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Immunocompromised populations are highly vulnerable to developing life-threatening infections. Strategies to protect patients with weak immune responses are urgently needed. Employing trained immunity, whereby innate leukocytes undergo reprogramming upon exposure to a microbial product and respond more robustly to subsequent infection, is a promising approach. Previously, we demonstrated that the TLR4 agonist monophosphoryl lipid A (MPLA) induces trained immunity and confers broad resistance to infection. TLR4 signals through both MyD88- and TRIF-dependent cascades, but the relative contribution of each pathway to induction of trained immunity is unknown. Here, we show that MPLA-induced resistance to Staphylococcus aureus infection is lost in MyD88-KO, but not TRIF-KO, mice. The MyD88-activating agonist CpG (TLR9 agonist), but not TRIF-activating Poly I:C (TLR3 agonist), protects against infection in a macrophage-dependent manner. MPLA- and CpG-induced augmentation of macrophage metabolism and antimicrobial functions is blunted in MyD88-, but not TRIF-KO, macrophages. Augmentation of antimicrobial functions occurs in parallel to metabolic reprogramming and is dependent, in part, on mTOR activation. Splenic macrophages from CpG-treated mice confirmed that TLR/MyD88-induced reprogramming occurs in vivo. TLR/MyD88-triggered metabolic and functional reprogramming was reproduced in human monocyte-derived macrophages. These data show that MyD88-dependent signaling is critical in TLR-mediated trained immunity.
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Factor 88 de Diferenciación Mieloide , Receptor Toll-Like 4 , Humanos , Ratones , Animales , Factor 88 de Diferenciación Mieloide/metabolismo , Receptor Toll-Like 4/metabolismo , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Receptores Toll-Like/metabolismo , Macrófagos , Proteínas Adaptadoras Transductoras de Señales/metabolismoRESUMEN
Methyl CpG binding protein 2 (MeCP2) is involved in nerve regeneration following ischemic stroke, but the related mechanism remains unclear. Here, we found low MeCP2 expression in hippocampal tissues. Using functional analysis, we demonstrated that MeCP2 accelerated FOXO3a methylation and subsequently inhibited its expression, thus repressing the apoptosis of neuronal cells. Mechanistically, FOXO3a could bind to the promoter region of SPRY2, consequently inducing its transcription and promoting the expression of the downstream target gene ZEB1. Altogether, our study revealed that overexpression of MeCP2 can protect mice against ischemic brain injury via disruption of the FOXO3a/SPRY2/ZEB1 signaling axis. Our results identify ectopic expression of MeCP2 as a therapeutic target in ischemic stroke.
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Proteína Forkhead Box O3/metabolismo , Accidente Cerebrovascular Isquémico , Proteína 2 de Unión a Metil-CpG , Animales , Metilación de ADN , Proteína 2 de Unión a Metil-CpG/genética , Proteína 2 de Unión a Metil-CpG/metabolismo , Metilación , Ratones , Regiones Promotoras Genéticas , Transducción de SeñalRESUMEN
Bacterial infections are a common and deadly threat to vulnerable patients. Alternative strategies to fight infection are needed. ß-Glucan, an immunomodulator derived from the fungal cell wall, provokes resistance to infection by inducing trained immunity, a phenomenon that persists for weeks to months. Given the durability of trained immunity, it is unclear which leukocyte populations sustain this effect. Macrophages have a life span that surpasses the duration of trained immunity. Thus, we sought to define the contribution of differentiated macrophages to trained immunity. Our results show that ß-glucan protects mice from Pseudomonas aeruginosa infection by augmenting recruitment of innate leukocytes to the site of infection and facilitating local clearance of bacteria, an effect that persists for more than 7 d. Adoptive transfer of macrophages, trained using ß-glucan, into naive mice conferred a comparable level of protection. Trained mouse bone marrow-derived macrophages assumed an antimicrobial phenotype characterized by enhanced phagocytosis and reactive oxygen species production in parallel with sustained enhancements in glycolytic and oxidative metabolism, increased mitochondrial mass, and membrane potential. ß-Glucan induced broad transcriptomic changes in macrophages consistent with early activation of the inflammatory response, followed by sustained alterations in transcripts associated with metabolism, cellular differentiation, and antimicrobial function. Trained macrophages constitutively secreted CCL chemokines and robustly produced proinflammatory cytokines and chemokines in response to LPS challenge. Induction of the trained phenotype was independent of the classic ß-glucan receptors Dectin-1 and TLR-2. These findings provide evidence that ß-glucan induces enhanced protection from infection by driving trained immunity in macrophages.
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Memoria Inmunológica/efectos de los fármacos , Macrófagos/efectos de los fármacos , Sustancias Protectoras/farmacología , beta-Glucanos/farmacología , Animales , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/inmunología , Femenino , Inmunidad Innata/efectos de los fármacos , Inmunidad Innata/inmunología , Memoria Inmunológica/inmunología , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
BACKGROUND: Peripherally inserted central catheters (PICCs) have been increasingly applied worldwide owing to many advantages. Even with these advantages, the related complications should not be ignored, especially in neonates. The available evidence about PICC-related thrombosis was manifold, but the cardiac tamponade, an emergency and life-threatening complication, has been rarely reported. Early recognized cardiac tamponade by ultrasound may reduce mortality. CASE SUMMARY: A neonate weighting 2.8 kg was born at 40 wk of gestation. He was admitted to the Surgery Intensive Care Unit due to suspected congenital megacolon. A PICC line was inserted via the left antecubital fossa for the administration of total parenteral nutrition. Three days later, the patient was still on total parenteral nutrition. Cardiac tamponade caused by PICC was found on ultrasound. The patient recovered spontaneously after an emergency pericardiocentesis. CONCLUSION: Proficiency in the use of point-of-care ultrasound may save the life of patients, since it enables clinicians to treat patients faster, more accurately, and in a non-invasive way at the point of care.
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An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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Critically ill, severely injured and high-risk surgical patients are vulnerable to secondary infections during hospitalization and after hospital discharge. Studies show that the mitochondrial function and oxidative metabolism of monocytes and macrophages are impaired during sepsis. Alternatively, treatment with microbe-derived ligands, such as monophosphoryl lipid A (MPLA), peptidoglycan, or ß-glucan, that interact with toll-like receptors and other pattern recognition receptors on leukocytes induces a state of innate immune memory that confers broad-spectrum resistance to infection with common hospital-acquired pathogens. Priming of macrophages with MPLA, CPG oligodeoxynucleotides (CpG ODN), or ß-glucan induces a macrophage metabolic phenotype characterized by mitochondrial biogenesis and increased oxidative metabolism in parallel with increased glycolysis, cell size and granularity, augmented phagocytosis, heightened respiratory burst functions, and more effective killing of microbes. The mitochondrion is a bioenergetic organelle that not only contributes to energy supply, biosynthesis, and cellular redox functions but serves as a platform for regulating innate immunological functions such as production of reactive oxygen species (ROS) and regulatory intermediates. This review will define current knowledge of leukocyte metabolic dysfunction during and after sepsis and trauma. We will further discuss therapeutic strategies that target leukocyte mitochondrial function and might have value in preventing or reversing sepsis- and trauma-induced immune dysfunction.
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Infecciones/inmunología , Leucocitos/metabolismo , Mitocondrias/metabolismo , Sepsis/inmunología , Heridas y Lesiones/inmunología , Animales , Reprogramación Celular , Humanos , Inmunidad Innata , Leucocitos/inmunología , Estrés OxidativoRESUMEN
Ischemic cerebrovascular disease is a significant and common public health issue worldwide. The emerging roles of mesenchymal stem cells (MSCs)-derived extracellular vesicles (EVs) in ischemic neuronal injury continue to be investigated. The current study aimed to investigate the role of EV-derived miR-132 from MSCs in ischemic neuronal injury. EVs were initially isolated from bone MSCs (BMSCs) and subsequently evaluated. A middle cerebral artery occlusion (MCAO) mouse model was constructed with the neurological function evaluated through a series of neurological scores, a pole test, and a foot fault test. Histopathological changes, neuron viability, and apoptosis, as well as cerebral infarction, were detected by hematoxylin and eosin (HE) staining and 2,3,5-triphenyltetrazolium hydrochloride (TTC) staining. The targeting relationship between microRNA (miR)-132 and Activin receptor type IIB (Acvr2b) was further confirmed based on dual-luciferase reporter gene assay results. Loss- and gain-of-function assays were conducted to elucidate the role of miR-132, EV-derived miR-132, Acvr2b, and Smad2 in oxygen-glucose deprivation (OGD)-treated neurons, and in mice models. Neuronal cell viability and apoptosis were evaluated via Cell Counting kit-8 (CCK-8) and flow cytometry. Our results indicated that Acvr2b was highly expressed, while miR-132 was poorly expressed in the MCAO mice and OGD-treated neurons. Acvr2b silencing or upregulation of miR-132 led to an elevation in neuronal activity, decreased neuronal apoptosis, reduced expression of Bax, and cleaved-caspase 3, as well as increased Bcl-2 expression. Acvr2b expression was targeted and inhibited by miR-132. EV-derived Acvr2b promoted activation of phosphorylated-Smad2 (p-Smad2)/c-jun signaling pathway, ultimately inducing neuronal injury. Our study provides evidence demonstrating that the overexpression of c-jun inhibits the protective role of MSCs-derived EV-miR-132 in neuronal injury. Upregulation of EV-derived miR-132 released from MSCs attenuates ischemic neuronal injury by inhibiting Smad2/c-jun pathways via the suppression of Acvr2b.
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STUDY OBJECTIVE: Postoperative delirium (POD) is a common neurological system disorder in surgical patients. Anesthesia providers have a wide choice of sedative agents involving different mechanisms in clinical practice, and the incidence of POD varies regarding which sedative agent administered. This network meta-analysis aimed to comprehensively analyze the safety and efficacy of each choice for patients. DESIGN: A network meta-analysis. SETTING: Vanderbilt University Medical Center. MEASUREMENTS: We searched PubMed, EMBASE, Ovid Medline and Cochrane Central Register of Controlled Trials (CENTRAL) through the end of September 2018 with the registration number CRD42018110585. The randomized controlled trials were identified and extracted by two reviewers independently. Commonly used sedative agents such as placebo, sevoflurane, desflurane, isoflurane, dexmedetomidine, propofol, midazolam, and ketamine were assessed in this network meta-analysis and the primary outcome was the incidence of POD. The data were synthesized by network meta-analysis. Pair-wise meta-analyses were conducted using the random-effects model. Each intervention was ranked according to its corresponding surface under the cumulative ranking curve (SUCRA) values. The GRADE framework was undertaken to evaluate the risk of bias. MAIN RESULTS: We identified 39 RCTs and 5991 patients in this meta-analysis. Dexmedetomidine was found to be the most effective option in reducing POD, compared to midazolam, propofol, desflurane, and sevoflurane. The results revealed that dexmedetomidine was associated with a lower incidence of POD, whereas midazolam was associated with a significantly higher number of patients with delirium. Midazolam and propofol were also associated with a higher incidence of perioperative hypotension and bradycardia. CONCLUSION: Our study provided meta-analytic evidence and suggested dexmedetomidine could be considered as the most effective sedative agent to reduce POD. However, clinical practitioners still need to weigh the pros and cons before choosing a sedative agent for individual patient.
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Anestesia General/efectos adversos , Anestésicos Generales/efectos adversos , Delirio del Despertar/epidemiología , Hipnóticos y Sedantes/administración & dosificación , Atención Perioperativa/efectos adversos , Anestesia General/métodos , Anestésicos Generales/administración & dosificación , Dexmedetomidina/administración & dosificación , Dexmedetomidina/efectos adversos , Delirio del Despertar/etiología , Delirio del Despertar/prevención & control , Humanos , Hipnóticos y Sedantes/efectos adversos , Incidencia , Metaanálisis en Red , Atención Perioperativa/métodos , Náusea y Vómito Posoperatorios/inducido químicamente , Náusea y Vómito Posoperatorios/epidemiologíaRESUMEN
OBJECTIVES: Cardiopulmonary bypass-induced endothelial dysfunction has been inferred by changes in pulmonary vascular resistance, alterations in circulating biomarkers, and postoperative capillary leak. Endothelial-dependent vasomotor dysfunction of the systemic vasculature has never been quantified in this setting. The objective of the present study was to quantify acute effects of cardiopulmonary bypass on endothelial vasomotor control and attempt to correlate these effects with postoperative cytokines, tissue edema, and clinical outcomes in infants. DESIGN: Single-center prospective observational cohort pilot study. SETTING: Pediatric cardiac ICU at a tertiary children's hospital. PATIENTS: Children less than 1 year old requiring cardiopulmonary bypass for repair of a congenital heart lesion. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: Laser Doppler perfusion monitoring was coupled with local iontophoresis of acetylcholine (endothelium-dependent vasodilator) or sodium nitroprusside (endothelium-independent vasodilator) to quantify endothelial-dependent vasomotor function in the cutaneous microcirculation. Measurements were obtained preoperatively, 2-4 hours, and 24 hours after separation from cardiopulmonary bypass. Fifteen patients completed all laser Doppler perfusion monitor (Perimed, Järfälla, Sweden) measurements. Comparing prebypass with 2-4 hours postbypass responses, there was a decrease in both peak perfusion (p = 0.0006) and area under the dose-response curve (p = 0.005) following acetylcholine, but no change in responses to sodium nitroprusside. Twenty-four hours after bypass responsiveness to acetylcholine improved, but typically remained depressed from baseline. Conserved endothelial function was associated with higher urine output during the first 48 postoperative hours (R = 0.43; p = 0.008). CONCLUSIONS: Cutaneous endothelial dysfunction is present in infants immediately following cardiopulmonary bypass and recovers significantly in some patients within 24 hours postoperatively. Confirmation of an association between persistent endothelial-dependent vasomotor dysfunction and decreased urine output could have important clinical implications. Ongoing research will explore the pattern of endothelial-dependent vasomotor dysfunction after cardiopulmonary bypass and its relationship with biochemical markers of inflammation and clinical outcomes.
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Puente Cardiopulmonar/efectos adversos , Enfermedades Cardiovasculares/etiología , Endotelio Vascular/fisiopatología , Sistema Vasomotor/fisiopatología , Acetilcolina/uso terapéutico , Biomarcadores/sangre , Procedimientos Quirúrgicos Cardíacos/efectos adversos , Enfermedades Cardiovasculares/tratamiento farmacológico , Niño , Preescolar , Citocinas/sangre , Endotelio Vascular/metabolismo , Cardiopatías Congénitas/cirugía , Humanos , Lactante , Microcirculación , Óxido Nítrico/sangre , Proyectos Piloto , Complicaciones Posoperatorias/etiología , Estudios Prospectivos , Índice de Severidad de la Enfermedad , Resistencia Vascular , Vasodilatadores/uso terapéutico , Sistema Vasomotor/metabolismoRESUMEN
OBJECTIVE: To test the hypothesis that patient-controlled analgesia (PCA) contributes to improvement of hemorheology in patients undergoing hip arthroplasty. METHODS: 120 patients, aged 60â-â75 years old, undergoing hip arthroplasty under spinal anesthesia, were randomly divided into group PCA (n = 60) and control group (n = 60). Patients in PCA group received PCA in postoperative 3 days. Blood samples from the median cubital vein were collected at five time points: before anesthesia (T1), after surgery (T2), 6 h after surgery (T3), 24 h after surgery (T4), 48 h after surgery (T5). Hemorheological parameters were measured, including whole blood viscosity at a high shear rate (Hηb), whole blood viscosity at a low shear rate (Lηb), reduced viscosity (ηr), plasma viscosity (ηp), hematocrit (Hct), erythrocyte aggregation index(EAI) and erythrocyte deformation index (EDI). Noninvasive blood pressure and heart rate at T1-5 and pain scoring of visual analogue scale (VAS) score at T2-5 were recorded. RESULTS: (1) Compared with T1, Hηb, Lηb, ηp, ηr decreased significantly at T3-5 with EAI decreased significantly at T5 in group PCA (p < 0.05), EDI increased significantly at T5 in group C (p < 0.05). (2) Compared with group C, Hηb, Lηb, ηp, ηr, EAI decreased significantly at T5 with Lηb concurrently decreased at T4 in group PCA (p < 0.05). CONCLUSION: Postoperative pain may increase blood viscosity in patients undergoing hip arthroplasty, mainly via plasma viscosity, erythrocyte aggregation and rigidity, and which could be improved by postoperative PCA.
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Artroplastia de Reemplazo de Cadera , Anciano , Analgesia Controlada por el Paciente , Hemorreología , Humanos , Persona de Mediana Edad , Dolor PostoperatorioRESUMEN
BACKGROUND: Monophosphoryl lipid A (MPLA) is a TLR4 agonist that has potent immunomodulatory properties and modulates innate immune function to improve host resistance to infection with common nosocomial pathogens in mice. The goal of this study was to assess the safety and efficacy of MPLA in a sheep model of burn injury and Pseudomonas aeruginosa pneumonia. The sheep provides a favorable model for preclinical testing as their response to TLR4 agonists closely mimics that of humans. METHODS: Twelve chronically instrumented adult female Merino sheep received 20% total body surface area, third-degree cutaneous burn under anesthesia and analgesia. At 24âh after burn, sheep were randomly allocated to receive: MPLA (2.5âµg/kg i.v., nâ=â6), or vehicle (i.v., nâ=â6). At 24âh after MPLA or vehicle treatment, Pseudomonas aeruginosa pneumonia was induced. Sheep were mechanically ventilated, fluid resuscitated and cardiopulmonary variables were monitored for 24âh after induction of pneumonia. Cytokine production, vascular barrier function, and lung bacterial burden were also measured. RESULTS: MPLA infusion induced small and transient alterations in core body temperature, heart rate, pulmonary artery pressure, and pulmonary vascular resistance. Pulmonary mechanics were not altered. Vehicle-treated sheep developed severe acute lung injury during Pseudomonas aeruginosa pneumonia, which was attenuated by MPLA as indicated by improved PaO2/FiO2 ratio, oxygenation index, and shunt fraction. Sheep treated with MPLA also exhibited less vascular leak, lower blood lactate levels, and lower modified organ injury score. MPLA treatment attenuated systemic cytokine production and decreased lung bacterial burden. CONCLUSIONS: MPLA was well tolerated in burned sheep and attenuated development of acute lung injury, lactatemia, cytokinemia, vascular leak, and hemodynamic changes caused by Pseudomonas aeruginosa pneumonia.
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Adyuvantes Inmunológicos/uso terapéutico , Quemaduras/complicaciones , Lípido A/análogos & derivados , Insuficiencia Multiorgánica/prevención & control , Neumonía Bacteriana/complicaciones , Infecciones por Pseudomonas/complicaciones , Animales , Modelos Animales de Enfermedad , Femenino , Lípido A/uso terapéutico , Insuficiencia Multiorgánica/etiología , Neumonía Bacteriana/microbiología , Pseudomonas aeruginosa , OvinosRESUMEN
Infectious diseases remain a threat to critically ill patients, particularly with the rise of antibiotic-resistant bacteria. Septic shock carries a mortality of up to â¼40% with no compelling evidence of promising therapy to reduce morbidity or mortality. Septic shock survivors are also prone to nosocomial infections. Treatment with toll-like receptor 4 (TLR4) agonists have demonstrated significant protection against common nosocomial pathogens in various clinically relevant models of infection and septic shock. TLR4 agonists are derived from a bacteria cell wall or synthesized de novo, and more recently novel small molecule TLR4 agonists have also been developed. TLR4 agonists augment innate immune functions including expansion and recruitment of innate leukocytes to the site of infection. Recent studies demonstrate TLR4-induced leukocyte metabolic reprogramming of cellular metabolism to improve antimicrobial function. Metabolic changes include sustained augmentation of macrophage glycolysis, mitochondrial function, and tricarboxylic acid cycle flux. These findings set the stage for the use of TLR4 agonists as standalone therapeutic agents or antimicrobial adjuncts in patient populations vulnerable to nosocomial infections.
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Adyuvantes Inmunológicos/uso terapéutico , Resistencia a la Enfermedad/inmunología , Receptor Toll-Like 4/agonistas , Animales , Humanos , Inmunidad Innata , Control de Infecciones , Infecciones/inmunología , Receptor Toll-Like 4/inmunologíaRESUMEN
OBJECTIVES: To determine whether synthetic phosphorylated hexa-acyl disaccharides provide antimicrobial protection in clinically relevant models of bacterial infection. DESIGN: Laboratory study. SETTING: University laboratory. SUBJECTS: BALB/c, C57BL/10J, and C57BL/10ScNJ mice. INTERVENTIONS: Mice were treated with lactated Ringer's (vehicle) solution, monophosphoryl lipid A, or phosphorylated hexa-acyl disaccharides at 48 and 24 hours prior to intraperitoneal Pseudomonas aeruginosa or IV Staphylococcus aureus infection. Leukocyte recruitment, cytokine production, and bacterial clearance were measured 6 hours after P. aeruginosa infection. In the systemic S. aureus infection model, one group of mice was monitored for 14-day survival and another for S. aureus tissue burden at 3 days postinfection. Duration of action for 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide was determined at 3, 10, and 14 days using a model of intraperitoneal P. aeruginosa infection. Effect of 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide on in vivo leukocyte phagocytosis and respiratory burst was examined. Leukocyte recruitment, cytokine production, and bacterial clearance were measured after P. aeruginosa infection in wild-type and toll-like receptor 4 knockout mice treated with 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide or vehicle to assess receptor specificity. MEASUREMENTS AND MAIN RESULTS: During intraperitoneal P. aeruginosa infection, phosphorylated hexa-acyl disaccharides significantly attenuated infection-induced hypothermia, augmented leukocyte recruitment and bacterial clearance, and decreased cytokine production. At 3 days post S. aureus infection, bacterial burden in lungs, spleen, and kidneys was significantly decreased in mice treated with monophosphoryl lipid A or phosphorylated hexa-acyl disaccharides, which was associated with improved survival. Leukocyte phagocytosis and respiratory burst functions were enhanced after treatment with monophosphoryl lipid A or phosphorylated hexa-acyl disaccharides. A time course study showed that monophosphoryl lipid A- and 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide-mediated protection against P. aeruginosa lasts for up to 10 days. Partial loss of augmented innate antimicrobial responses was observed in toll-like receptor 4 knockout mice treated with 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide. CONCLUSIONS: Phosphorylated hexa-acyl disaccharides significantly augment resistance against clinically relevant Gram-negative and Gram-positive infections via enhanced leukocyte recruitment, phagocytosis, and respiratory burst functions of innate leukocytes. Improved antimicrobial protection persists for up to 10 days and is partially mediated through toll-like receptor 4.
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Infección Hospitalaria/prevención & control , Citocinas/metabolismo , Disacáridos/farmacología , Hexosaminidasa A/farmacología , Cavidad Peritoneal/fisiopatología , Infecciones Estafilocócicas/fisiopatología , Análisis de Varianza , Animales , Western Blotting/métodos , Modelos Animales de Enfermedad , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Cavidad Peritoneal/microbiología , Distribución Aleatoria , Infecciones Estafilocócicas/mortalidad , Estadísticas no Paramétricas , Tasa de SupervivenciaRESUMEN
Monophosphoryl lipid A (MPLA) is a clinically used TLR4 agonist that has been found to drive nonspecific resistance to infection for up to 2 wk. However, the molecular mechanisms conferring protection are not well understood. In this study, we found that MPLA prompts resistance to infection, in part, by inducing a sustained and dynamic metabolic program in macrophages that supports improved pathogen clearance. Mice treated with MPLA had enhanced resistance to infection with Staphylococcus aureus and Candida albicans that was associated with augmented microbial clearance and organ protection. Tissue macrophages, which exhibited augmented phagocytosis and respiratory burst after MPLA treatment, were required for the beneficial effects of MPLA. Further analysis of the macrophage phenotype revealed that early TLR4-driven aerobic glycolysis was later coupled with mitochondrial biogenesis, enhanced malate shuttling, and increased mitochondrial ATP production. This metabolic program was initiated by overlapping and redundant contributions of MyD88- and TRIF-dependent signaling pathways as well as downstream mTOR activation. Blockade of mTOR signaling inhibited the development of the metabolic and functional macrophage phenotype and ablated MPLA-induced resistance to infection in vivo. Our findings reveal that MPLA drives macrophage metabolic reprogramming that evolves over a period of days to support a macrophage phenotype highly effective at mediating microbe clearance and that this results in nonspecific resistance to infection.
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Macrófagos/metabolismo , Receptor Toll-Like 4/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Candida albicans/efectos de los fármacos , Candidiasis/tratamiento farmacológico , Candidiasis/metabolismo , Glucólisis/fisiología , Lípido A/análogos & derivados , Macrófagos/microbiología , Masculino , Ratones , Ratones Endogámicos C57BL , Factor 88 de Diferenciación Mieloide/metabolismo , Transducción de Señal/fisiología , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/metabolismo , Staphylococcus aureus/efectos de los fármacos , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
Burn patients are susceptible to infections due, in part, to immune dysfunction. Upregulation of programmed death-1 (PD-1) receptor on T cells and programmed cell death ligand-1 (PD-L1) on myeloid cells contribute to immune dysfunction in nonburn-related sepsis. We hypothesized that PD-1/PDL1 interactions contribute to immune dysfunction after burn injury. To determine the impact of burn injury and infection on PD-L1, PD-1 and costimulatory receptor expression by leukocytes and its relationship to T cell functions. The efficacy of anti-PD-L1 antibody was evaluated in a clinically relevant mouse model of burn injury and bacterial infection. Mice underwent 35% scald burn followed by Pseudomonas aeruginosa or Staphylococcus aureus infection on day 4 postburn. Anti-PD-L1 was administered on day 3 postburn. Numbers and phenotype of leukocytes, plasma cytokine concentrations, bacterial clearance, organ injury, and survival were assessed. Burn injury and infection with P. aeruginosa caused a significant upregulation of PD-L1 on myeloid cells, along with a decrease in T cell numbers and function, significant multiorgan injury, and decreased survival. Treatment with anti-PD-L1 antibody improved bacterial clearance, reduced organ injury, and enhanced survival during Pseudomonas burn wound infection. Furthermore, anti-PD-L1 effectively protected against multiorgan injury, and improved bacterial clearance and survival following systemic S. aureus infection after burn injury. Blockade of PD-1/PD-L1 interactions might represent a viable treatment to improve outcomes among critically ill burn-injured subjects and increased leukocyte PD-L1 expression could serve as a valuable biomarker to select appropriate patients for such treatment.
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Anticuerpos Monoclonales/farmacología , Antígeno B7-H1/antagonistas & inhibidores , Quemaduras/complicaciones , Infecciones por Pseudomonas/tratamiento farmacológico , Infecciones Estafilocócicas/tratamiento farmacológico , Linfocitos T/inmunología , Infección de Heridas/tratamiento farmacológico , Animales , Antígeno B7-H1/inmunología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Masculino , Ratones , Ratones Endogámicos BALB C , Infecciones por Pseudomonas/etiología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/aislamiento & purificación , Infecciones Estafilocócicas/etiología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/aislamiento & purificación , Linfocitos T/efectos de los fármacos , Linfocitos T/metabolismo , Infección de Heridas/etiologíaRESUMEN
Natural killer (NK) cells are large granular lymphocytes largely recognized for their importance in tumour surveillance and the host response to viral infections. However, as the major innate lymphocyte population, NK cells also coordinate early responses to bacterial infections by amplifying the antimicrobial functions of myeloid cells, especially macrophages, by production of interferon-γ (IFN-γ). Alternatively, excessive NK cell activation and IFN-γ production can amplify the systemic inflammatory response during sepsis resulting in increased physiological dysfunction and organ injury. Our understanding of NK cell biology during bacterial infections and sepsis is mostly derived from studies performed in mice. Human studies have demonstrated a correlation between altered NK cell functions and outcomes during sepsis. However, mechanistic understanding of NK cell function during human sepsis is limited. In this review, we will review the current understanding of NK cell biology during sepsis and discuss the challenges associated with modulating NK cell function during sepsis for therapeutic benefit.
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Infecciones Bacterianas/inmunología , Interferón gamma/inmunología , Células Asesinas Naturales/inmunología , Activación de Linfocitos , Sepsis/inmunología , Animales , Infecciones Bacterianas/patología , Infecciones Bacterianas/terapia , Humanos , Células Asesinas Naturales/patología , Ratones , Sepsis/patología , Sepsis/terapiaRESUMEN
Immunosuppression is increasingly being recognized as one of the causes of increased morbidity and mortality during sepsis. Both innate and adaptive immune system dysfunction have been shown to cause an impaired ability to eradicate the primary infection and also lead to frequent occurrence of secondary opportunistic infections. Pre-clinical and clinical studies have shown that inhibitory immune checkpoint molecules, including programmed death-1 (PD-1), programmed death ligand-1 (PD-L1), cytotoxic T lymphocyte antigen-4 (CTLA-4), T cell membrane protein-3 (TIM-3), Lymphocyte activation-gene-3 (LAG-3) and 2B4, are upregulated during the course of sepsis. Engagement of these inhibitory molecules on various immune cells has been consistently shown to inhibit innate immune cell functions (e.g., phagocytosis, cytokine production and pathogen clearance) and also lead to impaired T cell competence. In numerous pre-clinical models of sepsis, therapeutic agents aimed at blocking engagement of inhibitory immune checkpoints on immune cells have been shown to improve innate and adaptive immune cell functions, increase host resistance to infection and significantly improve survival. Therefore, immunotherapy with immune cell checkpoint inhibitors holds significant potential for the future of sepsis therapy and merits further investigation.
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Puntos de Control del Ciclo Celular , Sepsis/inmunología , Sepsis/patología , Animales , Biomarcadores/metabolismo , Humanos , Modelos InmunológicosRESUMEN
MicroRNA-mediated post-transcriptional regulation plays key roles in stem cell self-renewal and tumorigenesis. However, the in vivo functions of specific microRNAs in controlling mammary stem cell (MaSC) activity and breast cancer formation remain poorly understood. Here we show that miR-31 is highly expressed in MaSC-enriched mammary basal cell population and in mammary tumors, and is regulated by NF-κB signaling. We demonstrate that miR-31 promotes mammary epithelial proliferation and MaSC expansion at the expense of differentiation in vivo. Loss of miR-31 compromises mammary tumor growth, reduces the number of cancer stem cells, as well as decreases tumor-initiating ability and metastasis to the lung, supporting its pro-oncogenic function. MiR-31 modulates multiple signaling pathways, including Prlr/Stat5, TGFß and Wnt/ß-catenin. Particularly, it activates Wnt/ß-catenin signaling by directly targeting Wnt antagonists, including Dkk1. Importantly, Dkk1 overexpression partially rescues miR31-induced mammary defects. Together, these findings identify miR-31 as the key regulator of MaSC activity and breast tumorigenesis.
Asunto(s)
Neoplasias de la Mama/metabolismo , MicroARNs/metabolismo , Células Madre Neoplásicas/citología , Células Madre/metabolismo , Proteínas Wnt/metabolismo , Animales , Neoplasias de la Mama/genética , Neoplasias de la Mama/fisiopatología , Línea Celular Tumoral , Proliferación Celular , Autorrenovación de las Células , Regulación hacia Abajo , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Glándulas Mamarias Humanas/citología , Glándulas Mamarias Humanas/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , MicroARNs/genética , FN-kappa B/genética , FN-kappa B/metabolismo , Células Madre Neoplásicas/metabolismo , Células Madre/citología , Proteínas Wnt/genética , Vía de Señalización Wnt , beta Catenina/metabolismoRESUMEN
Interleukin (IL)-15 is essential for natural killer (NK), NKT and memory (m) CD8+ T cell development and function, and is currently under investigation as an immunotherapeutic agent for the treatment of cancer. Recently, the creation of IL-15 superagonist by complexing IL-15 and its high affinity receptor alpha (IL-15 Rα) in solution, inspired by the natural trans-presentation of IL-15, advances the potential of IL-15-based tumor immunotherapy. IL-15 superagonist shows promising advantages over monomeric IL-15 such as sustaining high circulating concentrations due to prolonged half-life and more potently stimulating NK and CD8+ T effector lymphocytes. So far, there are three different forms of recombinant IL-15 superagonist fusion protein based on configurational modifications. Gene therapy using engineered cells co-expressing IL-15/IL-15 Rα complex for cancer treatment is also emerging. All forms have demonstrated efficacy in causing tumor regression in animal studies, which provides strong rationale for advancing IL-15 superagonist through clinical trials. To date, there are fourteen phase I/II IL-15 superagonist trials in cancer patients and one phase I trial in HIV patients. Information generated by ongoing trials regarding the toxicity and efficacy of IL-15 superagonist is awaited. Finally, we elaborate on immunotoxicity caused by IL-15 superagonist in preclinical studies and discuss important safety considerations.
Asunto(s)
Subunidad alfa del Receptor de Interleucina-15/inmunología , Interleucina-15/inmunología , Neoplasias/inmunología , Virosis/inmunología , Animales , Terapia Genética , Humanos , Interleucina-15/agonistas , Interleucina-15/genética , Subunidad alfa del Receptor de Interleucina-15/genética , Neoplasias/tratamiento farmacológico , Virosis/tratamiento farmacológicoRESUMEN
Emerging evidence indicates that even subtle changes in the expression of key genes of signalling pathways can have profound effects. MicroRNAs (miRNAs) are masters of subtlety and generally have only mild effects on their target genes. The microRNA miR-31 is one of the major microRNAs in many cutaneous conditions associated with activated keratinocytes, such as the hyperproliferative diseases psoriasis, non-melanoma skin cancer and hair follicle growth. miR-31 is a marker of the hair growth phase, and in our miR-31 transgenic mouse model it impairs the function of keratinocytes. This leads to aberrant proliferation, apoptosis, and differentiation that results in altered hair growth, while the loss of miR-31 leads to increased hair growth. Through in vitro and in vivo studies, we have defined a set of conserved miR-31 target genes, including LATS2 and STK40, which serve as new players in the regulation of keratinocyte growth and hair follicle biology. LATS2 can regulate growth of keratinocytes and we have identified a function of STK40 that can promote the expression of key hair follicle programme regulators such as HR, DLX3 and HOXC13.