RESUMEN
Sepsis concurrently activates both coagulation and complement systems. Although complement activation by bacteria is well documented, work in mice and in vitro suggests that coagulation proteases can directly cleave complement proteins. We aimed to determine whether generation of coagulation proteases in vivo can activate the complement cascade in 2 highly coagulopathic models. We compared temporal changes in activation biomarkers of coagulation (thrombin-antithrombin [TAT]), fibrinolysis (plasmin-antiplasmin [PAP]), and complement (C3b, C5a, C5b-9) in baboons infused with factor Xa (FXa) and phospholipids (FXa/phosphatidylcholine-phosphatidylserine [PCPS]) vs LD100 Escherichia coli We found that, albeit with different timing, both FXa/PCPS and E coli infusion led to robust thrombin and plasmin generation. Conversely, only E coli challenge activated the complement system, reaching a maximum at 2 hours postchallenge during the peaks of lipopolysaccharide and bacteremia but not of TAT and PAP. Despite inducing a strong burst of thrombin and plasmin, FXa/PCPS infusion did not produce measurable levels of complement activation in vivo. Similarly, ex vivo incubation of baboon serum with thrombin, plasmin, or FXa did not show noticeable complement cleavage unless supraphysiologic amounts of enzymes were used. Our results suggest that in vivo-generated thrombin and plasmin do not directly activate the complement in nonhuman primates.
Asunto(s)
Activación de Complemento/inmunología , Proteínas del Sistema Complemento/inmunología , Fibrinolisina/inmunología , Trombina/inmunología , Animales , Activación de Complemento/efectos de los fármacos , Proteínas del Sistema Complemento/metabolismo , Escherichia coli/inmunología , Escherichia coli/metabolismo , Factor Xa/inmunología , Factor Xa/farmacología , Fibrinolisina/metabolismo , Humanos , Lipopolisacáridos/inmunología , Lipopolisacáridos/farmacología , Papio , Fosfatidilcolinas/inmunología , Fosfatidilcolinas/farmacología , Fosfatidilserinas/inmunología , Fosfatidilserinas/farmacología , Trombina/metabolismoRESUMEN
Sphingosine 1-phosphate (S1P) is an important regulator of vascular integrity and immune cell migration, carried in plasma by high-density lipoprotein (HDL)-associated apolipoprotein M (apoM) and by albumin. In sepsis, the protein and lipid composition of HDL changes dramatically. The aim of this study was to evaluate changes in S1P and its carrier protein apoM during sepsis. For this purpose, plasma samples from both human sepsis patients and from an experimental Escherichia coli sepsis model in baboons were used. In the human sepsis cohort, previously studied for apoM, plasma demonstrated disease-severity correlated decreased S1P levels, the profile mimicking that of plasma apoM. In the baboons, a similar disease-severity dependent decrease in plasma levels of S1P and apoM was observed. In the lethal E. coli baboon sepsis, S1P decreased already within 6-8 hrs, whereas the apoM decrease was seen later at 12-24 hrs. Gel filtration chromatography of plasma from severe human or baboon sepsis on Superose 6 demonstrated an almost complete loss of S1P and apoM in the HDL fractions. S1P plasma concentrations correlated with the platelet count but not with erythrocytes or white blood cells. The liver mRNA levels of apoM and apoA1 decreased strongly upon sepsis induction and after 12 hr both were almost completely lost. In conclusion, during septic challenge, the plasma levels of S1P drop to very low levels. Moreover, the liver synthesis of apoM decreases severely and the plasma levels of apoM are reduced. Possibly, the decrease in S1P contributes to the decreased endothelial barrier function observed in sepsis.
Asunto(s)
Apolipoproteínas/metabolismo , Escherichia coli/fisiología , Lipocalinas/metabolismo , Lisofosfolípidos/metabolismo , Sepsis/metabolismo , Sepsis/microbiología , Esfingosina/análogos & derivados , Animales , Apolipoproteínas M , Plaquetas/metabolismo , Estudios de Casos y Controles , Cromatografía en Gel , Recuento de Colonia Microbiana , Eritrocitos/metabolismo , Humanos , Riñón/metabolismo , Leucocitos/metabolismo , Papio , ARN Mensajero/genética , ARN Mensajero/metabolismo , Sepsis/sangre , Esfingosina/metabolismo , Transcripción GenéticaRESUMEN
Acute respiratory distress syndrome (ARDS) induced by severe sepsis can trigger persistent inflammation and fibrosis. We have shown that experimental sepsis in baboons recapitulates ARDS progression in humans, including chronic inflammation and long-lasting fibrosis in the lung. Complement activation products may contribute to the fibroproliferative response, suggesting that complement inhibitors are potential therapeutic agents. We have been suggested that treatment of septic baboons with compstatin, a C3 convertase inhibitor protects against ARDS-induced fibroproliferation. Baboons challenged with 10(9) cfu/kg (LD50) live E. coli by intravenous infusion were treated or not with compstatin at the time of challenge or 5 hrs thereafter. Changes in the fibroproliferative response at 24 hrs post-challenge were analysed at both transcript and protein levels. Gene expression analysis showed that sepsis induced fibrotic responses in the lung as early as 24 hrs post-bacterial challenge. Immunochemical and biochemical analysis revealed enhanced collagen synthesis, induction of profibrotic factors and increased cell recruitment and proliferation. Specific inhibition of complement with compstatin down-regulated sepsis-induced fibrosis genes, including transforming growth factor-beta (TGF-ß), connective tissue growth factor (CTGF), tissue inhibitor of metalloproteinase 1 (TIMP1), various collagens and chemokines responsible for fibrocyte recruitment (e.g. chemokine (C-C motif) ligand 2 (CCL2) and 12 (CCL12)). Compstatin decreased the accumulation of myofibroblasts and proliferating cells, reduced the production of fibrosis mediators (TGF-ß, phospho-Smad-2 and CTGF) and inhibited collagen deposition. Our data demonstrate that complement inhibition effectively attenuates collagen deposition and fibrotic responses in the lung after severe sepsis. Inhibiting complement could prove an attractive strategy for preventing sepsis-induced fibrosis of the lung.
Asunto(s)
Bacteriemia/tratamiento farmacológico , Activación de Complemento/efectos de los fármacos , Inactivadores del Complemento/uso terapéutico , Infecciones por Escherichia coli/tratamiento farmacológico , Pulmón/patología , Péptidos Cíclicos/uso terapéutico , Animales , Bacteriemia/inmunología , Bacteriemia/patología , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/fisiopatología , Fibrosis , Regulación de la Expresión Génica/efectos de los fármacos , Síndrome de Dificultad Respiratoria/tratamiento farmacológico , Síndrome de Dificultad Respiratoria/inmunología , Síndrome de Dificultad Respiratoria/patologíaRESUMEN
We review our baboon models of Escherichia coli sepsis that mimic, respectively, the shock/disseminated intravascular coagulation (DIC) and organ failure variants of severe sepsis, and analyse the pathophysiologic processes that are unique to each. The multi-stage, multi-factorial characteristics of severe sepsis develop as a result of the initial insult, which - depending on its intensity - activates components of the intravascular compartment leading to overwhelming shock/DIC; or initiates a sequence of events involving both the intra- and extravascular (tissues) compartments that lead to organ failure. In the latter case, the disorder passes through two stages: an initial inflammatory/coagulopathic intravascular first stage triggered by E. coli, followed by an extravascular second stage, involving components unique to each organ and triggered by ischemia/reperfusion (oxidative stress and histone release). Although a myriad of overlapping cellular and molecular components are involved, it is the context in which these components are brought into play that determine whether shock/DIC or organ failure predominate. For example, inflammatory and thrombotic responses amplified by thrombin in the first case whereas similar responses are amplified by complement activation products in the second. Rather than blocking specific mediators, we found that attenuation of the thrombin and complement amplification pathways can effectively reverse the shock/DIC and organ failure exhibited by the LD(100) and LD(50) E. coli models of severe sepsis, respectively. Translation of these concepts to successful intervention in the respective baboon models of E. coli sepsis and the application to their clinical counterparts is described.
Asunto(s)
Modelos Animales de Enfermedad , Infecciones por Escherichia coli/fisiopatología , Infecciones por Escherichia coli/terapia , Sepsis/fisiopatología , Sepsis/terapia , Animales , Infecciones por Escherichia coli/microbiología , Estrés Oxidativo , Papio , Sepsis/microbiologíaRESUMEN
Severe sepsis leads to massive activation of coagulation and complement cascades that could contribute to multiple organ failure and death. To investigate the role of the complement and its crosstalk with the hemostatic system in the pathophysiology and therapeutics of sepsis, we have used a potent inhibitor (compstatin) administered early or late after Escherichia coli challenge in a baboon model of sepsis-induced multiple organ failure. Compstatin infusion inhibited sepsis-induced blood and tissue biomarkers of complement activation, reduced leucopenia and thrombocytopenia, and lowered the accumulation of macrophages and platelets in organs. Compstatin decreased the coagulopathic response by down-regulating tissue factor and PAI-1, diminished global blood coagulation markers (fibrinogen, fibrin-degradation products, APTT), and preserved the endothelial anticoagulant properties. Compstatin treatment also improved cardiac function and the biochemical markers of kidney and liver damage. Histologic analysis of vital organs collected from animals euthanized after 24 hours showed decreased microvascular thrombosis, improved vascular barrier function, and less leukocyte infiltration and cell death, all consistent with attenuated organ injury. We conclude that complement-coagulation interplay contributes to the progression of severe sepsis and blocking the harmful effects of complement activation products, especially during the organ failure stage of severe sepsis is a potentially important therapeutic strategy.
Asunto(s)
Coagulación Sanguínea/efectos de los fármacos , Proteínas Inactivadoras de Complemento/farmacología , Infecciones por Escherichia coli , Insuficiencia Multiorgánica/prevención & control , Péptidos Cíclicos/farmacología , Sepsis , Animales , Biomarcadores/sangre , Coagulación Sanguínea/inmunología , Presión Sanguínea/efectos de los fármacos , Activación de Complemento/efectos de los fármacos , Proteínas Inactivadoras de Complemento/metabolismo , Citocinas/sangre , Modelos Animales de Enfermedad , Infecciones por Escherichia coli/sangre , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/inmunología , Insuficiencia Multiorgánica/sangre , Insuficiencia Multiorgánica/inmunología , Papio , Sepsis/sangre , Sepsis/tratamiento farmacológico , Sepsis/inmunologíaRESUMEN
Activated thrombin-activatable fibrinolysis inhibitor (TAFIa or CPU) is a carboxypeptidase that is able to attenuate fibrinolysis. Although its role in fibrinolysis and inflammation has been studied extensively in vitro, its levels and subsequent effect in vivo has not been studied to the same extent. Using our recently developed assay that is specific for TAFIa, we were able to quantify its levels in plasma samples obtained from an Escherichia coli (E. coli) challenged baboon sepsis model. TAFIa levels accumulated appeared to be E. coli dose dependent, where the lethal dose of 10(10) CFU/kg generated a peak TAFIa level of 24 nM by 2 h, which represents almost 32% of total plasma level of its precursor, thrombin-activatable fibrinolysis inhibitor (TAFI or proCPU). Furthermore, our data suggest that there is continual TAFI activation under lethal level of E. coli as the apparent half-life of TAFIa is increased from 8 min to 2.2 h. Two sublethal doses of 10(8) and 10(6) CFU/kg generated peak TAFIa levels of 1.1 and 0.4 nM, respectively, both by 6 h. Taken together, our data show that TAFIa is generated at systemic levels, in a dose-dependent manner, that can substantially affect both fibrinolysis and inflammatory response in the E. coli challenged baboon sepsis model.
Asunto(s)
Carboxipeptidasa B2/sangre , Infecciones por Escherichia coli/sangre , Escherichia coli , Sepsis/sangre , Animales , Modelos Animales de Enfermedad , Activación Enzimática , Masculino , Papio cynocephalus , Sepsis/microbiología , Factores de TiempoRESUMEN
Twenty-two veterans with posttraumatic stress disorder (PTSD) were assessed for trauma-related nightmares and nonnightmare distressed awakenings (NNDA) before and after treatment with the alpha-1 adrenoreceptor antagonist prazosin at an average bedtime dose of 9.6 mg/day. Ratings combining frequency and intensity dimensions of trauma-related nightmares decreased from 3.6 to 2.2, NNDA from 5.2 to 2.1, and sleep difficulty from 7.2 to 4.1 per week. These results suggest that increased brain adrenergic activity may contribute to the pathophysiology of both trauma-related nightmares and NNDA in PTSD.
Asunto(s)
Antagonistas Adrenérgicos alfa/uso terapéutico , Sueños/efectos de los fármacos , Prazosina/uso terapéutico , Trastornos por Estrés Postraumático/fisiopatología , Veteranos/psicología , Antagonistas Adrenérgicos alfa/administración & dosificación , Humanos , Prazosina/administración & dosificación , Trastornos del Sueño-Vigilia/tratamiento farmacológicoRESUMEN
BACKGROUND: Bacterial invasion during sepsis induces disregulated systemic responses that could lead to fatal lung failure. The purpose of this study was to relate the temporal dynamics of gene expression to the pathophysiological changes in the lung during the first and second stages of E. coli sepsis in baboons. RESULTS: Using human oligonucleotide microarrays, we have explored the temporal changes of gene expression in the lung of baboons challenged with sublethal doses of E. coli. Temporal expression pattern and biological significance of the differentially expressed genes were explored using clustering and pathway analysis software. Expression of selected genes was validated by real-time PCR. Cytokine levels in tissue and plasma were assayed by multiplex ELISA. Changes in lung ultrastructure were visualized by electron microscopy. We found that genes involved in primary inflammation, innate immune response, and apoptosis peaked at 2 hrs. Inflammatory and immune response genes that function in the stimulation of monocytes, natural killer and T-cells, and in the modulation of cell adhesion peaked at 8 hrs, while genes involved in wound healing and functional recovery were upregulated at 24 hrs. CONCLUSION: The analysis of gene expression modulation in response to sepsis provides the baseline information that is crucial for the understanding of the pathophysiology of systemic inflammation and may facilitate the development of future approaches for sepsis therapy.
Asunto(s)
Análisis por Conglomerados , Infecciones por Escherichia coli/genética , Regulación de la Expresión Génica , Pulmón/metabolismo , Sepsis/genética , Biología de Sistemas , Animales , Apoptosis/genética , Adhesión Celular/genética , Modelos Animales de Enfermedad , Infecciones por Escherichia coli/inmunología , Humanos , Inmunidad/genética , Inflamación/genética , Pulmón/inmunología , Papio , Sepsis/inmunología , Factores de Tiempo , Cicatrización de Heridas/genéticaRESUMEN
BACKGROUND: Persons with posttraumatic stress disorder (PTSD) whose trauma-related nightmares improve or resolve with bedtime administration of the alpha-1 adrenergic antagonist prazosin often continue to experience PTSD symptoms during the day. This study addressed whether daytime prazosin compared to placebo would alleviate psychological distress provoked experimentally by a trauma-related word list included in the emotional Stroop (E-Stroop) paradigm. METHODS: Eleven persons with civilian trauma PTSD who continued to experience daytime PTSD symptoms despite a stable bedtime prazosin dose that suppressed trauma-related nightmares were studied. Prazosin and placebo were administered on two different occasions in the early afternoon followed two hours later by the E-Stroop. Effects of drug on psychological distress were assessed by the Profile of Mood States (POMS). RESULTS: POMS total score and an "emotional distress" POMS subscale score following trauma-related words were significantly lower in the prazosin than placebo condition. There were no treatment effects on E-Stroop completion time. In 10 subjects who continued open label daytime prazosin, there was a reduction in global PTSD illness severity at 2-week follow-up. CONCLUSIONS: Daytime prazosin pretreatment reduced psychological distress specifically to trauma cues. Adding daytime prazosin to bedtime prazosin may further reduce overall PTSD illness severity and distress.
Asunto(s)
Antagonistas Adrenérgicos alfa/uso terapéutico , Ritmo Circadiano , Señales (Psicología) , Depresión/tratamiento farmacológico , Depresión/etiología , Prazosina/uso terapéutico , Trastornos por Estrés Postraumático/psicología , Antagonistas de Receptores Adrenérgicos alfa 1 , Antagonistas Adrenérgicos alfa/farmacología , Depresión/diagnóstico , Manual Diagnóstico y Estadístico de los Trastornos Mentales , Método Doble Ciego , Sueños/psicología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Prazosina/farmacología , Índice de Severidad de la Enfermedad , Trastornos del Sueño-Vigilia/etiología , Trastornos por Estrés Postraumático/diagnóstico , Encuestas y CuestionariosRESUMEN
Research on the neurobiology of the stress response in animals has led to successful new treatments for Post-Traumatic Stress Disorder (PTSD) in humans. Basic research has found that high levels of catecholamine release during stress rapidly impair the top-down cognitive functions of the prefrontal cortex (PFC), while strengthening the emotional and habitual responses of the amygdala and basal ganglia. Chronic stress exposure leads to dendritic atrophy in PFC, dendritic extension in the amygdala, and strengthening of the noradrenergic (NE) system. High levels of NE release during stress engage low affinity alpha-1 adrenoceptors, (and likely beta-1 adrenoceptors), which rapidly reduce the firing of PFC neurons, but strengthen amygdala function. In contrast, moderate levels of NE release during nonstress conditions engage higher affinity alpha-2A receptors, which strengthen PFC, weaken amygdala, and regulate NE cell firing. Thus, either alpha-1 receptor blockade or alpha-2A receptor stimulation can protect PFC function during stress. Patients with PTSD have signs of PFC dysfunction. Clinical studies have found that blocking alpha-1 receptors with prazosin, or stimulating alpha-2A receptors with guanfacine or clonidine can be useful in reducing the symptoms of PTSD. Placebo-controlled trials have shown that prazosin is helpful in veterans, active duty soldiers and civilians with PTSD, including improvement of PFC symptoms such as impaired concentration and impulse control. Open label studies suggest that guanfacine may be especially helpful in treating children and adolescents who have experienced trauma. Thus, understanding the neurobiology of the stress response has begun to help patients with stress disorders.
RESUMEN
BACKGROUND: Inhibition of the tissue factor-factor VIIa pathway attenuated the activation of coagulation and prevented death in a gram-negative bacteremia primate model of sepsis. This lethal animal model suggested that tissue factor also influences inflammatory cascades. METHODS: This trial examined the pharmacodynamic effects of active site-inhibited factor VIIa (FFR-recombinant factor VIIa [rFVIIa]; ASIS) on endotoxin-induced procoagulant, fibrinolytic, and inflammatory responses in healthy humans. A double-blind, randomized, placebo-controlled, parallel-group study was conducted in 12 healthy male volunteers. Subjects received a bolus infusion of 2-ng/kg endotoxin, followed by a bolus infusion of ASIS (400 microg/kg) or placebo 10 minutes later. RESULTS: Endotoxin injection induced inflammation, activation of coagulation, and activation and subsequent inhibition of fibrinolysis. ASIS infusion completely blocked thrombin and fibrin generation, as measured by plasma levels of prothrombin fragment (no increase in the ASIS group, as compared with a 13-fold increase in the placebo group at 4 hours; P <.01), soluble fibrin, and fibrin split product D-dimer. ASIS did not alter endotoxin-induced changes in the fibrinolytic system, cytokine levels, or markers of endothelial (E-selectin, thrombomodulin) or platelet (P-selectin) activation. CONCLUSIONS: In summary, ASIS effectively and selectively attenuates tissue factor-induced thrombin generation. Because ASIS was well tolerated, this study provides seminal data to further characterize its anticoagulant and putative anti-inflammatory effects in critically ill patients.
Asunto(s)
Coagulación Sanguínea/efectos de los fármacos , Endotoxinas/farmacología , Factor VIIa/antagonistas & inhibidores , Adulto , Análisis de Varianza , Sitios de Unión/efectos de los fármacos , Sitios de Unión/fisiología , Coagulación Sanguínea/fisiología , Método Doble Ciego , Selectina E/sangre , Factor VIIa/administración & dosificación , Factor VIIa/efectos adversos , Fiebre/inducido químicamente , Fibrinolisina/antagonistas & inhibidores , Fibrinolisina/metabolismo , Humanos , Interleucina-6/sangre , Masculino , Selectina-P/sangre , Elastasa Pancreática/sangre , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/efectos adversos , Proteínas Recombinantes/antagonistas & inhibidores , Estadísticas no Paramétricas , Trombina/antagonistas & inhibidores , Trombina/metabolismo , Trombomodulina/sangre , Activador de Tejido Plasminógeno/sangre , Factor de Necrosis Tumoral alfa/metabolismo , alfa 2-Antiplasmina/metabolismoRESUMEN
BACKGROUND: Posttraumatic stress disorder (PTSD) is often a chronic disorder, and, though 2 antidepressants are now approved by the U.S. Food and Drug Administration for its treatment, it often remains refractory to pharmacotherapy. The memory of traumatic events, by repeatedly stimulating the hippocampus and amygdala (kindling phenomenon), may alter multiple biological systems, including gamma-aminobutyric acid (GABA) pathways, and eventually lead to the disorder. Tiagabine, a selective GABA reuptake inhibitor, was evaluated as a treatment for PTSD. METHOD: Patients with DSM-IV PTSD who were stable on current medications and still symptomatic were eligible for inclusion in this open-label case series. Tiagabine was initiated at 2 mg nightly and increased by 2-mg increments every 2 to 3 days until an optimal response was achieved. The Clinical Global Impressions-Improvement scale and PTSD Checklist-Civilian Version (PCL-C) were used to evaluate changes in PTSD symptoms. RESULTS: Seven consecutive female patients were identified as eligible. Tiagabine markedly improved PTSD symptoms within 2 weeks for 6 of the 7 patients, and 6 patients were rated as "much improved" or "very much improved." The mean PCL-C score was significantly reduced at weeks 2 and 8 (p <.05) as were the 3 PCL-C subscales and 1 of 2 items related to sleep disturbance. The mean effective daily dosage was approximately 8 mg (range, 4-12 mg/day). Treatment with tiagabine was generally well tolerated. CONCLUSIONS: These preliminary open-label findings suggest that the selective GABA reuptake inhibitor tiagabine may be a promising therapeutic option in the treatment of PTSD. Further study into the efficacy and safety of tiagabine for the treatment of PTSD is warranted.
Asunto(s)
Agonistas del GABA/uso terapéutico , Ácidos Nipecóticos/uso terapéutico , Trastornos por Estrés Postraumático/tratamiento farmacológico , Adulto , Antidepresivos/administración & dosificación , Antidepresivos/uso terapéutico , Comorbilidad , Trastorno Depresivo Mayor/diagnóstico , Trastorno Depresivo Mayor/tratamiento farmacológico , Trastorno Depresivo Mayor/psicología , Quimioterapia Combinada , Femenino , Agonistas del GABA/efectos adversos , Humanos , Persona de Mediana Edad , Ácidos Nipecóticos/efectos adversos , Trastornos por Estrés Postraumático/diagnóstico , Trastornos por Estrés Postraumático/psicología , Tiagabina , Resultado del TratamientoRESUMEN
To monitor and better understand the immunoinflammatory sequelae in sepsis and septic shock, systemic and monocyte-related cytokine responses were evaluated in baboons with experimental peritonitis induced by an E. coli-laden fibrin clot. Despite similar bacterial inocula, considerable interindividual variability in clinical manifestation and outcome of infection was observed. Because monocytes and macrophages are a key component of innate immunity, we hypothesized that early polarization of distinct activation programs in circulating monocytes that culminates in the emergence of either classically (M1) or alternatively (M2) activated monocytes may underlie the observed susceptibility or resistance to infection. To test our hypothesis, we analyzed infection-induced expression of cytokine mRNAs in monocytes isolated from surviving and dead animals. Our data show that resistance to E. coli sepsis may well be associated with a mixed M1/M2 activation state of circulating monocytes, whereas M1 phenotype appeared to be prevailing in monocytes from animals that died. Together with data on systemic cytokine responses, the latter findings indicate that morbidity and mortality of animals with gram-negative sepsis may well result from an overwhelming proinflammatory response. Collectively, our data contribute to a better understanding of cytokine networking in the immunoinflammatory response to microbial infection and suggest M1/M2 immunophenotypic profiling of readily available circulatory monocytes for early prognosis of severe infections.
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Monocitos/inmunología , Monocitos/microbiología , Sepsis/sangre , Sepsis/diagnóstico , Animales , Línea Celular , Células Cultivadas , Citocinas/metabolismo , Cartilla de ADN/química , Endotoxinas/metabolismo , Escherichia coli/metabolismo , Fibrina/química , Inflamación , Leucocitos Mononucleares/citología , Ratones , Monocitos/citología , Monocitos/metabolismo , Papio , Fenotipo , Reacción en Cadena de la Polimerasa , Pronóstico , ARN/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sepsis/patología , Factores de TiempoAsunto(s)
Proteínas del Sistema Complemento/inmunología , Endotoxemia/inmunología , Infecciones por Escherichia coli/inmunología , Hemostasis/inmunología , Factores Inmunológicos/inmunología , Inflamación/inmunología , Sepsis/inmunología , Animales , Citocinas/sangre , Citocinas/inmunología , Endotoxemia/sangre , Infecciones por Escherichia coli/sangre , Humanos , Inflamación/sangre , Pulmón/inmunología , Pulmón/patología , Papio , Sepsis/sangreRESUMEN
Hyperinflammatory responses can lead to a variety of diseases, including sepsis. We now report that extracellular histones released in response to inflammatory challenge contribute to endothelial dysfunction, organ failure and death during sepsis. They can be targeted pharmacologically by antibody to histone or by activated protein C (APC). Antibody to histone reduced the mortality of mice in lipopolysaccharide (LPS), tumor necrosis factor (TNF) or cecal ligation and puncture models of sepsis. Extracellular histones are cytotoxic toward endothelium in vitro and are lethal in mice. In vivo, histone administration resulted in neutrophil margination, vacuolated endothelium, intra-alveolar hemorrhage and macro- and microvascular thrombosis. We detected histone in the circulation of baboons challenged with Escherichia coli, and the increase in histone levels was accompanied by the onset of renal dysfunction. APC cleaves histones and reduces their cytotoxicity. Co-infusion of APC with E. coli in baboons or histones in mice prevented lethality. Blockade of protein C activation exacerbated sublethal LPS challenge into lethality, which was reversed by treatment with antibody to histone. We conclude that extracellular histones are potential molecular targets for therapeutics for sepsis and other inflammatory diseases.
Asunto(s)
Líquido Extracelular/metabolismo , Histonas/metabolismo , Sepsis/mortalidad , Sepsis/patología , Animales , Anticuerpos/farmacología , Anticuerpos/uso terapéutico , Bovinos , Línea Celular Transformada , Modelos Animales de Enfermedad , Endotelio/efectos de los fármacos , Endotelio/patología , Endotelio/ultraestructura , Escherichia coli/fisiología , Líquido Extracelular/efectos de los fármacos , Citometría de Flujo , Hemorragia/etiología , Hemorragia/patología , Histonas/efectos de los fármacos , Histonas/inmunología , Histonas/farmacología , Enfermedades Renales/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Macrófagos/ultraestructura , Ratones , Microscopía Electrónica de Transmisión/métodos , Neutrófilos/efectos de los fármacos , Neutrófilos/patología , Oligopéptidos/farmacología , Oligopéptidos/uso terapéutico , Papio , Polisacáridos/efectos adversos , Sepsis/tratamiento farmacológico , Sepsis/etiología , Factor de Necrosis Tumoral alfa/efectos adversosRESUMEN
BACKGROUND: Prazosin, a central nervous system (CNS) active alpha-1 adrenoreceptor antagonist, has reduced nightmares and sleep disturbance in placebo-controlled studies of combat-related posttraumatic stress disorder (PTSD). We evaluated objective sleep parameters and PTSD symptoms in a placebo-controlled prazosin trial for civilian trauma-related PTSD. METHODS: Thirteen outpatients with chronic civilian trauma PTSD, frequent nightmares, and sleep disturbance participated in a randomized placebo-controlled crossover trial of prazosin. Sleep parameters were quantified at home with the REMView (Respironics, Pittsburgh, Pennsylvania). The PTSD symptoms were quantified with the Clinician Administered PTSD Scale (CAPS) "recurrent distressing dreams" and "disturbed sleep" items, a non-nightmare distressed awakenings scale, the PTSD Dream Rating Scale (PDRS), the PTSD Checklist-Civilian (PCL-C), and the Clinical Global Impression of Improvement (CGI-I). RESULTS: Prazosin compared with placebo significantly increased total sleep time by 94 min; increased rapid eye movement (REM) sleep time and mean REM period duration without altering sleep onset latency; significantly reduced trauma-related nightmares, distressed awakenings, and total PCL scores; significantly improved CGI-I scores; and changed PDRS scores toward normal dreaming. CONCLUSIONS: Prazosin reductions of nighttime PTSD symptoms in civilian trauma PTSD are accompanied by increased total sleep time, REM sleep time, and mean REM period duration in the absence of a sedative-like effect on sleep onset latency.
Asunto(s)
Antagonistas Adrenérgicos alfa/uso terapéutico , Prazosina/uso terapéutico , Trastornos del Inicio y del Mantenimiento del Sueño/tratamiento farmacológico , Sueño/efectos de los fármacos , Trastornos por Estrés Postraumático/diagnóstico , Antagonistas Adrenérgicos alfa/efectos adversos , Adulto , Estudios Cruzados , Sueños/efectos de los fármacos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Polisomnografía , Prazosina/efectos adversos , Trastornos del Inicio y del Mantenimiento del Sueño/diagnóstico , Trastornos del Inicio y del Mantenimiento del Sueño/psicología , Sueño REM/efectos de los fármacos , Trastornos por Estrés Postraumático/psicología , Vigilia/efectos de los fármacosRESUMEN
Thrombin activatable fibrinolysis inhibitor (TAFI), when activated, forms a basic carboxypeptidase that can inhibit fibrinolysis. Potential physiologic activators include both thrombin and plasmin. In vitro, thrombomodulin and glycosaminoglycans increase the catalytic efficiency of TAFI activation by thrombin and plasmin, respectively. The most relevant (patho-) physiologic activator of TAFI has not been disclosed. Our purpose was to identify the physiologic activator of TAFI in vivo. Activation of protein C (a thrombin-thrombomodulin-dependent reaction), prothrombin, and plasminogen occurs during sepsis. Thus, a baboon model of Escherichia coli-induced sepsis, where multiple potential activators of TAFI are elaborated, was used to study TAFI activation. A monoclonal antibody (mAbTAFI/TM#16) specifically inhibiting thrombin-thrombomodulin-dependent activation of TAFI was used to assess the contribution of thrombin-thrombomodulin in TAFI activation in vivo. Coinfusion of mAbTAFI/TM#16 with a lethal dose of E coli prevented the complete consumption of TAFI observed without mAbTAFI/TM#16. The rate of fibrin degradation products formation is enhanced in septic baboons treated with the mAbTAFI/TM#16; therefore, TAFI activation appears to play a key role in the extent of fibrin(ogen) consumption during E coli challenge, and thrombin-thrombomodulin, in a baboon model of E coli-induced sepsis, appears to be the predominant activator of TAFI.
Asunto(s)
Coagulación Sanguínea/fisiología , Carboxipeptidasa B2/metabolismo , Fibrinólisis/fisiología , Trombina/fisiología , Trombomodulina/fisiología , Animales , Anticuerpos Monoclonales/farmacología , Carboxipeptidasa B2/antagonistas & inhibidores , Carboxipeptidasa B2/sangre , Carboxipeptidasa B2/inmunología , Infecciones por Escherichia coli/sangre , Infecciones por Escherichia coli/patología , Productos de Degradación de Fibrina-Fibrinógeno/metabolismo , Fibrinógeno/metabolismo , Semivida , Humanos , Pruebas de Sensibilidad Microbiana , Papio cynocephalus , Sepsis/sangre , Sepsis/patologíaRESUMEN
Increased tissue factor (TF)-dependent procoagulant activity in sepsis may be partly due to decreased expression or function of tissue factor pathway inhibitor (TFPI). To test this hypothesis, baboons were infused with live Escherichia coli and sacrificed after 2, 8, or 24 hours. Confocal and electron microscopy revealed increased leukocyte infiltration and fibrin deposition in the intravascular and interstitial compartments. Large amounts of TF were detected by immunostaining in leukocytes and platelet-rich microthrombi. TF induction was documented by quantitative reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent assay, and coagulation assays. Lung-associated TFPI antigen and mRNA decreased during sepsis, and TFPI activity diminished abruptly at 2 hours. Blocking antibodies against TFPI increased fibrin deposition in septic baboon lungs, suggesting that TF-dependent coagulation might be aggravated by reduced endothelial TFPI. Decreased TFPI activity coincided with the release of tissue plasminogen activator and the peak of plasmin generation, suggesting that TFPI could undergo proteolytic inactivation by plasmin. Enhanced plasmin produced in septic baboons by infusion of blocking antibodies against plasminogen activator inhibitor-1 led to decreased lung-associated TFPI and unforeseen massive fibrin deposition. We conclude that activation of TF-driven coagulation not adequately countered by TFPI may underlie the widespread thrombotic complications of sepsis.
Asunto(s)
Anticoagulantes/metabolismo , Coagulación Sanguínea , Lipoproteínas/metabolismo , Pulmón/metabolismo , Pulmón/patología , Papio cynocephalus , Sepsis , Animales , Anticuerpos/metabolismo , Escherichia coli/inmunología , Fibrinolisina/metabolismo , Humanos , Pulmón/citología , Pulmón/microbiología , Macrófagos/citología , Macrófagos/metabolismo , Neutrófilos/citología , Neutrófilos/metabolismo , Inhibidor 1 de Activador Plasminogénico/metabolismoRESUMEN
PRIMARY OBJECTIVE: Antisense oligomers to NF-kappaB (ASO) were incorporated into albumin microspheres to determine if microcapsules containing ASO inhibit pro-inflammatory cytokines to a greater extent than comparable doses of ASO in solution. Phagocytosis of microcapsules and intracellular release of ASO in macrophages was evaluated. RESEARCH DESIGN: Comparable doses of microencapsulated ASO and ASO in solution were evaluated in non-human primates. METHODS: Blood was sampled and stimulated with Escherichia coli endotoxin ex vivo. TNF, IL-1 and IL-6 concentrations were compared for 72 hrs. The intracellular concentration of ASO was measured in macrophages in vitro to evaluate the difference in intracellular penetration of microencapsulated ASO. RESULTS: Microencapsulated ASO produced significantly greater cytokine inhibition at all time points compared to ASO in solution. There were no side effects to ASO in the baboons. Intracellular ASO concentration was 10 fold greater in macrophages using microencapsulation. CONCLUSIONS: Microencapsulated ASO to NF-kappaB is more effective than ASO in solution in pro-inflammatory cytokine inhibition in non-human primates.
Asunto(s)
Cápsulas , Citocinas/antagonistas & inhibidores , Inflamación/fisiopatología , FN-kappa B/genética , Oligonucleótidos Antisentido/farmacología , Análisis de Varianza , Animales , Emulsiones , Macrófagos/efectos de los fármacos , Macrófagos/fisiología , Ratones , Microesferas , Papio , Albúmina Sérica BovinaRESUMEN
Studies that define natural responses to bacterial sepsis assumed new relevance after the lethal bioterrorist attacks with Bacillus anthracis (anthrax), a spore-forming, toxigenic gram-positive bacillus. Considerable effort has focused on identifying adjunctive therapeutics and vaccines to prevent future deaths, but translation of promising compounds into the clinical setting necessitates an animal model that recapitulates responses observed in humans. Here we describe a nonhuman primate (Papio c. cynocephalus) model of B. anthracis infection using infusion of toxigenic B. anthracis Sterne 34F2 bacteria (5 x 10(5) to 6.5 x 10(9) CFU/kg). Similar to that seen in human patients, we observed changes in vascular permeability, disseminated intravascular coagulation, and systemic inflammation. The lung was a primary target organ with serosanguinous pleural effusions, intra-alveolar edema, and hemorrhagic lesions. This animal model reveals that a fatal outcome is dominated by the host septic response, thereby providing important insights into approaches for treatment and prevention of anthrax in humans.