RESUMEN
Immune checkpoint inhibitors (ICIs) have completely reshaped the treatment of many malignancies, with remarkable improvements in survival outcomes. In ovarian cancer (OC), however, this emerging class of drugs has not yet found a favorable use due to results from phase I and II studies, which have not suggested a substantial antitumoral activity of these agents when administered as monotherapy. Robust preclinical data seem to suggest that the combination ICIs with poly(ADP-ribose) polymerase (PARP) inhibitors (PARPis) may result in a synergistic activity; furthermore, data from phase II clinical studies, evaluating this combination, have shown encouraging outcomes especially for those OC patients not suitable for platinum retreatment. While waiting for ongoing phase III clinical trial results, which will clarify the role of ICIs in combination with PARPis in the newly diagnosed OC, this review aims to summarize the preclinical data and clinical evidence available to date.
Asunto(s)
Neoplasias Ováricas , Inhibidores de Poli(ADP-Ribosa) Polimerasas , Femenino , Humanos , Inhibidores de Puntos de Control InmunológicoRESUMEN
Integrin α4ß7 mediates the trafficking of leukocytes, including CD4+ T cells, to lymphoid tissues in the gut. Virus mediated damage to the gut is implicated in HIV and SIV mediated chronic immune activation and leads to irreversible damage to the immune system. We employed an immuno-PET/CT imaging technique to evaluate the impact of an anti-integrin α4ß7 mAb alone or in combination with ART, on the distribution of both SIV infected cells and CD4+ cells in rhesus macaques infected with SIV. We determined that α4ß7 mAb reduced viral antigen in an array of tissues of the lung, spleen, axillary, and inguinal lymph nodes. These sites are not directly linked to α4ß7 mediated homing; however, the most pronounced reduction in viral load was observed in the colon. Despite this reduction, α4ß7 mAb treatment did not prevent an apparent depletion of CD4+ T cells in gut in the acute phase of infection that is characteristic of HIV/SIV infection. However, α4ß7 mAb appeared to facilitate the preservation or restoration of CD4+ T cells in gut tissues at later stages of infection. Since damage to the gut is believed to play a central role in HIV pathogenesis, these results support further evaluation of α4ß7 antagonists in the study and treatment of HIV disease.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Colon/virología , Infecciones por VIH/inmunología , VIH-1/fisiología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/fisiología , Animales , Anticuerpos Monoclonales/metabolismo , Proteínas de la Membrana Bacteriana Externa , Linfocitos T CD4-Positivos/virología , Supervivencia Celular , Supresión Clonal , Modelos Animales de Enfermedad , Humanos , Integrinas/inmunología , Macaca , Receptores de Superficie Celular , Carga ViralRESUMEN
Thrombin (THR) plays a key role in the brain under physiological and pathological conditions. Several of the biological activities of thrombin have been shown to be mainly driven through activation of protease-activated receptor-1 (PAR-1)-type thrombin receptor. Here we have studied the effect of THR and PAR-1-activating peptide (PAR1-AP), SFLLRN, on cytokine-induced expression of inducible nitric oxide (iNOS), a prominent marker of astroglial activation using the rat C6 glioma cells. In this cell line, THR (1-10 U/mL) and PAR1-AP (1-100 microM) induced a significant concentration-dependent increase both of IFN-gamma- (250 U/mL) or TNF-alpha- (500 U/mL) induced NO release. The observed increase of NO production was related to an enhancement of iNOS expression as measured in cell lysates prepared from different treatments by using SDS-PAGE followed by western blot analysis. The effect of THR, but not that of PAR1-AP, was significantly inhibited by hirulog(TM) (60 microg/mL), a specific and stochiometric THR inhibitor or by cathepsin-G (40 mU/mL), an inhibitor of PAR-1. In conclusion our data suggest a role for THR through activation of PAR-1 in the induction of astroglial iNOS, and further support the hypothesis that THR may function as an important pathophysiological modulator of the inflammatory response.
Asunto(s)
Hemostáticos/farmacología , Interferón gamma/farmacología , Óxido Nítrico Sintasa/metabolismo , Fragmentos de Péptidos/farmacología , Trombina/farmacología , Factor de Necrosis Tumoral alfa/farmacología , Animales , Astrocitos/citología , Astrocitos/efectos de los fármacos , Astrocitos/enzimología , Catepsina G , Catepsinas/farmacología , División Celular/efectos de los fármacos , Sinergismo Farmacológico , Glioma , Óxido Nítrico Sintasa de Tipo II , Nitritos/metabolismo , Ratas , Serina Endopeptidasas , Células Tumorales CultivadasRESUMEN
BACKGROUND AND AIM: Platelet aggregation is involved in atherosclerosis and pharmacological inhibition of platelet activity may reduce the risk of coronary thrombosis and myocardial infarction. Red wine polyphenols may reduce platelet aggregability. This study evaluates the effect of de-alcoholated red wine (DRW) and its phenolic fractions on rat platelet aggregation and cyclic AMP (c-AMP) content. METHODS AND RESULTS: DRW was fractionated into four classes of phenolic compounds: phenolic acids (fraction 1), procyanidins, catechins and monomeric anthocyanidins (fraction 2), flavonols and resveratrol (fraction 3) and polymeric anthocyanidins (fraction 4). The effect of each fraction on ADP-induced rat platelet aggregation and c-AMP content was compared with that of DRW and pure phenolic compounds (quercetin, catechin, resveratrol, caffeic acid). DRW completely inhibited ADP-induced platelet aggregation. Fraction 2 also showed a significant anti-aggregating activity, whereas the effects of fractions 3 and 4 and the pure phenolics were not significant. A significant increase in platelet c-AMP content was observed after the addition of DRW and fraction 2. CONCLUSIONS: Our data indicate that DRW and its catechin-anthocyanidin fraction exert a significant effect on platelet aggregation in vitro, perhaps by enhancing platelet c-AMP levels.
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Enfermedades Cardiovasculares/prevención & control , AMP Cíclico/metabolismo , Flavonoides , Fenoles/farmacología , Agregación Plaquetaria/efectos de los fármacos , Polímeros/farmacología , Vino/análisis , Animales , Enfermedades Cardiovasculares/sangre , Etanol , Masculino , Fenoles/química , Inhibidores de Agregación Plaquetaria , Polímeros/química , Polifenoles , Ratas , Ratas WistarAsunto(s)
Presión Sanguínea/efectos de los fármacos , Oligopéptidos/farmacología , Receptores de Trombina/metabolismo , Animales , Clorisondamina/farmacología , Bloqueadores Ganglionares/farmacología , Hipotensión/inducido químicamente , Modelos Biológicos , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/farmacología , Fragmentos de Péptidos/farmacología , Ratas , Receptor PAR-1 , Receptor PAR-2RESUMEN
A phytochemical investigation of the extracts obtained from bulbs of leek. Allium porrum L. has led to the isolation of five flavonoid glycosides based on the kaempferol aglycone. Two of them are new compounds and have been identified as kaempferol 3-O-[2-O-(trans-3-methoxy-4-hydroxycinnamoyl)-beta-D-galactopyranosyl]-(1-->4)-O-beta-D-glucopyranoside, and kaempferol 3-O-[2-O-(trans-3-methoxy-4-hydroxycinnamoyl)-beta-D-glucopyranosyl]-(1-->6)-O-beta-D-glucopyranoside, on the basis of spectroscopic methods, including 2D NMR. The isolated compounds have been evaluated for their human platelet anti-aggregation activity.
Asunto(s)
Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Quempferoles , Cebollas/química , Agregación Plaquetaria/efectos de los fármacos , Quercetina/aislamiento & purificación , Flavonoides/química , Humanos , Espectroscopía de Resonancia Magnética , Quercetina/análogos & derivados , Quercetina/química , Quercetina/farmacologíaRESUMEN
1. Protease activated receptor-2 (PAR2) is a seven transmembrane domain G protein coupled receptor proteolytically activated. PAR2, together with other PARs, can be also activated by peptides mimicking the sequence of the receptor tethered ligand. We have evaluated the effect of systemic administration of a peptide activating PAR2 (PAR2-AP, SLIGRL) on histamine-induced increase in lung resistances in the guinea-pig. 2. Intravenous administration of PAR2-AP (1 mg kg(-1)) significantly inhibited histamine-induced increase in lung resistance in a time-dependent fashion that was not abolished by indomethacin or vagotomy. 3. Bronchoprotective effect of PAR2-AP was not reversed by the cyclo-oxygenase inhibitor, indomethacin, the nitric oxide synthetase inhibitor, L-NAME, nor by the non-selective beta-antagonist, propranolol. 4. Indomethacin augmented the bronchoconstriction to histamine which was inhibited by PAR2-AP. Furthermore, in vagotomized animals, the bronchial hyper-responsiveness to histamine was significantly reduced, and in these circumstances, PAR2-AP still retained the capacity to provide bronchoprotection against histamine. 5. PAR2-AP also produced a modest reduction in histamine-induced protein leakage in trachea and upper bronchi. 6. Our results indicated that PAR2 might have a bronchoprotective role in the guinea-pig in vivo independent of prostaglandin or nitric oxide release.
Asunto(s)
Broncoconstricción/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Sustancias Protectoras/farmacología , Receptores de Trombina/metabolismo , Análisis de Varianza , Animales , Espasmo Bronquial/inducido químicamente , Espasmo Bronquial/prevención & control , Modelos Animales de Enfermedad , Cobayas , Histamina , Fragmentos de Péptidos/uso terapéutico , Permeabilidad , Sustancias Protectoras/uso terapéutico , Receptor PAR-2 , VagotomíaRESUMEN
Histamine, vascular endothelial growth factor, acetylcholine, oestrogen as well as fluid shear stress activates a mechanism that recruits heat shock protein 90 to the endothelial nitric oxide synthase. The interaction between Hsp90 and eNOS enhances the activation of the enzyme in cells and in intact blood vessels leading to NO production. Intraplantar administration of carrageenan (50 microl paw(-1)) to mice causes an oedema lasting 72 h. Geldanamycin (0.1, 0.3, 1 mg kg(-1)), a specific inhibitor of Hsp-90, that inhibits endothelium-dependent relaxations of the rat aorta, mesentery and middle artery inhibits carrageenan-induced mouse paw oedema in a dose dependent manner. Co-administration to mice of dexamethasone (1 mg kg(-1)) with geldanamycin (0.3 mg kg(-1)) at anti-inflammatory dose causes a loss of the total anti-inflammatory effect of each agent alone. RU 486 (10 mg kg(-1)), a well known glucocorticoid receptorial antagonist, does not inhibit oedema formation but prevents the anti-inflammatory action of dexamethasone (1 mg kg(-1)). Similarly, RU 486 prevents the anti-inflammatory action of geldanamycin (0.3 mg kg(-1)). In conclusion we have described for the first time that geldanamycin, an inhibitor of Hsp90 dependent signal transduction, is anti-inflammatory in vivo implying that Hsp90 is critical for pathways involved in carrageenan-induced paw oedema. In addition the ability of GA to block NO release and reduce oedema formation suggests a therapeutic rationale for specific inhibitors of Hsp90 as potential anti-inflammatory drugs.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Quinonas/farmacología , Receptores de Glucocorticoides/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Animales , Benzoquinonas , Dexametasona/farmacología , Edema/prevención & control , Proteínas HSP90 de Choque Térmico/fisiología , Lactamas Macrocíclicas , Masculino , Ratones , Mifepristona/farmacologíaRESUMEN
1. Anti-inflammatory non steroidal drugs releasing NO (NO-NSAIDs) are a new class of anti-inflammatory drugs to which has been added an NO-releasing moiety. These compounds have been shown to retain the anti-inflammatory, analgesic and antipyretic activity of the parent compound but to be devoid of gastrointestinal (GI) toxicity. 2. Freund's adjuvant (FA) arthritis was induced in rats by a single intraplantar injection into the right hindpaw of 100 microl of mycobacterium butirricum (6 mg ml(-1)). The effect of equimolar doses of naproxen (1, 3 and 10 mg kg(-1)) and NO-naproxen (1.5, 4.5 and 16 mg kg(-1)) was evaluated using two dosage regimen protocols: (i) preventive, starting oral administration of the drugs at the time of induction of arthritis and for the following 21 days (day 1 - 21); (ii) therapeutic, starting oral administration of the drugs 7 days after adjuvant injection and for the following 14 days (day 7 - 21). 3. Hindpaw swelling (days 3, 7, 11, 14, 17, 21) and nociception (days 15 and 21) were measured. On day 22 rats were sacrificed, draining lymph nodes were removed and T cells isolated. In vitro proliferation of T cells following stimulation with concanavalin A (0.5 - 5 microg ml(-1)) was measured using a tritiated thymidine incorporation assay. IL-2 receptor expression on T cells was measured by FACS analysis. 4. Naproxen and NO-naproxen showed similar activity in reducing oedema formation in the non-injected (controlateral) hindpaw. Both drugs showed anti-nociceptive effect. NO-naproxen was anti-nociceptive at a dose of 4.5 mg kg(-1) while naproxen showed the same extent of inhibition only at a dose of 10 mg kg(-1). 5. T cells were isolated and characterized by FACS analysis. Stimulation of isolated T cells with concanavallin A in vitro caused a significant increase in thymidine uptake. NO-naproxen at a dose of 4.5 mg kg(-1) inhibited T cell proliferation to the same extent as 10 mg kg(-1) of naproxen. 6. Inhibition of T cell proliferation was well correlated with reduced IL-2 receptor expression on T cells. In addition, NO-naproxen reduced both IL-1beta and TNFalpha plasma levels whilst naproxen reduced IL-1beta levels only. 7. In conclusion, both naproxen and NO-naproxen reduce inflammation and nociception associated with arthritis. In addition NO-naproxen interferes to a larger extent with cellular mechanism involved in T cell activation in rat adjuvant arthritis indicating that introduction of the NO moiety in the naproxen structure increases the effect at the level of the immune system.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Artritis Experimental/prevención & control , Inflamación/prevención & control , Naproxeno/análogos & derivados , Óxido Nítrico/farmacología , Nociceptores/efectos de los fármacos , Linfocitos T/efectos de los fármacos , Animales , Artritis Experimental/inmunología , Artritis Experimental/fisiopatología , Relación Dosis-Respuesta a Droga , Edema/prevención & control , Citometría de Flujo , Miembro Posterior , Interleucina-1/sangre , Masculino , Naproxeno/farmacología , Dolor/prevención & control , Dimensión del Dolor , Ratas , Ratas Endogámicas Lew , Receptores de Interleucina-2/efectos de los fármacos , Receptores de Interleucina-2/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismo , Timidina/metabolismo , Factores de Tiempo , Factor de Necrosis Tumoral alfa/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The capacity of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) envelopes to transduce signals through chemokine coreceptors on macrophages was examined by measuring the ability of recombinant envelope proteins to mobilize intracellular calcium stores. Both HIV and SIV envelopes mobilized calcium via interactions with CCR5. The kinetics of these responses were similar to those observed when macrophages were treated with MIP-1beta. Distinct differences in the capacity of envelopes to mediate calcium mobilization were observed. Envelopes derived from viruses capable of replicating in macrophages mobilized relatively high levels of calcium, while envelopes derived from viruses incapable of replicating in macrophages mobilized relatively low levels of calcium. The failure to efficiently mobilize calcium was not restricted to envelopes derived from CXCR4-utilizing isolates but also included envelopes derived from CCR5-utilizing isolates that fail to replicate in macrophages. We characterized one CCR5-utilizing isolate, 92MW959, which entered macrophages but failed to replicate. A recombinant envelope derived from this virus mobilized low levels of calcium. When macrophages were inoculated with 92MW959 in the presence of MIP-1alpha, viral replication was observed, indicating that a CC chemokine-mediated signal provided the necessary stimulus to allow the virus to complete its replication cycle. Although the role that envelope-CCR5 signal transduction plays in viral replication is not yet understood, it has been suggested that envelope-mediated signals facilitate early postfusion events in viral replication. The data presented here are consistent with this hypothesis and suggest that the differential capacity of viral envelopes to signal through CCR5 may influence their ability to replicate in macrophages.
Asunto(s)
Productos del Gen env/fisiología , VIH/fisiología , Macrófagos/virología , Receptores CCR5/fisiología , Virus de la Inmunodeficiencia de los Simios/fisiología , Calcio/metabolismo , Células Cultivadas , Quimiocina CCL3 , Quimiocina CCL4 , Citometría de Flujo , Productos del Gen env/farmacología , VIH/patogenicidad , Humanos , Proteínas Inflamatorias de Macrófagos/farmacología , Macrófagos/fisiología , Monocitos/fisiología , Monocitos/virología , Receptores CCR5/metabolismo , Proteínas Recombinantes/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Replicación ViralRESUMEN
Following an injury, the body recruits a mechanism to delimit and repair tissue damage; this phenomenon is known as inflammation. Among the several different pathways that are activated during this process, which is necessary for survival, activation of the coagulation pathway is a key feature. In fact, clinical changes in blood fluidity have been closely related to ongoing inflammation. Recent evidence suggests that serine protease receptors might play a major role in the host defence mechanism at the interface between coagulation and inflammation.
Asunto(s)
Coagulación Sanguínea/fisiología , Inflamación/patología , Receptores de Trombina/metabolismo , Serina Endopeptidasas/metabolismo , Animales , Humanos , Receptor PAR-1 , Receptor PAR-2 , Receptores de Trombina/genética , Serina Endopeptidasas/químicaRESUMEN
Protease-activated receptor-2 (PAR-2) is a member of seven transmembrane domain G protein-coupled receptors activated by proteolytic cleavage whose better known member is the thrombin receptor. The pathophysiological role of PAR-2 remains poorly understood. Because PAR-2 is involved in inflammatory and injury response events, we investigated the role of PAR-2 in experimental myocardial ischemia-reperfusion injury. We show for the first time that PAR-2 activation protects against reperfusion-injury. After PAR-2-activating peptide (2AP) infusion, we found a significant recovery of myocardial function and decrease in oxidation at reflow. Indeed, the glutathione cycle (glutathione and oxidized glutathione) and lipid peroxidation analysis showed a reduced oxidative reperfusion-injury. Moreover, ischemic risk zone and creatine kinase release were decreased after PAR-2AP treatment. These events were coupled to elevation of PAR-2 and tumor necrosis factor alpha (TNFalpha) expression in both nuclear extracts and whole heart homogenates. The recovery of coronary flow was not reverted by L-nitroarginine methylester, indicating a NO-independent pathway for this effect. Genistein, a tyrosine kinase inhibitor, did not revert the PAR-2AP effect. During early reperfusion injury in vivo not only oxygen radicals are produced but also numerous proinflammatory mediators promoting neutrophil and monocyte targeting. In this context, we show that TNFalpha and PAR-2 are involved in signaling in pathophysiological conditions, such as myocardial ischemia-reperfusion. At the same time, because TNFalpha may exert pro-inflammatory actions and PAR-2 may constitute one of the first protective mechanisms that signals a primary inflammatory response, our data support the concept that this network may regulate body responses to tissue injury.
Asunto(s)
Isquemia Miocárdica/fisiopatología , Receptores de Trombina/fisiología , Daño por Reperfusión/fisiopatología , Animales , Western Blotting , Creatina Quinasa/metabolismo , Glutatión/metabolismo , Disulfuro de Glutatión/metabolismo , Masculino , Malondialdehído/metabolismo , Isquemia Miocárdica/enzimología , Isquemia Miocárdica/metabolismo , Ratas , Receptor PAR-2 , Receptores de Trombina/metabolismo , Daño por Reperfusión/enzimología , Daño por Reperfusión/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Rats injected in the hind paw with a mixture of Mycobacterium butirricum emulsified in mineral oil (FA) developed a severe polyarthritis that shared some immunological features with human rheumatoid arthritis. After this local administration, rats developed a secondary lesion (edema) in the contralateral paw, which is a hallmark of immune system activation. In vivo intravenous treatment with a monoclonal anti-very late antigen (VLA)-1 antibody (HA31/8) significantly reduced the edema formation in the contralateral paw. T cells isolated from contralateral paw draining lymph nodes of FA rats treated with HA31/8 showed a reduced cell proliferation in vitro, after stimulation with concanavalin A. Furthermore FACS analysis showed that the reduction in proliferation was concomitant to a reduction in the number of T cells positive to surface IL-2 receptor expression. Our data indicate that after in vivo treatment with a monoclonal anti-very late antigen-1 antibody, there is a beneficial effect on the development of the secondary lesion, which correlates to the reduced ability of T cells to proliferate in vitro as well as to a reduced surface expression of IL-2 receptor. The association of this antibody to other drugs interfering at other levels in rheumatoid arthritis may open a new therapeutic window.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Artritis Infecciosa/terapia , Integrinas/inmunología , Linfocitos T/inmunología , Animales , Artritis Infecciosa/sangre , Artritis Infecciosa/inmunología , Artritis Infecciosa/patología , Separación Celular , Citometría de Flujo , Humanos , Integrina alfa1beta1 , Interleucina-1/sangre , Masculino , Mycobacterium/patogenicidad , Ratas , Ratas Endogámicas Lew , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Binding of HIV type 1 (HIV-1) envelope glycoproteins to the surface of a CD4(+) T cell transduces intracellular signals through the primary envelope receptor, CD4, and a coreceptor, either CCR5 or CXCR4. Furthermore, envelope-CD4(+) cell interactions increase rates of apoptosis in peripheral blood mononuclear cells (PBMCs). We demonstrate that in primary T lymphocytes, recombinant HIV-1 envelope proteins induce the activation of caspase-3 and caspase-6, which belong to a family of cysteine proteases that, upon activation, promote programmed cell death. Envelope-mediated activation of caspase-3 and caspase-6 depended on envelope-CD4 receptor interactions; CCR5-utilizing as well as CXCR4-utilizing envelopes elicited this response. Focal adhesion kinase (FAK) is a substrate of both caspase-3 and caspase-6, and inactivation of FAK by these caspases promotes apoptosis. En-velope treatment of lymphocytes led to the cleavage of FAK in a manner consistent with caspase-mediated cleavage.
Asunto(s)
Linfocitos T CD4-Positivos/virología , Linfocitos T CD8-positivos/virología , Caspasas/metabolismo , Moléculas de Adhesión Celular/metabolismo , Productos del Gen env/fisiología , VIH-1/fisiología , Proteínas Tirosina Quinasas/metabolismo , Apoptosis , Linfocitos T CD4-Positivos/enzimología , Linfocitos T CD8-positivos/enzimología , Caspasa 3 , Caspasa 6 , Activación Enzimática , Quinasa 1 de Adhesión Focal , Proteína-Tirosina Quinasas de Adhesión Focal , Humanos , Fosforilación , Procesamiento Proteico-PostraduccionalRESUMEN
Initially described as an antiviral cytokine, IFN-alpha has been subsequently shown to affect several cellular functions, including cellular differentiation and proliferation. For these reasons, IFN-alpha is currently used in clinical practice for the treatment of viral infections and malignancies. In this manuscript, we show two novel mechanisms concomitantly responsible for the antiproliferative effect of IFN-alpha. First, long-term treatment with IFN-alpha of primary CD4+ T cells reduced surface expression of CD3 and CD28. These events resulted in decreased phosphorylation of the mitogen-activated extracellular signal-regulated activating kinase and its substrate extracellular signal-regulated kinase, leading to diminished production of IL-2. Second, IFN-alpha treatment of primary CD4+ T cells reduced proliferative response to stimulation in the presence of exogenous IL-2 by markedly decreasing mRNA synthesis and surface expression of CD25 (alpha-chain), a critical component of the IL-2R complex. These results may be relevant for the antitumor effects of IFN-alpha and may help us to better understand its detrimental role in the inhibition of proliferation of the bulk of CD4+ T cells (uninfected cells) in HIV-infected persons, who are known to overproduce IFN-alpha.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Interferón Tipo I/farmacología , Interleucina-2/antagonistas & inhibidores , Interleucina-2/biosíntesis , Receptores de Interleucina-2/antagonistas & inhibidores , Receptores de Interleucina-2/fisiología , Antígenos CD28/biosíntesis , Antígenos CD28/inmunología , Complejo CD3/biosíntesis , Complejo CD3/inmunología , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/enzimología , Membrana Celular/efectos de los fármacos , Membrana Celular/inmunología , Membrana Celular/metabolismo , Células Cultivadas , Humanos , Sueros Inmunes/farmacología , Interleucina-2/fisiología , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , MAP Quinasa Quinasa 1 , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosforilación/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/biosíntesis , Receptores de Interleucina-2/genética , Proteínas RecombinantesRESUMEN
The important role of chemokine receptors in HIV pathogenesis is becoming increasingly apparent. The level at which certain chemokine receptors that serve as HIV co-receptors are available influences the susceptibility of a CD4+ cell to viral infection and to certain HIV envelope-induced alterations in cellular function. Numerous pathogens, including HIV, can stimulate the production of chemokines and cytokines from a variety of cell types. Both cytokines and chemokines modulate CCR5 and CXCR4 availability, resulting in differential replication potentials for RS and X4 HIV strains depending on the milieu in the microenvironment. In addition, differential expression of CCR5 and CXCR4 on activated memory T cells appears to play an important role in preferential replication of RS HIV strains in vivo. However, expression of HIV co-receptors and CD4 may not be sufficient for effective HIV entry and replication. Intracellular signaling events, triggered by interaction between chemokine receptors and chemokines or HIV envelope, are important for efficient entry and completion of early replication events. Envelope proteins of different HIV isolates vary in their ability to transduce these signals, a characteristic that may play a role in determining the ability of a virus to productively infect certain cell types. Finally, the interaction between chemokine receptors and chemokines or HIV envelope has significant effects on cellular functions which likely play a role in HIV pathogenesis.
Asunto(s)
Citocinas/inmunología , Infecciones por VIH/inmunología , VIH-1/fisiología , VIH-2/fisiología , Humanos , Replicación Viral/inmunologíaRESUMEN
Binding of HIV-1 envelope glycoproteins to the surface of a CD4+ cell transduces intracellular signals through the primary envelope receptor, CD4, and/or the envelope coreceptor, a seven-transmembrane chemokine receptor. Macrophage-tropic strains of HIV-1 preferentially use CCR5 as an entry coreceptor, whereas T cell-tropic strains use CXC chemokine receptor-4 for entry. Intracellular signals transduced by HIV-1 envelope may have immunopathogenic consequences, including anergy, syncytium formation, apoptosis, and inappropriate cell trafficking. We demonstrate here that a recombinant envelope protein derived from an M-tropic isolate of HIV-1 can transduce CD4-dependent as well as CCR5-dependent intracellular signals in primary human CD4+ T cells. Novel HIV-induced intracellular signals that were identified include tyrosine phosphorylation of focal adhesion kinase (FAK) and CCR5, which are involved in cell adhesion and chemotaxis, respectively. HIV envelope-induced cellular association of FAK and CCR5 was also demonstrated, suggesting that ligation of CD4 and CCR5 leads to the formation of an activation complex composed of FAK and CCR5. Activation of this signaling pathway by HIV-1 envelope may be an important pathogenic mechanism of dysregulated cellular activation and trafficking during HIV infection.
Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Moléculas de Adhesión Celular/metabolismo , Productos del Gen env/fisiología , VIH/fisiología , Líquido Intracelular/metabolismo , Macrófagos/virología , Proteínas Tirosina Quinasas/metabolismo , Receptores CCR5/metabolismo , Antígenos CD4/inmunología , Antígenos CD4/metabolismo , Linfocitos T CD4-Positivos/enzimología , Linfocitos T CD4-Positivos/virología , Agregación Celular/inmunología , Línea Celular , Quinasa 1 de Adhesión Focal , Proteína-Tirosina Quinasas de Adhesión Focal , Humanos , Líquido Intracelular/inmunología , Ligandos , Fosfoproteínas/metabolismo , Fosforilación , Receptores CCR5/inmunología , Transducción de Señal/inmunología , Tirosina/metabolismoRESUMEN
BACKGROUND: The protease-activated receptor-2 (PAR-2) is expressed by vascular endothelial cells and upregulated by lipopolysaccharide (LPS) in vitro. PAR-2 is activated by a tethered ligand created after proteolytic cleavage by trypsin or experimentally by a synthetic agonist peptide (PAR-2AP) corresponding to the new amino terminus of the tethered ligand. METHODS AND RESULTS: Intravenous administration of PAR-2AP (0.1, 0.3, and 1 mg/kg) to rats caused a dose-dependent hypotension. A scrambled peptide was without effect. A specific trypsin inhibitor, biotin-SGKR-chloromethylketone, inhibited trypsin-induced hypotension but not that stimulated by PAR-2AP. In animals treated with LPS 20 hours earlier, we found an increased sensitivity to trypsin and PAR-2AP in the hypotensive response. In particular, PAR-2AP caused hypotension at a low concentration of 30 ng/kg. Moreover, PAR-2 was immunolocalized to endothelial and smooth muscle cells in aorta and jugular vein in LPS-treated rats, and increased levels of PAR-2 mRNA were shown by reverse transcription-polymerase chain reaction analysis. CONCLUSIONS: Our findings suggest that PAR-2 is important in the regulation of blood pressure in vivo. A functional upregulation of PAR-2 by LPS was demonstrated by the activity of concentrations of PAR-2AP that were inactive in normal animals. We conclude that PAR-2 may play an important role in the hypotension associated with endotoxic shock and may represent a new therapeutic target.
Asunto(s)
Endotoxemia/metabolismo , Hipotensión/metabolismo , Receptores de Trombina/metabolismo , Animales , Presión Sanguínea/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Endotoxemia/fisiopatología , Hipotensión/inducido químicamente , Hipotensión/fisiopatología , Lipopolisacáridos/administración & dosificación , Ratas , Receptor PAR-2 , Receptores de Trombina/administración & dosificaciónRESUMEN
1. Several thrombin cellular effects are dependent upon stimulation of proteinase activated receptor-1 (PAR-1) localized over the cellular surface. Following activation by thrombin, a new N-terminus peptide is unmasked on PAR-1 receptor, which functions as a tethered ligand for the receptor itself. Synthetic peptides called thrombin receptor activating peptides (TRAPs), corresponding to the N-terminus residue unmasked, reproduce several thrombin cellular effects, but are devoid of catalytic activity. We have evaluated the bronchial response to intravenous administration of human alpha-thrombin or a thrombin receptor activating peptide (TRAP-9) in anaesthetized, artificially ventilated guinea-pigs. 2. Intravenous injection of thrombin (100 microkg(-1)) caused bronchoconstriction that was recapitulated by injection of TRAP-9 (1 mg kg(-1)). Animal pretreatment with the thrombin inhibitor Hirulog (10 mg kg(-1) i.v.) prevented thrombin-induced bronchoconstriction, but did not affect bronchoconstriction induced by TRAP-9. Both agents did not induce bronchoconstriction when injected intravenously to rats. 3. The bronchoconstrictor effect of thrombin and TRAP-9 was subjected to tolerance; however, in animals desensitized to thrombin effect, TRAP-9 was still capable of inducing bronchoconstriction, but not vice versa. 4. Depleting animals of circulating platelets prevented bronchoconstriction induced by both thrombin and TRAP-9. 5. Bronchoconstriction was paralleled by a biphasic change in arterial blood pressure, characterized by a hypotensive phase followed by a hypertensive phase. Thrombin-induced hypotension was not subject to tolerance and was inhibited by Hirulog; conversely, hypertension was subject to tolerance and was not inhibited by Hirulog. Hypotension and hypertension induced by TRAP-9 were neither subject to tolerance nor inhibited by Hirulog. 6. Our results indicate that thrombin causes bronchoconstriction in guinea-pigs through a mechanism that requires proteolytic activation of its receptor and the exposure of the tethered ligand peptide. Platelet activation might be triggered by the thrombin effect.
Asunto(s)
Broncoconstricción/efectos de los fármacos , Broncoconstrictores/farmacología , Fragmentos de Péptidos/farmacología , Trombina/farmacología , Animales , Plaquetas/citología , Plaquetas/fisiología , Presión Sanguínea/efectos de los fármacos , Líquido del Lavado Bronquioalveolar/citología , Broncoconstricción/fisiología , Cobayas , Recuento de Leucocitos/efectos de los fármacos , Pulmón/efectos de los fármacos , Pulmón/patología , Recuento de Plaquetas , Receptores de Trombina/químicaRESUMEN
The effects of angiotensin-converting enzyme (ACE)-inhibition with zofenopril on the development of atherosclerosis and low-density lipoprotein (LDL) oxidation were determined in Watanabe Heritable Hyperlipidemic (WHHL) rabbits. Rabbits received either placebo (n = 6) or 0.5 mg/kg/day of zofenopril (n = 6). After 6 weeks of treatment, the computer-assisted analysis revealed that zofenopril reduced the aortic and common carotid corrected cumulative lesion area by 34% and 39%, respectively (p < 0.05 vs placebo-treated group). The intimal/medial ratio of the largest fatty streaks was 0.426+/-0.158 in the zofenopril-treated group and 0.875+/-0.238 in the placebo-treated group (p < 0.05). Furthermore, we found in the zofenopril-treated group smaller lesions with an intimal/medial ratio of zofenopril also reduced plasmatic LDL oxidation, as shown by significant reduction of malondialdehyde content (p < 0.01) and relative agarose gel mobility (p < 0.05), as well as by the prolongation of the lag-time (p < 0.05). Compared to zofenopril-treated rabbits, arterial sections of the placebo-group had significant increase in the intimal presence of macrophages-derived foam cells (p < 0.05), ox-LDL (p < 0.01), and native LDL (p < 0.01) detected by immunocytochemistry with RAM-11, MDA2 and NP1533975 monoclonal antibodies, respectively. To investigate the amount of platelet accumulation in the atherosclerotic plaque we also measured platelet-associated radioactivity. Autologous platelets were labeled with 111Indiumoxine and injected intravenously. After 2 hours, WHHL were sacrificed and arterial sections were counted for platelet-associated radioactivity. In the placebo-treated group, platelet radioactivity was 0.52+/-0.12 equivalent of radioactivity per mg of tissue in the common carotid and 0.25+/-0.18 in the abdominal aorta; in contrast, rabbits treated by zofenopril had 0.20+/-0.12 in the common carotid and 0.06+/-0.01 in the abdominal aorta. These data indicate that ACE-inhibition with zofenopril has antiatherosclerotic and antioxidant effects in WHHL-rabbits. Our results also shows that these effects could be linked to a reduced wall-associated platelet deposition at the site of atherosclerotic lesions.