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1.
J Infect Dis ; 227(12): 1364-1375, 2023 06 15.
Artículo en Inglés | MEDLINE | ID: mdl-36763010

RESUMEN

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection triggers activation of the NLRP3 inflammasome, which promotes inflammation and aggravates severe COVID-19. Here, we report that SARS-CoV-2 induces upregulation and activation of human caspase-4/CASP4 (mouse caspase-11/CASP11), and this process contributes to NLRP3 activation. In vivo infections performed in transgenic hACE2 humanized mice, deficient or sufficient for Casp11, indicate that hACE2 Casp11-/- mice were protected from disease development, with the increased pulmonary parenchymal area, reduced clinical score of the disease, and reduced mortality. Assessing human samples from fatal cases of COVID-19, we found that CASP4 was expressed in patient lungs and correlated with the expression of inflammasome components and inflammatory mediators, including CASP1, IL1B, IL18, and IL6. Collectively, our data establish that CASP4/11 promotes NLRP3 activation and disease pathology, revealing a possible target for therapeutic interventions for COVID-19.


Asunto(s)
COVID-19 , Inflamasomas , Ratones , Animales , Humanos , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Macrófagos/metabolismo , COVID-19/metabolismo , SARS-CoV-2/metabolismo , Ratones Transgénicos
2.
Cytokine ; 142: 155494, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-33765652

RESUMEN

Interleukin-6 (IL-6) is associated with pathological cardiac hypertrophy and can be dramatically increased in serum after an acute strenuous exercise session. However, IL-6 is also associated with the increased production and release of anti-inflammatory cytokines and the inhibition of tumor necrosis factor-alpha (TNF-α) after chronic moderate exercise. To elucidate the relevance of IL-6 in inflammatory and hypertrophic signaling in the heart in response to an acute strenuous exercise session, we combined transcriptome analysis using the BXD mice database and exercised IL-6 knockout mice (IL-6KO). Bioinformatic analysis demonstrated that low or high-levels of Il6 mRNA in the heart did not change the inflammation- and hypertrophy-related genes in BXD mice strains. On the other hand, bioinformatic analysis revealed a strong positive correlation between Il6 gene expression in skeletal muscle with inflammation-related genes in cardiac tissue in several BXD mouse strains, suggesting that skeletal muscle-derived IL-6 could alter the heart's intracellular signals, particularly the inflammatory signaling. As expected, an acute strenuous exercise session increased IL-6 levels in wild-type, but not in IL-6KO mice. Despite not showing morphofunctional differences in the heart at rest, the IL-6KO group presented a reduction in physical performance and attenuated IL-6, TNF-α, and IL-1beta kinetics in serum, as well as lower p38MAPK phosphorylation, Ampkalpha expression, and higher Acta1 and Tnf gene expressions in the left ventricle in the basal condition. In response to strenuous exercise, IL-6 ablation was linked to a reduction in the pro-inflammatory response and higher activation of classical physiological cardiac hypertrophy proteins.


Asunto(s)
Biomarcadores/metabolismo , Corazón/fisiopatología , Inflamación/patología , Interleucina-6/deficiencia , Condicionamiento Físico Animal , Adenilato Quinasa/metabolismo , Animales , Biomarcadores/sangre , Cardiomegalia/sangre , Cardiomegalia/genética , Electrocardiografía , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Corazón/diagnóstico por imagen , Interleucina-6/genética , Interleucina-6/metabolismo , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Esquelético/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Descanso , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo
3.
Inflamm Res ; 69(1): 105-113, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31754736

RESUMEN

OBJECTIVE AND DESIGN: Investigate survival outcomes, and immunological and metabolomic effects of hyaluronidase (Hz) treatment during mouse models of acute inflammation and sepsis. METHODS: Survival of C57Bl/6 mice was monitored after lethal challenge with lipopolysaccharide (LPS) or cecal and ligation puncture (CLP)-induced sepsis and treated with Hz or saline. Mice were also challenged with LPS and treated with Hz for leukocyte counting, cytokine quantification and determination of metabolomic profiles in the peritoneal fluid. RESULTS: Hz treatment improved survival outcomes after lethal challenge with LPS or CLP-induced sepsis. LPS challenge promoted acute neutrophil accumulation and production of interleukin-1ß (IL-1ß) and IL-6 in the peritoneum, whereas Hz treatment suppressed neutrophil infiltration and cytokine production. We further characterized the metabolomic alterations caused by LPS challenge, which predicted activity of metabolic pathways related to fatty acids and eicosanoids. Hz treatment had a profound effect over the metabolic response, reflected by reductions of the relative levels of fatty acids. CONCLUSION: Collectively, these data demonstrate that Hz treatment is associated with metabolic reprogramming of pathways that sustain the inflammatory response.


Asunto(s)
Hialuronoglucosaminidasa/farmacología , Sepsis/inmunología , Sepsis/metabolismo , Enfermedad Aguda , Animales , Líquido Ascítico/citología , Líquido Ascítico/inmunología , Líquido Ascítico/metabolismo , Modelos Animales de Enfermedad , Eicosanoides/metabolismo , Ácidos Grasos/metabolismo , Inmunomodulación , Interleucina-1beta/inmunología , Interleucina-6/inmunología , Recuento de Leucocitos , Lipopolisacáridos , Masculino , Redes y Vías Metabólicas/efectos de los fármacos , Metabolómica , Ratones Endogámicos C57BL
4.
Cytokine ; 88: 99-107, 2016 12.
Artículo en Inglés | MEDLINE | ID: mdl-27591510

RESUMEN

Although much research has been done related to biomarker discovery for tuberculosis infection, a set of biomarkers that can discriminate between active and latent TB diseases remains elusive. In the current study we correlate clinical aspects of TB disease with changes in the immune response as determined by biomarkers detected in plasma. Our study measured 18 molecules in human plasma in 17 patients with active disease (APTB), 14 individuals with latent tuberculosis infection (LTBI) and 16 uninfected controls (CTRL). We found that active tuberculosis patients have increased plasma levels of IL-6, IP-10, TNF-α, sCD163 and sCD14. Statistical analysis of these biomarkers indicated that simultaneous measurement of sCD14 and IL-6 was able to diagnose active tuberculosis infection with 83% accuracy. We also demonstrated that TNF-α and sCD163 were correlated with tuberculosis severity. We showed that the simultaneous detection of both plasma sCD14 and IL-6 is a promising diagnostic approach to identify APTB, and further, measurement of TNF-α and sCD163 can identify the most severe cases of tuberculosis.


Asunto(s)
Citocinas/sangre , Receptores de Lipopolisacáridos/sangre , Tetraspanina 30/sangre , Tuberculosis Pulmonar/sangre , Adulto , Biomarcadores/sangre , Femenino , Humanos , Masculino
5.
Arch Toxicol ; 90(5): 1261-78, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-26026608

RESUMEN

Coagulopathies following snakebite are triggered by pro-coagulant venom toxins, in which metalloproteases play a major role in envenomation-induced coagulation disorders by acting on coagulation cascade, platelet function and fibrinolysis. Considering this relevance, here we describe the isolation and biochemical characterization of moojenactivase (MooA), a metalloprotease from Bothrops moojeni snake venom, and investigate its involvement in hemostasis in vitro. MooA is a glycoprotein of 85,746.22 Da, member of the PIIId group of snake venom metalloproteases, composed of three linked disulfide-bonded chains: an N-glycosylated heavy chain, and two light chains. The venom protease induced human plasma clotting in vitro by activating on both blood coagulation factors II (prothrombin) and X, which in turn generated α-thrombin and factor Xa, respectively. Additionally, MooA induced expression of tissue factor (TF) on the membrane surface of peripheral blood mononuclear cells (PBMC), which led these cells to adopt pro-coagulant characteristics. MooA was also shown to be involved with production of the inflammatory mediators TNF-α, IL-8 and MCP-1, suggesting an association between MooA pro-inflammatory stimulation of PBMC and TF up-regulation. We also observed aggregation of washed platelets when in presence of MooA; however, the protease had no effect on fibrinolysis. Our findings show that MooA is a novel hemostatically active metalloprotease, which may lead to the development of coagulopathies during B. moojeni envenomation. Moreover, the metalloprotease may contribute to the development of new diagnostic tools and pharmacological approaches applied to hemostatic disorders.


Asunto(s)
Coagulación Sanguínea/efectos de los fármacos , Bothrops/metabolismo , Coagulantes/farmacología , Venenos de Crotálidos/enzimología , Factor Xa/metabolismo , Leucocitos/efectos de los fármacos , Metaloendopeptidasas/farmacología , Metaloproteasas/farmacología , Protrombina/metabolismo , Tromboplastina/metabolismo , Adulto , Secuencia de Aminoácidos , Animales , Coagulantes/aislamiento & purificación , Venenos de Crotálidos/aislamiento & purificación , Venenos de Crotálidos/farmacología , Estabilidad de Enzimas , Femenino , Humanos , Concentración de Iones de Hidrógeno , Mediadores de Inflamación/metabolismo , Cinética , Leucocitos/metabolismo , Masculino , Metaloendopeptidasas/aislamiento & purificación , Metaloproteasas/aislamiento & purificación , Persona de Mediana Edad , Temperatura , Adulto Joven
6.
Mol Immunol ; 148: 68-80, 2022 08.
Artículo en Inglés | MEDLINE | ID: mdl-35659727

RESUMEN

The successful establishment of HIV-1 infection is related to inflammasome blocking or inactivation, which can result in the viral evasion of the immune responses and formation of reservoirs in several tissues. In this sense, we aimed to evaluate the viral and cellular mechanisms activated during HIV-1 infection in human primary macrophages that allow an effective viral replication in these cells. We found that resting HIV-1-infected macrophages, but not those activated in classical or alternative patterns, released IL-1ß and other pro-inflammatory cytokines, and showed increased CXCL10 expression, without changes in the NLRP3, AIM2 or RIG-I inflammasome pathways. Also, similar levels of Casp-1, phosphorylated NF-κB (p65) and NLRP3 proteins were found in uninfected and HIV-1-infected macrophages. Likewise, no alterations were detected in ASC specks released in the culture supernatant after HIV-1 infection, suggesting that macrophages remain viable after infection. Using in silico prediction studies, we found that the HIV-1 proteins Gag and Vpr interact with several host proteins. Comparable levels of trans-LTB4 were found in the supernatants of uninfected and HIV-1-infected macrophages, whereas ROS production was impaired in infected cells, which was not reversed after the PMA stimulus. Immunofluorescence analysis showed structural alterations in the mitochondrial architecture and an increase of BIM in the cytoplasm of infected cells. Our data suggest that HIV-1 proteins Gag and Vpr, through interacting with cellular proteins in the early steps of infection, preclude the inflammasome activation and the development of effective immune responses, thus allowing the establishment of the infection.


Asunto(s)
Infecciones por VIH , VIH-1 , Infecciones por VIH/metabolismo , Humanos , Inflamasomas , Interleucina-1beta/metabolismo , Macrófagos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Infección Persistente
7.
Sci Adv ; 8(37): eabo5400, 2022 09 16.
Artículo en Inglés | MEDLINE | ID: mdl-36103544

RESUMEN

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induces mild or asymptomatic COVID-19 in most cases, but some patients develop an excessive inflammatory process that can be fatal. As the NLRP3 inflammasome and additional inflammasomes are implicated in disease aggravation, drug repositioning to target inflammasomes emerges as a strategy to treat COVID-19. Here, we performed a high-throughput screening using a 2560 small-molecule compound library and identified FDA-approved drugs that function as pan-inflammasome inhibitors. Our best hit, niclosamide (NIC), effectively inhibits both inflammasome activation and SARS-CoV-2 replication. Mechanistically, induction of autophagy by NIC partially accounts for inhibition of NLRP3 and AIM2 inflammasomes, but NIC-mediated inhibition of NAIP/NLRC4 inflammasome are autophagy independent. NIC potently inhibited inflammasome activation in human monocytes infected in vitro, in PBMCs from patients with COVID-19, and in vivo in a mouse model of SARS-CoV-2 infection. This study provides relevant information regarding the immunomodulatory functions of this promising drug for COVID-19 treatment.


Asunto(s)
Tratamiento Farmacológico de COVID-19 , Inflamasomas , Animales , Humanos , Agentes Inmunomoduladores , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR , SARS-CoV-2
8.
Life Sci ; 285: 119988, 2021 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-34592238

RESUMEN

Strategies capable of attenuating TLR4 can attenuate metabolic processes such as inflammation, endoplasmic reticulum (ER) stress, and apoptosis in the body. Physical exercise has been a cornerstone in suppressing inflammation and dysmetabolic outcomes caused by TRL4 activation. Thus, the present study aimed to evaluate the effects of a chronic physical exercise protocol on the TLR4 expression and its repercussion in the inflammation, ER stress, and apoptosis pathways in mice hearts. Echocardiogram, RT-qPCR, immunoblotting, and histological techniques were used to evaluate the left ventricle of wild-type (WT) and Tlr4 knockout (TLR4 KO) mice submitted to a 4-week physical exercise protocol. Moreover, we performed a bioinformatics analysis to expand the relationship of Tlr4 mRNA in the heart with inflammation, ER stress, and apoptosis-related genes of several isogenic strains of BXD mice. The TLR4 KO mice had higher energy expenditure and heart rate in the control state but lower activation of apoptosis and ER stress pathways. The bioinformatics analysis reinforced these data. In the exercised state, the WT mice improved performance and cardiac function. However, these responses were blunted in the KO group. In conclusion, TLR4 has an essential role in the inhibition of apoptosis and ER stress pathways, as well as in the training-induced beneficial adaptations.


Asunto(s)
Apoptosis/genética , Estrés del Retículo Endoplásmico/genética , Metabolismo Energético/genética , Ventrículos Cardíacos , Condicionamiento Físico Animal , Receptor Toll-Like 4/genética , Función Ventricular , Animales , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Ecocardiografía , Eliminación de Gen , Glucógeno/metabolismo , Frecuencia Cardíaca , Inflamación/genética , Inflamación/patología , Ratones , Ratones Noqueados , ARN Mensajero/metabolismo
9.
Viruses ; 13(12)2021 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-34960790

RESUMEN

Uncontrolled inflammatory responses play a critical role in coronavirus disease (COVID-19). In this context, because the triggering-receptor expressed on myeloid cells-1 (TREM-1) is considered an intrinsic amplifier of inflammatory signals, this study investigated the role of soluble TREM-1 (sTREM-1) as a biomarker of the severity and mortality of COVID-19. Based on their clinical scores, we enrolled COVID-19 positive patients (n = 237) classified into mild, moderate, severe, and critical groups. Clinical data and patient characteristics were obtained from medical records, and their plasma inflammatory mediator profiles were evaluated with immunoassays. Plasma levels of sTREM-1 were significantly higher among patients with severe disease compared to all other groups. Additionally, levels of sTREM-1 showed a significant positive correlation with other inflammatory parameters, such as IL-6, IL-10, IL-8, and neutrophil counts, and a significant negative correlation was observed with lymphocyte counts. Most interestingly, sTREM-1 was found to be a strong predictive biomarker of the severity of COVID-19 and was related to the worst outcome and death. Systemic levels of sTREM-1 were significantly correlated with the expression of matrix metalloproteinases (MMP)-8, which can release TREM-1 from the surface of peripheral blood cells. Our findings indicated that quantification of sTREM-1 could be used as a predictive tool for disease outcome, thus improving the timing of clinical and pharmacological interventions in patients with COVID-19.


Asunto(s)
Biomarcadores/sangre , COVID-19/diagnóstico , COVID-19/mortalidad , Leucocitos/metabolismo , Metaloproteinasa 8 de la Matriz/metabolismo , Índice de Severidad de la Enfermedad , Receptor Activador Expresado en Células Mieloides 1/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Brasil , Femenino , Humanos , Inflamación , Interleucina-10/sangre , Interleucina-6/sangre , Interleucina-8/sangre , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Neutrófilos/metabolismo , Estudios Prospectivos , SARS-CoV-2 , Receptor Activador Expresado en Células Mieloides 1/metabolismo , Adulto Joven
10.
J Exp Med ; 218(3)2021 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-33231615

RESUMEN

Severe cases of COVID-19 are characterized by a strong inflammatory process that may ultimately lead to organ failure and patient death. The NLRP3 inflammasome is a molecular platform that promotes inflammation via cleavage and activation of key inflammatory molecules including active caspase-1 (Casp1p20), IL-1ß, and IL-18. Although participation of the inflammasome in COVID-19 has been highly speculated, the inflammasome activation and participation in the outcome of the disease are unknown. Here we demonstrate that the NLRP3 inflammasome is activated in response to SARS-CoV-2 infection and is active in COVID-19 patients. Studying moderate and severe COVID-19 patients, we found active NLRP3 inflammasome in PBMCs and tissues of postmortem patients upon autopsy. Inflammasome-derived products such as Casp1p20 and IL-18 in the sera correlated with the markers of COVID-19 severity, including IL-6 and LDH. Moreover, higher levels of IL-18 and Casp1p20 are associated with disease severity and poor clinical outcome. Our results suggest that inflammasomes participate in the pathophysiology of the disease, indicating that these platforms might be a marker of disease severity and a potential therapeutic target for COVID-19.


Asunto(s)
COVID-19/patología , COVID-19/virología , Inflamasomas/metabolismo , SARS-CoV-2/fisiología , Índice de Severidad de la Enfermedad , Apoptosis , Comorbilidad , Citocinas/biosíntesis , Humanos , Pulmón/patología , Monocitos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Cambios Post Mortem , Resultado del Tratamiento
11.
J Immunol ; 181(12): 8544-51, 2008 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-19050273

RESUMEN

Leukotrienes (LTs) are potent lipid mediators involved in the control of host defense. LTB(4) induces leukocyte accumulation, enhances phagocytosis and bacterial clearance, and increases NO synthesis. LTB(4) is also important in early effector T cell recruitment that is mediated by LTB(4) receptor 1, the high-affinity receptor for LTB(4). The aims of this study were to evaluate whether LTs are involved in the secondary immune response to vaccination in a murine model of Histoplasma capsulatum infection. Our results demonstrate that protection of wild-type mice immunized with cell-free Ags from H. capsulatum against histoplasmosis was associated with increased LTB(4) and IFN-gamma production as well as recruitment of memory T cells into the lungs. In contrast, cell-free Ag-immunized mice lacking 5-lipoxygenase(-/-), a critical enzyme involved in LT synthesis, displayed a marked decrease on recruitment of memory T cells to the lungs associated with increased synthesis of TGF-beta as well as IL-10. Strikingly, these effects were associated with increased mortality to 5-lipoxygenase(-/-)-infected mice. These data establish an important immunomodulatory role of LTs, in both the primary and secondary immune responses to histoplasmosis.


Asunto(s)
Adyuvantes Inmunológicos/fisiología , Histoplasmosis/inmunología , Inmunización Secundaria , Memoria Inmunológica , Leucotrieno B4/fisiología , Subgrupos de Linfocitos T/inmunología , Adyuvantes Inmunológicos/antagonistas & inhibidores , Adyuvantes Inmunológicos/biosíntesis , Animales , Antígenos Fúngicos/administración & dosificación , Antígenos Fúngicos/inmunología , Movimiento Celular/inmunología , Citocinas/biosíntesis , Vacunas Fúngicas/administración & dosificación , Vacunas Fúngicas/inmunología , Histoplasma/inmunología , Histoplasmosis/microbiología , Histoplasmosis/prevención & control , Humanos , Inmunidad Innata , Leucotrieno B4/antagonistas & inhibidores , Leucotrieno B4/biosíntesis , Enfermedades Pulmonares Fúngicas/inmunología , Enfermedades Pulmonares Fúngicas/patología , Enfermedades Pulmonares Fúngicas/prevención & control , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/microbiología
12.
Am J Respir Crit Care Med ; 180(12): 1227-38, 2009 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-19797157

RESUMEN

RATIONALE: Accumulating evidence supports the hypothesis that the continuous host response to a persistent challenge can polarize the cytokine environment toward a Th2 cytokine phenotype, but the mechanisms responsible for this skewing are not clear. OBJECTIVES: We investigated the role of Toll-like receptor 9 (TLR9) in a Th2-driven pulmonary granulomatous response initiated via the embolization of Schistosoma mansoni eggs to the lungs of mice. METHODS: Mice were intravenously injected with S. mansoni eggs. Histological and flow cytometric analysis, cytokine measurement, adoptive transfer of bone marrow (BM)-derived dendritic cells (DCs), and in vitro T-cell treatments with antigen-presenting cells were examined. MEASUREMENTS AND MAIN RESULTS: In comparison to wild-type mice, TLR9(-/-) mice showed increased pulmonary granuloma size, augmented collagen deposition, increased Th2 cytokine phenotype, and impaired accumulation of DCs. BM-derived DCs, but not macrophages, recovered from animals with developed Th2-type lung granulomas promoted the production of type 2 cytokines from CD4(+) T cells. BM-derived DCs from TLR9(-/-) mice induced impaired Th1 cytokine and enhanced Th2 cytokine production by T cells, compared with DCs from WT mice. Macrophages from TLR9(-/-) mice expressed a significantly higher alternatively activated (M2) phenotype characterized by increased "found in inflammatory zone-1" (FIZZ1) and arginase-1 expression. The adoptive transfer of BM-derived DCs from syngeneic WT mice into TLR9(-/-) mice restored the granuloma phenotype seen in WT mice. CONCLUSIONS: These studies suggest that TLR9 plays an important mechanistic role in the maintenance of the pulmonary granulomatous response.


Asunto(s)
Granuloma del Sistema Respiratorio/inmunología , Inflamación/inmunología , Receptor Toll-Like 9/inmunología , Animales , Enfermedad Crónica , Células Dendríticas/inmunología , Modelos Animales de Enfermedad , Citometría de Flujo , Granuloma del Sistema Respiratorio/complicaciones , Inflamación/complicaciones , Pulmón , Masculino , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Schistosoma mansoni/inmunología
13.
Tuberculosis (Edinb) ; 124: 101980, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-32801053

RESUMEN

Tuberculosis (TB) is the leading cause of mortality among infectious diseases worldwide. The study of molecular targets for therapy and diagnosis suggested that Notch signaling is an important pathway for the maintenance of the immune response during Mycobacterium tuberculosis (Mtb) infection. We evaluated the participation of the Notch pathway in the modulation of immune response during Mtb infection, and observed that patients with active TB had increased DLL4 expression in intermediate and non-classic monocytes. Further, patients with moderate and advanced lung injury have higher Notch1 expression in CD4+ T cells when compared to patients with a minimal lung injury. When we considered the severity of disease in active TB patients, the expression of the DLL4 in intermediate monocytes and the expression of Notch1 in CD4+ T cells are positively correlated with the degree of lung injury. In vitro, PBMCs treated with the Notch pharmacological inhibitor reduced the production of IL-17A and IL-2, whereas anti-hDLL4 treatment promoted a significant increase in TNF-α and phagocytosis. We suggest that Notch1 and DLL4 are associated with immune response activation in human tuberculosis, and can be a novel target to be exploited in the future in the searching of biomarkers.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Linfocitos T CD4-Positivos/metabolismo , Proteínas de Unión al Calcio/metabolismo , Pulmón/metabolismo , Mycobacterium tuberculosis/inmunología , Receptor Notch1/metabolismo , Tuberculosis Pulmonar/metabolismo , Adulto , Biomarcadores/metabolismo , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/microbiología , Estudios de Casos y Controles , Células Cultivadas , Citocinas/metabolismo , Progresión de la Enfermedad , Femenino , Interacciones Huésped-Patógeno , Humanos , Pulmón/inmunología , Pulmón/microbiología , Masculino , Persona de Mediana Edad , Fagocitosis , Índice de Severidad de la Enfermedad , Transducción de Señal , Tuberculosis Pulmonar/inmunología , Tuberculosis Pulmonar/microbiología , Adulto Joven
14.
Life Sci ; 240: 117107, 2020 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-31785241

RESUMEN

BACKGROUND: Toll-like receptor 4 (Tlr4) is recognized due to its role in the immune response. Also, this protein can participate in the signaling pathway of events triggered by physical exercise such as apoptosis, inflammation, and endoplasmic reticulum (ER) stress. The main objective of this study was to evaluate the role of Tlr4 in the markers of these events in the myocardium of mice submitted to acute physical exercise (APE) protocols at different intensities. METHODS: Echocardiogram, RT-qPCR, and immunoblotting technique were used to evaluate the left ventricle of wild-type (WT) and Tlr4 knockout (Tlr4 KO) submitted to APE protocols at 45, 60, and 75% of their maximal velocity. Also, we performed the bioinformatics analysis to establish the connection of heart mRNA levels of Tlr4 with heart genes of inflammation and ER stress of several isogenic strains of BXD mice. RESULTS: Under basal conditions, the Tlr4 deletion diminished the performance, and expression of inflammation and ER stress genes in the left ventricle, but increased the serum levels of CK, Il-17, and Tnf-alpha. Under the same exercise conditions, the Tlr4 deletion reduced the glycemia, serum levels of CK, Il-17, and Tnf-alpha, as well as genes and/or proteins related to apoptosis, inflammation and ER stress in the left ventricle, but increased the levels of CK-mb and LDH, as well as other genes related to apoptosis, inflammation, and ER stress in the left ventricle. CONCLUSION: Altogether, the current findings highlighted the effects of different acute exercise intensities were attenuated in the heart of Tlr4 KO mice.


Asunto(s)
Apoptosis/fisiología , Estrés del Retículo Endoplásmico/fisiología , Corazón/fisiología , Inflamación , Esfuerzo Físico/fisiología , Receptor Toll-Like 4/fisiología , Animales , Apoptosis/genética , Biología Computacional , Creatina Quinasa/sangre , Ecocardiografía , Estrés del Retículo Endoplásmico/genética , Corazón/diagnóstico por imagen , Interleucina-17/sangre , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Esfuerzo Físico/genética , Transducción de Señal/fisiología , Receptor Toll-Like 4/genética , Factor de Necrosis Tumoral alfa/sangre
16.
Toxicol In Vitro ; 61: 104586, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31271808

RESUMEN

The aim of this study was to evaluate the immunomodulatory effects of two toxins from Bothrops snake venoms (the P-I metalloprotease Batroxase and the thrombin-like serine protease Moojase) on human peripheral blood mononuclear cells (PBMC), also investigating changes in the expression of genes related to epigenetic alterations and their immunotherapeutic potential. After 24 h of PBMC stimulation, Batroxase (2 µg/mL) and Moojase (4 µg/mL) increased some cytokine levels (including IL-6 and IL-10), but did not promote cell death processes (apoptosis/necrosis) or alterations in the global DNA methylation levels. Gene expression experiments (RT-qPCR) showed that most of the genes with altered transcript levels encode enzymes that act on histones, such as acetyltransferases (HAT1), deacetylases (HDACs), methyltransferases (DOT1L) or demethylases (KDM5B), indicating that these toxins may alter gene regulation through epigenetic changes mainly related to histones and to methyl-CpG binding proteins (MECP2). Subsequently, the immunotherapeutic potential of these toxins was evaluated using in vitro cytotoxicity assays with NK cells and K562 leukemic cells. Both toxins were able to potentiate the NK cell cytotoxic effects against K562 tumor cells, and the effect of Batroxase was dependent on the concomitant stimulus with IL-2, whereas Moojase increased the NK cytotoxicity independently of IL-2. Thus, Batroxase and Moojase presented interesting immunomodulatory effects that could be explored for the development of new strategies in anticancer immunotherapies.


Asunto(s)
Venenos de Crotálidos/toxicidad , Factores Inmunológicos/toxicidad , Leucocitos Mononucleares/efectos de los fármacos , Metaloproteasas/toxicidad , Proteínas de Reptiles/toxicidad , Adulto , Animales , Bothrops , Supervivencia Celular , Citocinas/metabolismo , Epigénesis Genética/efectos de los fármacos , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Células K562 , Células Asesinas Naturales , Leucocitos Mononucleares/metabolismo , Masculino , Adulto Joven
17.
J Endocrinol ; 240(2): 181-193, 2019 02 01.
Artículo en Inglés | MEDLINE | ID: mdl-30400033

RESUMEN

The endoplasmic reticulum (ER) stress and inflammation relationship occurs at different levels and is essential for the adequate homeostatic function of cellular systems, becoming harmful when chronically engaged. Intense physical exercise enhances serum levels of interleukin 6 (IL-6). In response to a chronic exhaustive physical exercise protocol, our research group verified an increase of the IL-6 concentration and ER stress proteins in extensor digitorium longus (EDL) and soleus. Based on these results, we hypothesized that IL-6-knockout mice would demonstrate a lower modulation in the ER stress proteins compared to the wild-type mice. To clarify the relationship between exercise-induced IL-6 increased and ER stress, we studied the effects of an acute exhaustive physical exercise protocol on the levels of ER stress proteins in the skeletal muscles of IL-6-knockout (KO) mice. The WT group displayed a higher exhaustion time compared to the IL-6 KO group. After 1 h of the acute exercise protocol, the serum levels of IL-6 and IL-10 were enhanced in the WT group. Independent of the experimental group, the CHOP and cleaved caspase 12/total caspase 12 ratio in EDL as well as ATF6 and CHOP in soleus were sensitive to the acute exercise protocol. Compared to the WT group, the oscillation patterns over time of BiP in EDL and soleus as well as of peIF2-alpha/eIF2-alpha ratio in soleus were attenuated for the IL-6 KO group. In conclusion, IL-6 seems to be related with the ER stress homeostasis, once knockout mice presented attenuation of BiP in EDL and soleus as well as of pEiF2-alpha/EiF2-alpha ratio in soleus after the acute exhaustive physical exercise protocol.


Asunto(s)
Estrés del Retículo Endoplásmico/fisiología , Interleucina-6/metabolismo , Músculo Esquelético/metabolismo , Condicionamiento Físico Animal/fisiología , Factor de Transcripción Activador 6/metabolismo , Animales , Western Blotting , Caspasas/metabolismo , Estrés del Retículo Endoplásmico/genética , Interleucina-10/sangre , Interleucina-10/metabolismo , Interleucina-6/sangre , Interleucina-6/genética , Ratones Endogámicos C57BL , Ratones Noqueados , Factor de Transcripción CHOP/metabolismo
18.
Nat Commun ; 10(1): 5273, 2019 11 21.
Artículo en Inglés | MEDLINE | ID: mdl-31754185

RESUMEN

Leishmania RNA virus (LRV) is an important virulence factor associated with the development of mucocutaneous Leishmaniasis, a severe form of the disease. LRV-mediated disease exacerbation relies on TLR3 activation, but downstream mechanisms remain largely unexplored. Here, we combine human and mouse data to demonstrate that LRV triggers TLR3 and TRIF to induce type I IFN production, which induces autophagy. This process results in ATG5-mediated degradation of NLRP3 and ASC, thereby limiting NLRP3 inflammasome activation in macrophages. Consistent with the known restricting role of NLRP3 for Leishmania replication, the signaling pathway triggered by LRV results in increased parasite survival and disease progression. In support of this data, we find that lesions in patients infected with LRV+ Leishmania are associated with reduced inflammasome activation and the development of mucocutaneous disease. Our findings reveal the mechanisms triggered by LRV that contribute to the development of the debilitating mucocutaneous form of Leishmaniasis.


Asunto(s)
Inmunidad Innata/inmunología , Inflamasomas/inmunología , Leishmania/inmunología , Leishmaniasis Mucocutánea/inmunología , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Virus ARN/inmunología , Receptor Toll-Like 3/inmunología , Animales , Autofagia/inmunología , Humanos , Interleucina-1beta/inmunología , Interleucina-1beta/metabolismo , Leishmania/fisiología , Leishmania/virología , Leishmaniasis Mucocutánea/parasitología , Leishmaniasis Mucocutánea/virología , Macrófagos/inmunología , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Virus ARN/fisiología , Transducción de Señal/inmunología , Receptor Toll-Like 3/metabolismo
19.
BMC Immunol ; 9: 38, 2008 Jul 22.
Artículo en Inglés | MEDLINE | ID: mdl-18647414

RESUMEN

BACKGROUND: The greatest challenges in vaccine development include optimization of DNA vaccines for use in humans, creation of effective single-dose vaccines, development of delivery systems that do not involve live viruses, and the identification of effective new adjuvants. Herein, we describe a novel, simple technique for efficiently vaccinating mice against tuberculosis (TB). Our technique consists of a single-dose, genetic vaccine formulation of DNA-hsp65 complexed with cationic liposomes and administered intranasally. RESULTS: We developed a novel and non-toxic formulation of cationic liposomes, in which the DNA-hsp65 vaccine was entrapped (ENTR-hsp65) or complexed (COMP-hsp65), and used to immunize mice by intramuscular or intranasal routes. Although both liposome formulations induced a typical Th1 pattern of immune response, the intramuscular route of delivery did not reduce the number of bacilli. However, a single intranasal immunization with COMP-hsp65, carrying as few as 25 microg of plasmid DNA, leads to a remarkable reduction of the amount of bacilli in lungs. These effects were accompanied by increasing levels of IFN-gamma and lung parenchyma preservation, results similar to those found in mice vaccinated intramuscularly four times with naked DNA-hsp65 (total of 400 microg). CONCLUSION: Our objective was to overcome the significant obstacles currently facing DNA vaccine development. Our results in the mouse TB model showed that a single intranasal dose of COMP-hsp65 elicited a cellular immune response that was as strong as that induced by four intramuscular doses of naked-DNA. This formulation allowed a 16-fold reduction in the amount of DNA administered. Moreover, we demonstrated that this vaccine is safe, biocompatible, stable, and easily manufactured at a low cost. We believe that this strategy can be applied to human vaccines to TB in a single dose or in prime-boost protocols, leading to a tremendous impact on the control of this infectious disease.


Asunto(s)
Proteínas Bacterianas/administración & dosificación , Chaperoninas/administración & dosificación , Mycobacterium tuberculosis , Vacunas contra la Tuberculosis/administración & dosificación , Tuberculosis Pulmonar/inmunología , Vacunas de ADN/administración & dosificación , Administración Intranasal , Animales , Proteínas Bacterianas/inmunología , Chaperonina 60 , Chaperoninas/inmunología , Femenino , Inmunidad Activa/efectos de los fármacos , Inmunización Secundaria , Liposomas , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Células TH1/efectos de los fármacos , Tuberculosis Pulmonar/microbiología , Tuberculosis Pulmonar/prevención & control
20.
Int J Biol Macromol ; 109: 664-671, 2018 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-29274419

RESUMEN

Snake venoms are complex mixtures of organic and inorganic compounds, including proteins belonging to the protease (serine and metalloproteinases), oxidase (L-amino acid oxidases), and phospholipase (especially phospholipases A2) enzyme classes. These toxins account for the serious deleterious effects of snake envenomations, such as tissue necrosis, neurotoxicity, and hemorrhage. In addition to their toxic effects, snake venom toxins have served as important tools for investigating the mechanisms underlying envenomation and discovering new pharmacologically active compounds with immunotherapeutic potential. In this sense, the present review discusses the new findings and therapeutic perspectives in the immune modulating potential of enzymatic toxins from snake venoms belonging to the classes metalloproteinase, serine protease, L-amino acid oxidase, and phospholipase A2.


Asunto(s)
Enzimas/química , Enzimas/metabolismo , Venenos de Serpiente/química , Venenos de Serpiente/enzimología , Toxinas Biológicas/química , Toxinas Biológicas/metabolismo , Animales , Enzimas/inmunología , Humanos , Inmunomodulación , Mordeduras de Serpientes/inmunología , Mordeduras de Serpientes/metabolismo , Mordeduras de Serpientes/patología , Mordeduras de Serpientes/terapia , Venenos de Serpiente/inmunología , Venenos de Serpiente/uso terapéutico , Toxinas Biológicas/inmunología
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