on the surface of neutrophils at 48 h (R=0.59, p<0.01), were both positively correlated with that of serum sulfatide levels at 15 min. The angiographic late lumen loss was correlated with the trans-cardiac gradient of sulfatide levels at 15 min (R=0.48, p<0.05), platelet P-selectin expression at 15 min (R=0.42, p<0.05) and activated neutrophil Mac-1 expression at 48 h (R=0.46, p<0.05), but not with values at other sampling points. CONCLUSIONS: Sulfatides may play a physiological role on inflammation in vascular injury and the development of neointimal thickening.
Asunto(s)
Selectina-P/sangre , Sulfoglicoesfingolípidos/sangre , Anciano , Angioplastia Coronaria con Balón , Femenino , Humanos , Inflamación/sangre , Antígeno de Macrófago-1/sangre , Masculino , Persona de Mediana Edad , Selectina-P/metabolismo , Stents , Sulfoglicoesfingolípidos/metabolismo , Túnica Íntima/patologíaRESUMEN
Phagocytosis and oxidative burst are critical host defense mechanisms in which neutrophils clear invading pathogens. Clearing phagocytic neutrophils by triggering apoptosis is an essential process for controlling inflammation. This study elucidates how various exercise bouts with/without hypoxia affected neutrophil bactericidal activity and subsequent apoptosis in humans. Fifteen sedentary males performed six distinct experimental tests in an air-conditioned normobaric hypoxia chamber: two normoxic exercises [strenuous exercise (SE; up to maximal O2 consumption) and moderate exercise (ME; 50% maximal O2 consumption for 30 min) while exposed to 21% O2], two hypoxic exercises (ME for 30 min while exposed to 12% and 15% O2), and two hypoxic exposures (resting for 30 min while exposed to 12% and 15% O2). The results showed that 1) plasma complement-C3a desArg/C4a desArg/C5a concentrations were increased, 2) expressions of L-selectin/lymphocyte functin-associated antigen-1/Mac-1/C5aR on neutrophils were enhanced, 3) phagocytosis of neutrophils to Esherichia coli and release of neutrophil oxidant products by E. coli were elevated, and 4) E. coli-induced phosphotidylserine exposure or caspase-3 activation of neutrophils were promoted immediately and 2 h after both 12% O2 exposure at rest and with ME as well as normoxic SE. Although neither normoxic ME nor breathing 15% O2 at rest influenced these complement- and neutrophil-related immune responses, ME at both 12% and 15% O2 resulted in enhanced complement activation in the blood, expressions of opsonic/complement receptors on neutrophils, or the bactericidal activity and apoptosis of neutrophils. Moreover, the increased neutrophil oxidant production and apoptosis by normoxic SE and hypoxic ME were ameliorated by treating neutrophils with diphenylene iodonium (a NADPH oxidase inhibitor). Therefore, we conclude that ME at 12-15% O2 enhances bactericidal capacity and facilitates the subsequent apoptosis of neutrophils.
Asunto(s)
Apoptosis , Actividad Bactericida de la Sangre , Escherichia coli/patogenicidad , Ejercicio Físico , Hipoxia/microbiología , Neutrófilos/microbiología , Fagocitosis , Estallido Respiratorio , Adulto , Apoptosis/efectos de los fármacos , Biomarcadores/sangre , Actividad Bactericida de la Sangre/efectos de los fármacos , Caspasa 3/metabolismo , Complemento C3a/metabolismo , Complemento C4a/metabolismo , Complemento C5a des-Arginina/metabolismo , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Humanos , Hipoxia/inmunología , Hipoxia/patología , Hipoxia/fisiopatología , Selectina L/sangre , Antígeno-1 Asociado a Función de Linfocito/sangre , Antígeno de Macrófago-1/sangre , Masculino , NADPH Oxidasas/antagonistas & inhibidores , NADPH Oxidasas/metabolismo , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/patología , Compuestos Onio/farmacología , Consumo de Oxígeno , Fagocitosis/efectos de los fármacos , Fosfatidilserinas/metabolismo , Receptor de Anafilatoxina C5a , Receptores de Complemento/sangre , Estallido Respiratorio/efectos de los fármacos , Factores de Tiempo , Adulto JovenRESUMEN
OBJECTIVES: Over one million ragpickers collect and sale recyclable materials from municipal solid wastes (MSW) in India for a living. Since MSW contains a host of pathogenic microorganisms, we investigated the occurrence of airway inflammation and its underlying mechanism in 52 non-smoking female ragpickers (median age 29 yr) and 42 control women matched for age, smoking habit and socioeconomic conditions in Kolkata, eastern India. METHODS: Spontaneously expectorated sputum were stained using the Papanicolau method for cytology, and flow cytometry was used for measurements of surface expression of beta(2) Mac-1 integrin (CD11b/CD18) on leukocytes and P-selectin on platelets. The concentrations of pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) and chemokine interleukin-8 (IL-8) were measured in plasma by enzyme-linked immunosorbent assay. RESULTS: Compared with controls, sputum samples of ragpickers contained significantly increased numbers of alveolar macrophages, neutrophils, eosinophils and lymphocytes, suggesting airway inflammation. Circulating neutrophils and monocytes of the ragpickers overexpressed CD11b/CD18 and their platelets had upregulated surface expression of P-selectin, implying functional activation of these cells. In addition, plasma levels of IL-8 and TNF-alpha were significantly increased, indicating greater trafficking of leukocytes from circulation to the tissues. Multivariate logistic regression analysis demonstrated a positive association between the ragpicking profession and leukocyte activation after controlling for potential confounders. CONCLUSIONS: Ragpickers experience leukocyte and platelet activation and airway inflammation that could make them more vulnerable to tissue damage and cardiovascular diseases.
Asunto(s)
Bronquitis/fisiopatología , Leucocitos/metabolismo , Antígeno de Macrófago-1/metabolismo , Exposición Profesional/efectos adversos , Adulto , Bronquitis/etiología , Femenino , Residuos de Alimentos , Humanos , India , Antígeno de Macrófago-1/sangre , Persona de Mediana Edad , Selectina-P/sangre , Selectina-P/metabolismo , Pobreza , Factor de Necrosis Tumoral alfa/sangre , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Chronic arsenic exposure results in an increased oxidative stress and inflammation in the body. Glutamine (GLN) is an amino acid considered to have immunomodulatory effects and attenuate the inflammatory reaction. This study was designed to examine the effect of GLN supplementation on inflammatory-related leukocyte integrin expression and in vitro splenocyte cytokine production in mice exposed to arsenic. Mice were assigned to the control and experimental groups. The control group drank deionized water, whereas the experimental group drank deionized water containing 50 ppm of sodium arsenite. Each control and experimental group was further divided into 2 subgroups and fed diets for 5 weeks. One subgroup was fed a semipurified diet, whereas the other subgroup was fed a diet where part of the casein was replaced with GLN, which provided 25% of the total amino acid nitrogen. The results showed that plasma GLN levels of mice in the arsenic group were significantly lower than those in the control groups. Glutamine supplementation reversed the depletion of plasma GLN in the arsenic group. beta(2) intergins, including leukocyte function-associated antigen-1 and macrophage antigen-1 expressed by leukocytes, were significantly higher in the arsenic group than the control groups. Glutamine supplementation reduced leukocyte integrin expression in mice exposed to arsenic. There were no differences in interleukin 4, interleukin 6, interferon gamma, and tumor necrosis factor alpha production between the 2 arsenic groups when splenocytes were stimulated with mitogen. These results suggest that arsenic exposure results in depletion of plasma GLN and higher leukocyte integrin expression. Glutamine supplementation normalized the plasma GLN levels and reduced leukocyte leukocyte function-associated antigen-1 and macrophage antigen-1 expression. However, cytokine modulation may not be responsible for reducing leukocyte integrin expression in mice exposed to arsenic.
Asunto(s)
Arsénico/administración & dosificación , Glutamina/administración & dosificación , Antígeno-1 Asociado a Función de Linfocito/sangre , Antígeno de Macrófago-1/sangre , Animales , Arsénico/toxicidad , Células Cultivadas , Dieta , Citometría de Flujo , Glutamina/sangre , Leucocitos/química , Ratones , Ratones Endogámicos BALB C , Bazo/citologíaRESUMEN
Inflammation as well as platelet activation at the site of local vessel-wall injury plays an essential role in the mechanism of restenosis after Percutaneous coronary intervention (PCI). Platelet-derived microparticles (PDMPs) released from activated platelets are thought to play a role in the inflammatory process, possibly interacting with leukocyte integrin Mac-1. We serially measured circulating PDMPs, high-sensitive C-reactive protein (hs-CRP) and activated Mac-1 on the surface of neutrophils in 61 patients undergoing coronary stenting. PDMPs, hs-CRP and Mac-1 increased after coronary stenting in a time-dependent manner with the maximum response at 48 h in coronary sinus blood (PDMPs: 10.3+/-8.9-32.8+/-13.8 U/ml; P<0.001, hs-CRP: 0.27+/-0.23-1.46+/-0.99 mg/dl; P<0.001, activated Mac-1, 134+/-19% relative increase, P<0.001). PDMPs were correlated with hs-CRP (R=0.58, P<0.001) and the relative increase in Mac-1 (R=0.69, P<0.001) 48 h after coronary stenting. Multiple regression analysis showed that each of PDMPs (R=0.40, P<0.05), hs-CRP (R=0.33, P<0.05) and Mac-1 (R=0.48, P<0.01) was an independent predictor of the late lumen loss. Coronary stenting enhanced circulating PDMPs in association with an inflammatory response in the injured vessel wall. PDMPs may be a useful marker for evaluation of stent-induced inflammatory status and a powerful predictor of restenosis equivalent to activated Mac-1.