RESUMEN
BACKGROUND: Recently, we demonstrated that Astragalus polysaccharide (PG2), the active ingredient in dried roots of astragalus membranaceus, ameliorates cancer symptom clusters and improves quality of life (QoL) in patients with metastatic disease by modulating inflammatory cascade against the background roles of inflammatory cells, including macrophages, dendritic cells (DCs), and cytotoxic T lymphocytes (CTLs) in tumor initiation, metastasis, and progression. Nevertheless, the role of PG2 in the modulation of anticancer immunogenicity and therapeutic response remains relatively underexplored and unclear. PURPOSE: The present study investigates how and to what extent PG2 modulates cellular and biochemical components of the inflammatory cascade and enhances anticancer immunity, as well as the therapeutic implication of these bio-events in patients with lung cancer. METHODS AND RESULTS: Herein, we demonstrated that PG2 significantly increased the M1/M2 macrophage polarization ratio in non-small cell carcinoma (NSCLC) H441 and H1299 cells. This PG2-induced preferential pharmacologic up-regulation of tumoral M1 population in vitro positively correlated with the downregulation of tumor-promoting IL-6 and IL-10 expression in NSCLC cell-conditioned medium, with concomitant marked inhibition of cell proliferation, clonogenicity, and tumorsphere formation. Our ex vivo results, using clinical sample from our NSCLC cohort, demonstrated that PG2 also promoted the functional maturation of DCs with consequent enhancement of T cell-mediated anticancer immune responses. Consistent with the in vitro and ex vivo results, our in vivo studies showed that treatment with PG2 elicited significant time-dependent depletion of the tumor-associated M2 population, synergistically enhanced the anti-M2-based anticancer effect of cisplatin, and inhibited xenograft tumor growth in the NSCLC mice models. Moreover, in the presence of PG2, cisplatin-associated dyscrasia and weight-loss was markedly suppressed. CONCLUSION: These results do indicate a therapeutically-relevant role for PG2 in modulating the M1/M2 macrophage pool, facilitating DC maturation and synergistically enhancing the anticancer effect of conventional chemotherapeutic agent, cisplatin, thus laying the foundation for further exploration of the curative relevance of PG2 as surrogate immunotherapy and/or clinical feasibility of its use for maintenance therapy in patients with lung cancer.
Asunto(s)
Antineoplásicos/farmacología , Astragalus propinquus/química , Neoplasias Pulmonares/tratamiento farmacológico , Macrófagos/efectos de los fármacos , Polisacáridos/farmacología , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cisplatino/farmacología , Células Dendríticas/efectos de los fármacos , Galectina 3/efectos de los fármacos , Humanos , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Neoplasias Pulmonares/inmunología , Antígeno de Macrófago-1/efectos de los fármacos , Ratones , Calidad de Vida , Linfocitos T/efectos de los fármacosRESUMEN
Sepsis is a severe inflammatory condition associated with high mortality. Transmigration of neutrophils into tissues increases their lifespan to promote deleterious function. Junctional adhesion molecule-C (JAM-C) plays a pivotal role in neutrophil transmigration into tissues. We aim to study the role of JAM-C on the aging of neutrophils to cause sepsis-induced acute lung injury (ALI). Sepsis was induced in C57BL/6J mice by cecal ligation and puncture (CLP) and JAM-C expression in serum was assessed. Bone marrow-derived neutrophils (BMDN) were treated with recombinant mouse JAM-C (rmJAM-C) ex vivo and their viability was assessed. CLP-operated animals were administrated with either isotype IgG or anti-JAM-C Ab at a concentration of 3 mg/kg and after 20 h, aged neutrophils (CXCR4+ ) were assessed in blood and lungs and correlated with systemic injury and inflammatory markers. Soluble JAM-C level in serum was up-regulated during sepsis. Treatment with rmJAM-C inhibited BMDN apoptosis, thereby increasing their lifespan. CLP increased the frequencies of CXCR4+ neutrophils in blood and lungs, while treatment with anti-JAM-C Ab significantly reduced the frequencies of CXCR4+ aged neutrophils. Treatment with anti-JAM-C Ab significantly reduced systemic injury markers (alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase) as well as systemic and lung inflammatory cytokines (IL-6 and IL-1ß) and chemokine (macrophage inflammatory protein-2). The blockade of JAM-C improved lung histology and reduced neutrophil contents in lungs of septic mice. Thus, reduction of the pro-inflammatory aged neutrophils by blockade of JAM-C has a novel therapeutic potential in sepsis-induced ALI.
Asunto(s)
Lesión Pulmonar Aguda/tratamiento farmacológico , Moléculas de Adhesión Celular/antagonistas & inhibidores , Terapia Molecular Dirigida , Neutrófilos/inmunología , Sepsis/complicaciones , Lesión Pulmonar Aguda/etiología , Lesión Pulmonar Aguda/inmunología , Animales , Anticuerpos/uso terapéutico , Especificidad de Anticuerpos , Apoptosis , Moléculas de Adhesión Celular/sangre , Moléculas de Adhesión Celular/inmunología , Moléculas de Adhesión Celular/fisiología , Senescencia Celular , Citocinas/sangre , Inmunoglobulina G/farmacología , Inmunoglobulinas/sangre , Inmunoglobulinas/inmunología , Inmunoglobulinas/fisiología , Pulmón/inmunología , Pulmón/patología , Antígeno de Macrófago-1/efectos de los fármacos , Antígeno de Macrófago-1/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Neutrófilos/química , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores CXCR4/análisis , Proteínas Recombinantes/farmacología , Migración Transendotelial y Transepitelial/efectos de los fármacosAsunto(s)
Antígenos CD40/metabolismo , Ligando de CD40/antagonistas & inhibidores , Arterias Carótidas/efectos de los fármacos , Estenosis Carotídea/prevención & control , Antígeno de Macrófago-1/efectos de los fármacos , Neointima , Oligopéptidos/farmacología , Transducción de Señal/efectos de los fármacos , Animales , MasculinoRESUMEN
The co-stimulatory immune molecule CD40L figures prominently in a variety of inflammatory conditions including arterial disease. Recently, we made the surprising finding that CD40L mediates atherogenesis independently of its classic receptor CD40 via a novel interaction with the leukocyte integrin Mac-1. Here, we hypothesised that selective blockade of the CD40L-Mac-1 interaction may also retard restenosis. We induced neointima formation in C57/BL6 mice by ligation of the left carotid artery. Mice were randomised to daily intraperitoneal injections of either cM7, a small peptide selectively inhibiting the CD40L-Mac-1 interaction, scM7, a scrambled control peptide, or saline for 28 days. Interestingly, cM7-treated mice developed neointima of similar size compared with mice receiving the control peptide or saline as assessed by computer-assisted analysis of histological cross sections. These data demonstrate that the CD40L-Mac-1 interaction is not required for the development of restenosis. In contrast, CD40-deficient mice subjected to carotid ligation in parallel, developed significantly reduced neointimal lesions compared with respective wild-type controls (2872 ± 843 µm² vs 35469 ± 11870 µm²). Flow cytometry in CD40-deficient mice revealed reduced formation of platelet-granulocyte and platelet-inflammatory monocyte- aggregates. In vitro, supernatants of CD40-deficient platelet-leukocyte aggregates attenuated proliferation and increased apoptosis of smooth muscle cells. Unlike in the setting of atherosclerosis, CD40L mediates neointima formation via its classic receptor CD40 rather than via its recently described novel interaction with Mac-1. Therefore, selective targeting of CD40L-Mac-1 binding does not appear to be a favorable strategy to fight restenosis.
Asunto(s)
Antígenos CD40/metabolismo , Ligando de CD40/antagonistas & inhibidores , Arterias Carótidas/efectos de los fármacos , Estenosis Carotídea/prevención & control , Antígeno de Macrófago-1/efectos de los fármacos , Neointima , Oligopéptidos/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Apoptosis , Antígenos CD40/inmunología , Ligando de CD40/genética , Ligando de CD40/inmunología , Ligando de CD40/metabolismo , Arterias Carótidas/inmunología , Arterias Carótidas/metabolismo , Arterias Carótidas/patología , Estenosis Carotídea/inmunología , Estenosis Carotídea/metabolismo , Estenosis Carotídea/patología , Células Cultivadas , Modelos Animales de Enfermedad , Rodamiento de Leucocito/efectos de los fármacos , Antígeno de Macrófago-1/inmunología , Antígeno de Macrófago-1/metabolismo , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/metabolismo , Miocitos del Músculo Liso/patología , Activación Plaquetaria/efectos de los fármacos , RecurrenciaRESUMEN
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
BACKGROUND: Chewing of areca quid increases the prevalence of periodontal diseases. Areca nut extract (ANE) inhibits the phagocytic activity of human neutrophils. This in vitro study investigates the effects of ANE on complement- and antibody-opsonized phagocytosis by neutrophils. Expression of complement receptors, Fc receptors, and F-actin in ANE-treated neutrophils is also analyzed. METHODS: The viability of ANE-treated neutrophils was determined using the propidium iodide staining method. The possible effects of ANE on the expression of complement receptors and Fc receptors were examined using an immunofluorescence staining method followed by flow cytometry and confocal laser scanning microscopy. The phagocytic activity of neutrophils against complement or immunoglobulin (Ig)G-opsonized fluorescent beads was analyzed using flow cytometry. Expression of F-actin was determined using confocal laser scanning microscopy. RESULTS: ANE significantly inhibited the production of complement receptors (CR1, CR3, and CR4) and Fc receptors (FcγRII and FcγRIII) in a concentration-dependent manner. Treatment of neutrophils with ANE significantly impaired their ability to phagocytose fluorescent beads. ANE also inhibited phagocytosis of fluorescent beads that were opsonized by complement or IgG. Moreover, expression of F-actin was inhibited after ANE treatment. CONCLUSIONS: ANE inhibits the complement- and IgG-mediated neutrophil phagocytosis that may result from reduction of the expression of complement receptors, Fc receptors, and F-actin formation after ANE treatment. The findings suggest that areca nut chewing may jeopardize the defensive functions of neutrophils and affect periodontal health.
Asunto(s)
Areca , Neutrófilos/efectos de los fármacos , Nueces , Extractos Vegetales/farmacología , Receptores de Complemento/efectos de los fármacos , Receptores Fc/efectos de los fármacos , Actinas/efectos de los fármacos , Adulto , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Colorantes , Complemento C1/efectos de los fármacos , Femenino , Citometría de Flujo , Técnica del Anticuerpo Fluorescente Directa , Humanos , Integrina alfaXbeta2/efectos de los fármacos , Antígeno de Macrófago-1/efectos de los fármacos , Masculino , Microscopía Confocal , Microesferas , Neutrófilos/inmunología , Fagocitosis/efectos de los fármacos , Propidio , Receptores de IgG/efectos de los fármacos , Adulto JovenRESUMEN
Interleukin-1ß (IL-1ß) is a proinflammatory cytokine and a therapeutic target in several chronic autoimmune states. Monocytes and macrophages are the major sources of IL-1ß. IL-1ß production by these cells requires Toll-like receptor (TLR) and adenosine triphosphate (ATP)-mediated P2X purinoceptor 7 (P2X7) signals, which together activate the inflammasome. However, how TLR signals and ATP availability are regulated during monocyte activation is unclear and the involvement of another danger signal system has been proposed. Here, we demonstrate that both lipopolysaccharide (LPS) and the anaphylatoxin C3a are needed for IL-1ß production in human macrophages and dendritic cells, while in monocytes, C3a enhanced the secretion of LPS-induced IL-1ß. C3a and LPS-stimulated monocytes increased T helper 17 (Th17) cell induction in vitro, and human rejecting, but not nonrejecting, kidney transplant biopsies were characterized by local generation of C3a and monocyte and Th17 cell infiltration. Mechanistically, C3a drives IL-1ß production in monocytes by controlling the release of intracellular ATP into the extracellular space via regulation of as-yet unidentified ATP-releasing channels in an extracellular signal-regulated kinase 1/2-dependent fashion. These data define a novel function for complement in inflammasome activation in monocytes and suggest that C3aR-mediated signaling is a vital component of the IL-1ß-Th17 axis.
Asunto(s)
Adenosina Trifosfato/metabolismo , Proteínas Portadoras/fisiología , Complemento C3/fisiología , Inflamasomas/fisiología , Interleucina-1beta/metabolismo , Monocitos/metabolismo , Células Cultivadas , Complemento C3/agonistas , Citocinas/biosíntesis , Citocinas/genética , Células Dendríticas/metabolismo , Activación Enzimática , Rechazo de Injerto/inmunología , Rechazo de Injerto/patología , Humanos , Trasplante de Riñón , Lipopolisacáridos/farmacología , Sistema de Señalización de MAP Quinasas , Antígeno de Macrófago-1/efectos de los fármacos , Antígeno de Macrófago-1/fisiología , Macrófagos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR , Receptores Purinérgicos P2X7/fisiología , Proteínas Recombinantes/farmacología , Células Th17/metabolismo , Receptores Toll-Like/fisiologíaRESUMEN
Magnetic nanoparticles surface-covered with meso-2,3-dimercaptosuccinic acid (MNPs-DMSA) constitute a promising approach for tissue- and cell-targeted delivery of therapeutic drugs in the lung. However, they can also induce a transient transendothelial migration of leukocytes in the organ as a side effect after endovenous administration of MNPs-DMSA. We demonstrated that monocytes/macrophages constitute the main subpopulation of leukocytes involved in this process. Our recent research found that MNPs-DMSA up-regulated the mRNA expression of E-, L- and P-selectin and macrophage-1 antigen, and increased concentration of tumor necrosis factor-α in lung, in a time dependent manner. The critical relevance of the ß2 integrin-dependent pathway in leukocyte transmigration elicited by MNPs-DMSA was demonstrated by use of knockout mice. Our work characterizes mechanisms of the pro-inflammatory effects of MNPs-DMSA in the lung, and identifies ß2 integrin-targeted interventions as promising strategies to reduce pulmonary side effects of MNPs-DMSA during biomedical applications. In addition, MNPs-DMSA could be used as modulators of lung immune response.
Asunto(s)
Leucocitos/efectos de los fármacos , Pulmón/efectos de los fármacos , Nanopartículas de Magnetita/administración & dosificación , Succímero/administración & dosificación , Migración Transendotelial y Transepitelial/efectos de los fármacos , Animales , Antígenos CD18/metabolismo , Leucocitos/metabolismo , Antígeno de Macrófago-1/efectos de los fármacos , Antígeno de Macrófago-1/metabolismo , Macrófagos/efectos de los fármacos , Ratones , Selectina-P/efectos de los fármacos , Selectina-P/metabolismo , Succímero/química , Factor de Necrosis Tumoral alfa/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Intracellular signals associated with or triggered by integrin ligation can control cell survival, differentiation, proliferation, and migration. Despite accumulating evidence that conformational changes regulate integrin affinity to its ligands, how integrin structure regulates signal transmission from the outside to the inside of the cell remains elusive. Using fluorescence resonance energy transfer, we addressed whether conformational changes in integrin Mac-1 are sufficient to transmit outside-in signals in human neutrophils. Mac-1 conformational activation induced by ligand occupancy or activating Ab binding, but not integrin clustering, triggered similar patterns of intracellular protein tyrosine phosphorylation, including Akt phosphorylation, and inhibited spontaneous neutrophil apoptosis, indicating that global conformational changes are critical for Mac-1-dependent outside-in signal transduction. In neutrophils and myeloid K562 cells, ligand ICAM-1 or activating Ab binding promoted switchblade-like extension of the Mac-1 extracellular domain and separation of the alpha(M) and beta(2) subunit cytoplasmic tails, two structural hallmarks of integrin activation. These data suggest the primacy of global conformational changes in the generation of Mac-1 outside-in signals.
Asunto(s)
Leucocitos Mononucleares/inmunología , Antígeno de Macrófago-1/inmunología , Neutrófilos/inmunología , Transducción de Señal/inmunología , Anticuerpos Monoclonales/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/inmunología , Carcinógenos/farmacología , Adhesión Celular/efectos de los fármacos , Adhesión Celular/inmunología , Línea Celular Tumoral , Humanos , Factores Inmunológicos/farmacología , Molécula 1 de Adhesión Intercelular/inmunología , Molécula 1 de Adhesión Intercelular/metabolismo , Interleucina-8/farmacología , Leucemia/inmunología , Leucemia/metabolismo , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Antígeno de Macrófago-1/efectos de los fármacos , Antígeno de Macrófago-1/metabolismo , N-Formilmetionina Leucil-Fenilalanina/farmacología , Neutrófilos/efectos de los fármacos , Neutrófilos/metabolismo , Estructura Terciaria de Proteína/efectos de los fármacos , Estructura Terciaria de Proteína/fisiología , Transducción de Señal/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
LPS activates microglia, which are normally maintained in a quiescent state by CD200-CD200 receptor (CD200R) interaction. MAC-1 (a microglia marker) mRNA expression was increased in mice brains up to 1 year post LPS administration (i.p.). Minocycline treatment did not prevent LPS (5 mg/kg)-induced increase in MAC-1 mRNA but reduced that induced by 0.1 mg/kg LPS. CD200R mRNA decreased starting at 4 h, whereas CD200 mRNA increased at 4 h and decreased at 1 year post LPS inoculation. Thus, LPS-induced changes in CD200-CD200R equilibrium might keep microglia chronically activated. Minocycline does not effectively inhibit microglia activation induced by high-dose LPS.
Asunto(s)
Antígenos CD/metabolismo , Encéfalo/inmunología , Antígeno de Macrófago-1/metabolismo , Glicoproteínas de Membrana/metabolismo , Microglía/inmunología , Animales , Antibacterianos/farmacología , Antígenos CD/efectos de los fármacos , Encéfalo/citología , Encéfalo/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Lipopolisacáridos , Antígeno de Macrófago-1/efectos de los fármacos , Glicoproteínas de Membrana/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Microglía/efectos de los fármacos , Minociclina/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de TiempoRESUMEN
Three series of non-steroidal anti-inflammatory drugs (NSAIDs) inhibiting the cyclooxygenase/5-lipoxygenase (COX/5-LOX) pathways as such as formation of hydroxyl radicals and adhesion were prepared: 4,5-diaryl isothiazoles, 4,5-diaryl 3H-1,2-dithiole-3-thiones and 4,5-diaryl 3H-1,2-dithiole-3-ones. The aim of the present study was to develop substances which can intervene into the inflammatory processes via different mechanisms of action as multiple target non-steroidal anti-inflammatory drugs (MTNSAIDs) with increased anti-inflammatory potential. The current lead 11a was evaluated in COX-1/2, 5-LOX and (*)OH scavenging in vitro assays and in a static adhesion assay where it proved to inhibit adhesion. Moreover, 11a treatment attenuated expression of macrophage adhesion molecule-1 (Mac-1) on extravasated polymorphonuclear leukocytes (PMNs) which indicates that the activation was reduced. The assays used are predictive for the in vivo efficacy of test compounds as shown for 11a in a peritonitis model of acute inflammation in mice. Thus, the novel 5-LOX/COX and (*)OH inhibitor 11a possesses anti-inflammatory activity that, in addition to COX/5-LOX inhibition, implicates effects on leukocyte-endothelial interactions.
Asunto(s)
Antiinflamatorios no Esteroideos/química , Inhibidores de la Ciclooxigenasa/química , Inhibidores de la Lipooxigenasa , Compuestos de Sulfhidrilo/química , Tiazoles/química , Animales , Antiinflamatorios no Esteroideos/farmacología , Adhesión Celular/efectos de los fármacos , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa/farmacología , Depuradores de Radicales Libres , Radical Hidroxilo , Antígeno de Macrófago-1/análisis , Antígeno de Macrófago-1/efectos de los fármacos , Ratones , Neutrófilos/efectos de los fármacos , Peritonitis/tratamiento farmacológico , Compuestos de Sulfhidrilo/farmacología , Tiazoles/farmacologíaRESUMEN
This study was conducted to investigate putative antagonism of integrin receptors alphaMbeta2 and alphaLbeta2 by a novel coumarin derivative (BOL-303225-A), its efficacy in-vivo after retinal ischaemia-reperfusion injury, and its bioavailability in rat plasma. A cellular adhesion assay in Jurkat and U937 cells, and a flow cytometry assay with an antibody against the beta2 subunit were conducted. BOL-303225-A bioavailability in rat plasma and the retinal levels of myeloperoxidase (MPO) after ischaemia-reperfusion injury were evaluated after oral administration (10 mg kg(-1)). In-vitro cell viability assays revealed no cytotoxicity for BOL-303225-A over a wide dose range, and IC50 values of 32.3 +/- 1.5 muM and 84.95 +/- 2.3 muM were found for Jurkat and U937 cells, respectively. The drug showed specific binding to the alphaMbeta2 and alphaLbeta2 integrin receptors expressed by U937 and Jurkat cells, respectively, producing a fluorescence shift towards lower values in a concentration-dependent manner. The pharmacokinetic profile of BOL-303225-A exhibited rapid absorption following oral administration in the rat. A significant reduction of retinal MPO levels was observed in drug-treated rats. This study demonstrated that BOL-303225-A acts as an antagonist of the integrin alphaLbeta2 and alphaMbeta2 receptors, suggesting that this drug could be used for ocular diseases such as diabetic retinopathy.
Asunto(s)
Antiinflamatorios/farmacología , Cumarinas/farmacología , Antígeno-1 Asociado a Función de Linfocito/efectos de los fármacos , Antígeno de Macrófago-1/efectos de los fármacos , Administración Oral , Animales , Antiinflamatorios/administración & dosificación , Antiinflamatorios/farmacocinética , Disponibilidad Biológica , Adhesión Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cumarinas/administración & dosificación , Cumarinas/farmacocinética , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Humanos , Células Jurkat , Antígeno-1 Asociado a Función de Linfocito/metabolismo , Antígeno de Macrófago-1/metabolismo , Masculino , Peroxidasa/efectos de los fármacos , Peroxidasa/metabolismo , Unión Proteica , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/patología , Enfermedades de la Retina/tratamiento farmacológico , Enfermedades de la Retina/patología , Células U937RESUMEN
The molecular and structural basis of anesthetic interactions with conformations and functionalities of cell surface receptors remains to be elucidated. We have demonstrated that the widely used volatile anesthetic isoflurane blocks the activation-dependent conformational conversion of integrin lymphocyte function associated antigen-1 (LFA-1), the major leukocyte cell adhesion molecule, to a high-affinity configuration. Perturbation of LFA-1 activation by isoflurane at clinically relevant concentrations leads to the inhibition of T-cell interactions with target cells as well as ligand-triggered intracellular signaling. Nuclear magnetic resonance spectroscopy reveals that isoflurane binds within a cavity in the LFA-1 ligand-binding domain, which is a previously identified drug-binding site for allosteric small-molecule antagonists that stabilize LFA-1 in a low-affinity conformation. These results provide a potential mechanism for the immunomodulatory properties of isoflurane.
Asunto(s)
Sitio Alostérico/efectos de los fármacos , Anestésicos por Inhalación/química , Anestésicos por Inhalación/farmacología , Isoflurano/química , Isoflurano/farmacología , Antígeno-1 Asociado a Función de Linfocito/química , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Células Jurkat , Células K562 , Antígeno-1 Asociado a Función de Linfocito/metabolismo , Antígeno de Macrófago-1/efectos de los fármacos , Antígeno de Macrófago-1/metabolismo , Modelos Biológicos , Resonancia Magnética Nuclear Biomolecular , Unión Proteica , Conformación Proteica/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
C3 binding glycoprotein (C3bgp) is immunomodulating molecule isolated from the plant Cuscuta europea. When neutrophils were incubated with C3bgp the subsequent binding of anti-CD11b mAb became significantly higher. C3bgp induced moderate TNF-alpha production in human PBMC and primary monocytes. This production was significantly inhibited by the specific inhibitors of JNK and p38 MAPKs. The inhibition of JNK reduced PBMC viability. We concluded that: (i) C3bgp utilized CD11b polypeptide chain of CR3 and mediated a part of its immunomodulatory properties by activation of JNK and p38 and (ii) PBMC viability at in vitro conditions depends of JNK signal transduction pathway activation.
Asunto(s)
Proteínas Portadoras/farmacología , Glicoproteínas/farmacología , Factores Inmunológicos , Antígeno de Macrófago-1/efectos de los fármacos , Proteínas Quinasas Activadas por Mitógenos/fisiología , Transducción de Señal/efectos de los fármacos , Antígeno CD11b/metabolismo , Supervivencia Celular/efectos de los fármacos , Ensayo de Inmunoadsorción Enzimática , Humanos , Lipopolisacáridos/farmacología , MAP Quinasa Quinasa 4/metabolismo , Neutrófilos/efectos de los fármacos , Neutrófilos/metabolismo , Fagocitos/efectos de los fármacos , Factor de Necrosis Tumoral alfa/biosíntesis , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Experimental toxocariasis was used as a model of eosinophil migration. Mice inoculated with 200 Toxocara canis eggs were treated with the leukotriene inhibitor MK886 (1 mg/kg/day). Eosinophils were counted in peripheral blood (PB), peritoneal cavity (PC) and bronchoalveolar lavage fluid (BALF) samples on post-infection days 3, 6, 12, 18, 24 and 36. Eosinophil expression of Mac-1 and VLA-4 was analysed in PB and PC samples. We found that T. canis infection induced systemic eosinophilia from post-infection day 3, peaking on days 6, 12 and 24 in PB, PC and BALF samples respectively. Eosinophilia was more pronounced in PB and PC samples than in BALF samples, and MK886 downregulated eosinophilia to varying degrees in the different sample types. In PB and PC samples, T. canis infection caused early upregulation of Mac-1 with late changes in the VLA-4 profile, whereas MK886 had opposite effects. The distinct time-dependent eosinophilia peaks and differential involvement of leukotrienes in integrin expression demonstrate that, despite the systemic eosinophilia triggered by T. canis infection, inflammatory responses vary by compartment.
Asunto(s)
Eosinofilia/tratamiento farmacológico , Eosinófilos/efectos de los fármacos , Indoles/uso terapéutico , Inhibidores de la Lipooxigenasa/uso terapéutico , Toxocariasis/tratamiento farmacológico , Animales , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Movimiento Celular/efectos de los fármacos , Movimiento Celular/inmunología , Eosinofilia/inmunología , Eosinófilos/inmunología , Femenino , Citometría de Flujo , Integrina alfa4beta1/biosíntesis , Integrina alfa4beta1/efectos de los fármacos , Antígeno de Macrófago-1/biosíntesis , Antígeno de Macrófago-1/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Fenotipo , Toxocariasis/inmunologíaRESUMEN
The platelet glycoprotein (GP) IIb/IIIa inhibitors differ markedly in their pharmacokinetics, pharmacodynamics, and differential receptor affinities. Abciximab and the small-molecule GP IIb/IIIa inhibitors (eptifibatide, tirofiban) have separate, distinct binding sites on the GP IIb/IIIa receptor complex. The affinity of abciximab for the platelet GP IIb/IIIa integrin receptor, together with non-platelet-receptor mediated effects achieved through its affinity for the aVb3 and CD11b/18 receptors, most likely contribute to the clinical benefit associated with the use of abciximab as adjunctive pharmacotherapy during primary percutaneous coronary intervention for the treatment of ST-elevation acute myocardial infarction.
Asunto(s)
Angioplastia Coronaria con Balón/efectos adversos , Antiinflamatorios/uso terapéutico , Plaquetas/efectos de los fármacos , Circulación Coronaria/efectos de los fármacos , Infarto del Miocardio/terapia , Inhibidores de Agregación Plaquetaria/uso terapéutico , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/antagonistas & inhibidores , Vasculitis/prevención & control , Abciximab , Antiinflamatorios/farmacología , Anticuerpos Monoclonales/uso terapéutico , Plaquetas/metabolismo , Humanos , Fragmentos Fab de Inmunoglobulinas/uso terapéutico , Integrina alfaVbeta3/antagonistas & inhibidores , Antígeno de Macrófago-1/efectos de los fármacos , Microcirculación/efectos de los fármacos , Infarto del Miocardio/sangre , Infarto del Miocardio/tratamiento farmacológico , Infarto del Miocardio/fisiopatología , Inhibidores de Agregación Plaquetaria/farmacología , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , Vasculitis/etiologíaRESUMEN
Complement-receptor-3 (CR3/MAC-1), scavenger-receptor-AI/II (SRAI/II), and Fcgamma-receptor (FcgammaR) can mediate myelin phagocytosis in macrophages and microglia. Paradoxically, after injury to CNS axons these receptors are expressed but myelin is not phagocytosed, suggesting that phagocytosis is subject to regulation between efficient and inefficient states. In the present work, we focus on CR3/MAC-1 and SRAI/II-mediated myelin phagocytosis. Phagocytosis by CR3/MAC-1 and SRAI/II was inhibited by cPKC inhibitor Go-6976, general-PKC inhibitors Ro-318220 and calphostin-C, and BAPTA/AM, which chelates intracellular Ca2+ required for cPKC activation. Signaling/activation by cPKC are thus suggested. PMA, which mimics diacylglycerol (DAG) as an activator of cPKC, novel-PKC (nPKC), and non-PKC DAG-driven molecule(s), produced a dose-dependent dual effect on phagocytosis by CR3/MAC-1 and SRAI/II, i.e., augmentation at low concentrations and inhibition at high concentrations. Inhibition of phagocytosis by CR3/MAC-1 was enhanced by combining inhibiting concentrations of PMA with PKC inhibitors Go-6976 or Ro-318220, suggesting inhibition by PMA/DAG-driven non-PKC molecule(s). In contrast, inhibition of phagocytosis by SRAI/II was enhanced by combining inhibiting concentrations of PMA with cPKC inhibitor Go-6976 but not with general-PKC inhibitor Ro-318220, suggesting inhibition by nPKC. Phagocytosis by CR3/MAC-1 and SRAI/II was further inhibited by PI3K inhibitors wortmannin and LY-294002 and PLCgamma inhibitor U-73122. Altogether, our observations suggest that CR3/MAC-1 and SRAI/II-mediated myelin phagocytosis share activation by PI3K, PLCgamma and cPKC. The two differ, however, in that non-PKC DAG-driven molecule(s) inhibit CR3/MAC-1-mediated phagocytosis, whereas nPKC inhibit SRAI/II-mediated phagocytosis. Each of these signaling steps may be targeted for regulating CR3/MAC-1 and/or SRAI/II-mediated phagocytosis between efficient and inefficient states.
Asunto(s)
Antígeno de Macrófago-1/efectos de los fármacos , Vaina de Mielina/inmunología , Fagocitosis/fisiología , Fosfatidilinositol 3-Quinasas/farmacología , Fosfolipasa C gamma/farmacología , Proteína Quinasa C/farmacología , Receptores Depuradores de Clase A/antagonistas & inhibidores , Animales , Quelantes/farmacología , Relación Dosis-Respuesta a Droga , Ácido Egtácico/análogos & derivados , Ácido Egtácico/farmacología , Activación Enzimática/fisiología , Ensayo de Inmunoadsorción Enzimática , Immunoblotting , Inmunoprecipitación , Isoenzimas/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Microglía/efectos de los fármacos , Microglía/fisiología , Receptores de Droga , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Acetato de Tetradecanoilforbol/farmacología , Tioglicolatos/farmacología , Tirosina/metabolismoAsunto(s)
Anticuerpos Monoclonales/uso terapéutico , Antineoplásicos/uso terapéutico , Factores Inmunológicos/uso terapéutico , Linfoma de Células B/tratamiento farmacológico , Animales , Anticuerpos Monoclonales de Origen Murino , Humanos , Linfoma de Células B/inmunología , Antígeno de Macrófago-1/efectos de los fármacos , Antígeno de Macrófago-1/inmunología , RituximabRESUMEN
Two subfamilies of the polar glycopeptidolipids (GPLs) located on the surface of Mycobacterium smegmatis, along with unknown phospholipids, were recently shown to participate in the nonopsonic phagocytosis of mycobacteria by human macrophages (Villeneuve, C., G. Etienne, V. Abadie, H. Montrozier, C. Bordier, F. Laval, M. Daffe, I. Maridonneau-Parini, and C. Astarie-Dequeker. 2003. Surface-exposed glycopeptidolipids of Mycobacterium smegmatis specifically inhibit the phagocytosis of mycobacteria by human macrophages. Identification of a novel family of glycopeptidolipids. J. Biol. Chem. 278: 51291-51300). As demonstrated herein, a phospholipid mixture that derived from the methanol-insoluble fraction inhibited the phagocytosis of M. smegmatis. Inhibition was essentially attributable to phosphatidylinositol mannosides (PIMs), namely PIM2 and PIM6, because the purified phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylinositol were inactive. This was further confirmed using purified PIM2 and PIM6 from M. bovis BCG that decreased by half the internalization of M. smegmatis. Both compounds also inhibited the uptake of M. tuberculosis and M. avium but had no effect on the internalization of zymosan used as a control particle of the phagocytic process. When coated on latex beads, PIM2 and polar GPL (GPL III) favored the particle entry through complement receptor 3. GPL III, but not PIM2, also directed particle entry through the mannose receptor. Therefore, surface-exposed mycobacterial PIM and polar GPL participate in the receptor-dependent internalization of mycobacteria in human macrophages.
Asunto(s)
Glucolípidos/farmacología , Antígeno de Macrófago-1/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/microbiología , Mycobacterium smegmatis/química , Fagocitosis/efectos de los fármacos , Secuencia de Carbohidratos , Células Cultivadas , Glucolípidos/química , Glucolípidos/aislamiento & purificación , Humanos , Antígeno de Macrófago-1/efectos de los fármacos , Macrófagos/inmunología , Espectrometría de Masas , Datos de Secuencia Molecular , Estructura Molecular , Mycobacterium bovis/química , Mycobacterium smegmatis/efectos de los fármacos , Mycobacterium smegmatis/fisiología , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/fisiología , Fosfatidilinositoles/química , Fosfatidilinositoles/farmacologíaRESUMEN
OBJECTIVES: The aim of this study was to confirm clinically a hypothesis that cilostazol inhibits leukocyte Mac-1, leading to prevention of post-stent restenosis. BACKGROUND: The platelet phosphodiesterase III inhibitor called cilostazol also inhibits alpha-granule release of P-selectin in platelets. The P-selectin-mediated platelet-leukocyte interaction promotes activation and upregulation of leukocyte Mac-1 after coronary stenting, which plays a key role on the mechanism of restenosis. Thus, cilostazol's potential inhibition of this process may lead to prevention of restenosis. METHODS: Using flow cytometric analysis of whole blood obtained from the coronary sinus, the expression of platelet membrane glycoproteins and neutrophil adhesion molecules was observed in 70 consecutive patients undergoing coronary stenting. The patients were randomly assigned to either a cilostazol or ticlopidine group before stent placement. RESULTS: The restenosis rate was lower (15% vs. 31%, p < 0.05) in the cilostazol group (n = 34) than in the ticlopidine group (n = 32). A stent-induced increase in platelet P-selectin (CD62P) expression and an increase in neutrophil Mac-1 (CD11b) expression were suppressed in the cilostazol group compared with the ticlopidine group. Angiographic late lumen loss was correlated with the relative changes in platelet P-selectin and neutrophil Mac-1 at 48 h after coronary stenting. CONCLUSIONS: Cilostazol may have effects on suppression of P-selectin-mediated platelet activation, platelet-leukocyte interaction, and subsequent Mac-1-mediated leukocyte activation, which might lead to a reduced restenosis rate after coronary stent implantation.