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1.
Cytometry B Clin Cytom ; 100(1): 33-41, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33394568

RESUMEN

Over a remarkably short period of time, a great deal of knowledge about severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection has been acquired, through the focused and cooperative effort of the international scientific community. Much has become known about how the immune response is coordinated to fight infection, and how it becomes dysregulated in severe disease. In this review, we take an in-depth look at the many immune features associated with the host response to SARS-CoV2, as well as those that appear to mark severe disease.


Asunto(s)
/diagnóstico por imagen , Citometría de Flujo/métodos , Técnica del Anticuerpo Fluorescente/métodos , /inmunología , Biomarcadores/análisis , /terapia , Quimiocinas/análisis , Quimiocinas/metabolismo , Citocinas/análisis , Citocinas/metabolismo , Técnica del Anticuerpo Fluorescente/tendencias , Interacciones Huésped-Patógeno/inmunología , Humanos , Inmunidad/fisiología , Metabolómica/métodos , Metabolómica/tendencias , Medición de Riesgo , Índice de Severidad de la Enfermedad
2.
BMC Infect Dis ; 20(1): 505, 2020 Jul 13.
Artículo en Inglés | MEDLINE | ID: mdl-32660552

RESUMEN

BACKGROUND: Meningococcal meningitis (MM) is a life-threatening disease associated with approximately 10% case fatality rates and neurological sequelae in 10-20% of the cases. Recently, we have shown that the matrix metalloproteinase (MMP) inhibitor BB-94 reduced brain injury in a mouse model of MM. The present study aimed to assess whether doxycycline (DOX), a tetracycline that showed a neuroprotective effect as adjuvant therapy in experimental pneumococcal meningitis (PM), would also exert a beneficial effect when given as adjunctive therapy to ceftriaxone (CRO) in experimental MM. METHODS: BALB/c mice were infected by the intracisternal route with a group C Neisseria meningitidis strain. Eighteen h post infection (hpi), animals were randomised for treatment with CRO [100 mg/kg subcutaneously (s.c.)], CRO plus DOX (30 mg/kg s.c.) or saline (control s.c.). Antibiotic treatment was repeated 24 and 40 hpi. Mouse survival and clinical signs, bacterial counts in cerebella, brain damage, MMP-9 and cyto/chemokine levels were assessed 48 hpi. RESULTS: Analysis of bacterial load in cerebella indicated that CRO and CRO + DOX were equally effective at controlling meningococcal replication. No differences in survival were observed between mice treated with CRO (94.4%) or CRO + DOX (95.5%), (p > 0.05). Treatment with CRO + DOX significantly diminished both the number of cerebral hemorrhages (p = 0.029) and the amount of MMP-9 in the brain (p = 0.046) compared to untreated controls, but not to CRO-treated animals (p > 0.05). Levels of inflammatory markers in the brain of mice that received CRO or CRO + DOX were not significantly different (p > 0.05). Overall, there were no significant differences in the parameters assessed between the groups treated with CRO alone or CRO + DOX. CONCLUSIONS: Treatment with CRO + DOX showed similar bactericidal activity to CRO in vivo, suggesting no antagonist effect of DOX on CRO. Combined therapy significantly improved mouse survival and disease severity compared to untreated animals, but addition of DOX to CRO did not offer significant benefits over CRO monotherapy. In contrast to experimental PM, DOX has no adjunctive activity in experimental MM.


Asunto(s)
Antibacterianos/uso terapéutico , Ceftriaxona/uso terapéutico , Doxiciclina/uso terapéutico , Meningitis Meningocócica/tratamiento farmacológico , Neisseria meningitidis Serogrupo C , Animales , Antibacterianos/administración & dosificación , Carga Bacteriana/efectos de los fármacos , Ceftriaxona/administración & dosificación , Hemorragia Cerebral/tratamiento farmacológico , Quimiocinas/análisis , Quimiocinas/metabolismo , Modelos Animales de Enfermedad , Doxiciclina/administración & dosificación , Quimioterapia Combinada , Femenino , Humanos , Estimación de Kaplan-Meier , Metaloproteinasa 9 de la Matriz/análisis , Metaloproteinasa 9 de la Matriz/metabolismo , Meningitis Meningocócica/mortalidad , Ratones , Ratones Endogámicos BALB C , Distribución Aleatoria , Resultado del Tratamiento
3.
J Surg Res ; 254: 75-82, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32417499

RESUMEN

BACKGROUND: The use of mesenchymal stem cells (MSCs) for treatment during ischemia is novel. Hydrogen sulfide (H2S) is an important paracrine mediator that is released from MSCs to facilitate angiogenesis and vasodilation. Three enzymes, cystathionine-beta-synthase (CBS), cystathionine-gamma-lyase (CSE), and 3-mercaptopyruvate-sulfurtransferase (MPST), are mainly responsible for H2S production. However, it is unclear how these enzymes impact the production of other critical growth factors and chemokines. We hypothesized that the enzymes responsible for H2S production in human MSCs would also critically regulate other growth factors and chemokines. MATERIALS AND METHODS: Human MSCs were transfected with CBS, MPST, CSE, or negative control small interfering RNA. Knockdown of enzymes was confirmed by polymerase chain reaction. Cells were plated in 12-well plates at 100,000 cells per well and stimulated with tumor necrosis factor-α (TNF-α; 50 ng/mL), lipopolysaccharide (LPS; 200 ng/mL), or 5% hypoxia for 24 h. Supernatants were collected, and cytokines measured by multiplex beaded assay. Data were compared with the Mann-Whitney U-test, and P < 0.05 was significant. RESULTS: TNF-α, LPS, and hypoxia effectively stimulated MSCs. Granulocyte colony-stimulating factor (GCSF), epidermal growth factor, fibroblast growth factor, granulocyte/monocyte colony-stimulating factor (GMCSF), vascular endothelial growth factor, and interferon gamma-inducible protein 10 were all significantly elevated when CSE was knocked down during TNF-α stimulation (P < 0.05). Knockdown of MPST during LPS stimulation more readily increased GCSF and epidermal growth factor but decreased GMCSF (P < 0.05). CBS knockdown decreased production of GCSF, fibroblast growth factor, GMCSF, and vascular endothelial growth factor (P < 0.05) after hypoxia. CONCLUSIONS: The enzymes that produce H2S in MSCs are also responsible for the production of other stem cell paracrine mediators under stressful stimuli. Therefore, reprogramming MSCs to endogenously produce more H2S as a therapeutic intervention could also critically impact other paracrine mediators, which may alter the desired beneficial effects.


Asunto(s)
Sulfuro de Hidrógeno/metabolismo , Células Madre Mesenquimatosas/metabolismo , Comunicación Paracrina/fisiología , Hipoxia de la Célula , Células Cultivadas , Quimiocinas/análisis , Quimiocinas/metabolismo , Cistationina betasintasa/genética , Cistationina betasintasa/fisiología , Cistationina gamma-Liasa/genética , Cistationina gamma-Liasa/fisiología , Técnicas de Silenciamiento del Gen , Humanos , Sulfuro de Hidrógeno/farmacología , Péptidos y Proteínas de Señalización Intercelular/análisis , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Lipopolisacáridos/farmacología , Comunicación Paracrina/efectos de los fármacos , Sulfurtransferasas/genética , Sulfurtransferasas/fisiología , Transfección , Factor de Necrosis Tumoral alfa/farmacología
4.
Emerg Microbes Infect ; 9(1): 761-770, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32228226

RESUMEN

Circulating in China and 158 other countries and areas, the ongoing COVID-19 outbreak has caused devastating mortality and posed a great threat to public health. However, efforts to identify effectively supportive therapeutic drugs and treatments has been hampered by our limited understanding of host immune response for this fatal disease. To characterize the transcriptional signatures of host inflammatory response to SARS-CoV-2 (HCoV-19) infection, we carried out transcriptome sequencing of the RNAs isolated from the bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells (PBMC) specimens of COVID-19 patients. Our results reveal distinct host inflammatory cytokine profiles to SARS-CoV-2 infection in patients, and highlight the association between COVID-19 pathogenesis and excessive cytokine release such as CCL2/MCP-1, CXCL10/IP-10, CCL3/MIP-1A, and CCL4/MIP1B. Furthermore, SARS-CoV-2 induced activation of apoptosis and P53 signalling pathway in lymphocytes may be the cause of patients' lymphopenia. The transcriptome dataset of COVID-19 patients would be a valuable resource for clinical guidance on anti-inflammatory medication and understanding the molecular mechansims of host response.


Asunto(s)
Líquido del Lavado Bronquioalveolar , Quimiocinas/análisis , Infecciones por Coronavirus/genética , Citocinas/análisis , Leucocitos Mononucleares , Neumonía Viral/genética , Transcriptoma , Apoptosis , Betacoronavirus , Infecciones por Coronavirus/sangre , Infecciones por Coronavirus/inmunología , Humanos , Linfopenia , Pandemias , Neumonía Viral/sangre , Neumonía Viral/inmunología , RNA-Seq , Transducción de Señal , Proteína p53 Supresora de Tumor
5.
PLoS One ; 15(2): e0229251, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32092101

RESUMEN

Since chemerin's identification as an adipokine, it has been associated with a number of human diseases including diabetes and obesity. However, the basic scientific foundation for these clinical determinations is still lacking. Fibroblastic mouse 3T3 cells are unable to develop lipid droplets if chemerin is not present. Thus, we hypothesized that an in vivo rat model chemerin knockout (KO; an advancement from the previously mentioned in vitro cultures) would have limited accumulation of lipid in adipocytes compared to their wild-type (WT) counterparts. Female WT/KO rats (Sprague Dawley background) were fed a low-fat diet starting at 8 weeks of age with weekly body weight and food consumption monitoring. At 25 weeks of age, adipose tissue depots were dissected and flash frozen for PCR analysis or fixed with paraformaldehyde for histology. Over the 17 weeks of experimentation, WT and KO animals did not have differences in total body weight or food consumption but KO animals had a significantly reduced amount of visceral fat compared to WT animals (via microCT at 8 and 25 weeks). Histology of retroperitoneal and mesenteric depots demonstrated a significant leftward shift in adipocyte size in the mesenteric but not the retroperitoneal depot of the KO compared to WT animals. Similarly, in the mesenteric fat of the KO rat, gene expression of adiponectin, fatty acid synthase, perilipin, and leptin were significantly reduced compared to mesenteric fat of WT animals and retroperitoneal fat of both WT and KO animals. Adiponectin was highlighted by a protein-protein interaction network as being important for the physiological effects of chemerin removal. These data are the first, to our knowledge, to demonstrate chemerin's adipokine potential in vivo and identify it as fat depot location-specific.


Asunto(s)
Adipogénesis/genética , Quimiocinas/análisis , Adipocitos/fisiología , Adipoquinas/fisiología , Tejido Adiposo/citología , Animales , Peso Corporal , Quimiocinas/genética , Quimiocinas/fisiología , Dieta con Restricción de Grasas , Técnicas de Inactivación de Genes , Grasa Intraabdominal , Gotas Lipídicas , Mesenterio/citología , Ratas , Ratas Sprague-Dawley
6.
Chem Biol Interact ; 322: 108968, 2020 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-32004530

RESUMEN

Osteoarthritis (OA) is one of the most prevalent degenerative joint diseases, and the risk of developing OA significantly increases with age as well as with concomitant diseases, such as diabetes. Advanced glycation end products (AGEs) accumulate in the body over time and are associated with increased expression of various molecules involved in the pathophysiology of OA. Prostaglandin E2 (PGE2), along with its precursor cyclooxygenase (COX)-2, plays an integral role in the pathogenesis of OA and is highly upregulated in response to AGEs. The most significant event in OA is excessive degradation of the cartilage extracellular matrix, which is composed primarily of type II collagen and aggrecan. In the present study, we investigated the involvement of the receptor for glucagon-like peptide (GLP)-1 in the response of chondrocytes to insult from AGEs using the selective GLP-1 agonist dulaglutide. Firstly, our results indicate that AGEs reduced the expression of the receptor for GLP-1 (GLP-1R) in human SW1353 chondrocytes. Interestingly, we found that treatment with dulaglutide could ameliorate deterioration of the components of the articular extracellular matrix (ECM), such as type II collagen and aggrecan, induced by AGEs through downregulation of matrix metalloproteinase (MMP)-3 and MMP-13 and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5. We also found that dulaglutide exerted a potent inhibitory effect against the expression of several proinflammatory cytokines and chemokines closely associated with OA, as well as the production of reactive oxygen species (ROS). Finally, we showed that the effects of dulaglutide were mediated through the nuclear factor kappa-B (NF-κB) pathway. Our findings indicate that dulaglutide displayed a robust protective effect against AGEs-induced damage in chondrocytes, suggesting that it might be a possible therapeutic agent for the treatment of OA.


Asunto(s)
Agrecanos/metabolismo , Colágeno Tipo II/metabolismo , Péptidos Similares al Glucagón/análogos & derivados , Productos Finales de Glicación Avanzada/metabolismo , Fragmentos Fc de Inmunoglobulinas/farmacología , Proteolisis/efectos de los fármacos , Proteínas Recombinantes de Fusión/farmacología , Línea Celular , Quimiocinas/análisis , Quimiocinas/metabolismo , Condrocitos/citología , Condrocitos/metabolismo , Citocinas/análisis , Citocinas/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Receptor del Péptido 1 Similar al Glucagón/genética , Receptor del Péptido 1 Similar al Glucagón/metabolismo , Péptidos Similares al Glucagón/farmacología , Humanos , Metaloproteinasa 13 de la Matriz/genética , Metaloproteinasa 13 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/genética , Metaloproteinasa 3 de la Matriz/metabolismo , FN-kappa B/metabolismo , Osteoartritis/metabolismo , Osteoartritis/patología , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo
7.
Blood Adv ; 4(2): 367-379, 2020 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-31985806

RESUMEN

Acute myeloid leukemia (AML) is a genetically heterogeneous disease that is characterized by abnormal clonal proliferation of myeloid progenitor cells found predominantly within the bone marrow (BM) and blood. Recent studies suggest that genetic and phenotypic alterations in the BM microenvironment support leukemogenesis and allow leukemic cells to survive and evade chemotherapy-induced death. However, despite substantial evidence indicating the role of tumor-host interactions in AML pathogenesis, little is known about the complex microenvironment of the BM. To address this, we performed novel proteomic profiling of the noncellular compartment of the BM microenvironment in patients with AML (n = 10) and age- and sex-matched healthy control subjects (n = 10) using an aptamer-based, highly multiplexed, affinity proteomics platform (SOMAscan). We show that proteomic assessment of blood or RNA-sequencing of BM are suboptimal alternate screening strategies to determine the true proteomic composition of the extracellular soluble compartment of AML patient BM. Proteomic analysis revealed that 168 proteins significantly differed in abundance, with 91 upregulated and 77 downregulated in leukemic BM. A highly connected signaling network of cytokines and chemokines, including IL-8, was found to be the most prominent proteomic signature associated with AML in the BM microenvironment. We report the first description of significantly elevated levels of the myelosuppressive chemokine CCL23 (myeloid progenitor inhibitory factor-1) in both AML and myelodysplastic syndrome patients and perform functional experiments supportive of a role in the suppression of normal hematopoiesis. This unique paired RNA-sequencing and proteomics data set provides innovative mechanistic insights into AML and healthy aging and should serve as a useful public resource.


Asunto(s)
Médula Ósea/patología , Leucemia Mieloide Aguda/patología , Proteómica/métodos , Estudios de Casos y Controles , Microambiente Celular , Quimiocinas/análisis , Quimiocinas CC/metabolismo , Citocinas/análisis , Regulación Leucémica de la Expresión Génica , Humanos , Interleucina-8/metabolismo , Proteínas de Neoplasias/análisis
8.
Dis Markers ; 2020: 2696317, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31998415

RESUMEN

Introduction. Exhaled breath condensate (EBC) is a noninvasive method to collect samples from the respiratory tract. Usually, a thermoelectric cooling module is required to collect sufficient EBC volume for analyses. In here, we assessed the feasibility of cytokine and chemokine detection in EBC collected directly from the ventilator circuit without the use of a cooling module: swivel-derived exhaled breath condensate (SEBC). Methods: SEBC was prospectively collected from the swivel adapter and stored at -80°C. The objective of this study was to detect cytokines and chemokines in SEBC with a multiplex immunoassay. Secondary outcomes were to assess the correlation between cytokine and chemokine concentrations in SEBC and mechanical ventilation parameters, systemic inflammation parameters, and hemodynamic parameters. Results: Twenty-nine SEBC samples were obtained from 13 ICU patients. IL-1ß, IL-4, IL-8, and IL-17 were detected in more than 90% of SEBC samples, and significant correlations between multiple cytokines and chemokines were found. Several significant correlations were found between cytokines and chemokines in SEBC and mechanical ventilation parameters and serum lactate concentrations. Conclusion: This pilot study showed that it is feasible to detect cytokines and chemokines in SEBC samples obtained without a cooling module. Despite small sample size, correlations were found between cytokines and chemokines in SEBC and mechanical ventilation parameters, as well as serum lactate concentrations. This simple SEBC collection method provides the opportunity to collect EBC samples in large prospective ICU cohorts.


Asunto(s)
Quimiocinas/análisis , Interleucinas/análisis , /diagnóstico , Adulto , Anciano , Biomarcadores/análisis , Pruebas Respiratorias/instrumentación , Pruebas Respiratorias/métodos , Femenino , Humanos , Inmunoensayo/métodos , Masculino , Persona de Mediana Edad , Ventiladores Mecánicos
9.
Brain Dev ; 42(2): 185-191, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31787380

RESUMEN

BACKGROUND: The pathogenesis of acute encephalopathy (AE) remains unclear, and a biomarker has not been identified. METHODS: Levels of 49 cytokines and chemokines, including osteopontin (OPN), were measured in serum and cerebrospinal fluid (CSF) of children with AE (n = 17) or febrile convulsion (FC; n = 8; control group). The AE group included acute necrotizing encephalopathy (n = 1), acute encephalopathy with biphasic seizures and late reduced diffusion (AESD; n = 3), clinically mild encephalitis/encephalopathy with a reversible splenial lesion (MERS; n = 4), and unclassified acute encephalopathy (UCAE; n = 9) that does not meet the criteria of syndrome classification. Five individuals with AE had neurological sequelae or death (poor prognosis), whereas 12 were alive without neurological sequelae (good prognosis). RESULTS: The CSF:serum ratios of OPN, CC chemokine ligand (CCL)4, and interleukin (IL)-10 were significantly higher in AE than in FC. The CSF levels of macrophage inhibitory factor (MIF) and leukemia inhibitory factor (LIF) were significantly higher in the poor-prognosis group than in the good-prognosis group. The CSF:serum ratios of OPN were significantly higher in AESD and in MERS than in FC. The CSF:serum ratios of MIF and OPN were higher in MERS than in UCAE or FC. CONCLUSION: Our results suggest that microglia-related cytokines and chemokines such as OPN, MIF, and LIF could be novel biomarkers of AE, in addition to the previously reported IL-10 and CCL4, and that MIF and LIF may be markers of poor prognosis.


Asunto(s)
Encefalopatías/inmunología , Encefalopatías/patología , Citocinas/análisis , Biomarcadores/sangre , Biomarcadores/líquido cefalorraquídeo , Quimiocinas/análisis , Quimiocinas/sangre , Quimiocinas/líquido cefalorraquídeo , Preescolar , Citocinas/sangre , Citocinas/líquido cefalorraquídeo , Encefalitis/líquido cefalorraquídeo , Femenino , Humanos , Lactante , Oxidorreductasas Intramoleculares/sangre , Oxidorreductasas Intramoleculares/líquido cefalorraquídeo , Factor Inhibidor de Leucemia/sangre , Factor Inhibidor de Leucemia/líquido cefalorraquídeo , Factores Inhibidores de la Migración de Macrófagos/sangre , Factores Inhibidores de la Migración de Macrófagos/líquido cefalorraquídeo , Masculino , Osteopontina/sangre , Osteopontina/líquido cefalorraquídeo , Convulsiones/etiología , Convulsiones Febriles/complicaciones , Convulsiones Febriles/inmunología , Convulsiones Febriles/patología
10.
J Neurochem ; 152(6): 697-709, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-31782806

RESUMEN

Systemic inflammation can exacerbate symptoms of many neurological diseases. This effect may be facilitated by glial cells of the central nervous system (CNS) that alter their transcriptional responses and up-regulate cytokine and chemokine expression which can, in turn trigger immune surveillance. In this study, we sought to determine the effects of pro-inflammatory cytokine stimulation (TNF, IL-1α, IL-1ß) on astrocyte and microglia chemokine secretion. Primary cultures of astrocytes or microglia were stimulated with the recombinant cytokines and the levels of secreted chemokines were semi-quantitatively determined using a chemokine-specific proteome profiler array and densitometry. Pharmacological inhibitors were used to determine the effects of p38 MAPK, JNK, ERK1/2, NFkB, and transforming growth factor beta-associated kinase 1 (TAK1) in controlling chemokine production. Finally, neutrophil migration assays were performed to demonstrate functionality. Our data show that stimulated astrocytes secrete at least eight chemokines as a response to cytokine stimulation. These include those involved in neutrophil chemo-attraction and proved capable of promoting neutrophil migration in vitro. In contrast, microglia up-regulated few chemokines in response to cytokine stimulation and did not promote neutrophil migration. However, microglia readily secreted chemokines following stimulation with the toll-like receptor agonists. Finally, we show that both the production of chemokines and neutrophil migration resulting from cytokine stimulation of astrocytes was dependent on TAK1 signaling. Collectively, this study adds to the understanding of how astrocytes and microglia respond to stimuli and their role in promoting neutrophil migration to the CNS during inflammatory conditions.


Asunto(s)
Astrocitos/fisiología , Movimiento Celular/fisiología , Quimiocinas/metabolismo , Citocinas/farmacología , Quinasas Quinasa Quinasa PAM/fisiología , Animales , Astrocitos/enzimología , Células Cultivadas , Quimiocinas/análisis , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Femenino , Inflamación/fisiopatología , Lipopolisacáridos/farmacología , Quinasas Quinasa Quinasa PAM/antagonistas & inhibidores , Masculino , Ratones , Ratones Endogámicos C57BL , Microglía/efectos de los fármacos , Microglía/fisiología , Neutrófilos/fisiología , Transducción de Señal/fisiología
11.
Biosci Biotechnol Biochem ; 84(1): 208-215, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31532348

RESUMEN

Dihomo-γ-linolenic acid (DGLA, C20: 3n-6) is known to have an anti-inflammatory activity, but its range of effects was not well studied because of its limited natural sources. We addressed these issues by constructing an yeast Saccharomyces cerevisiae strain having a complete metabolic pathway for DGLA synthesis by introducing two desaturase and one elongase genes to convert endogenous oleic acid to DGLA. Taking advantage of well-known safety of S. cerevisiae, we previously investigated the efficacy of heat-killed whole DGLA-producing yeast cells on irritant contact dermatitis, and showed that oral intake of this yeast significantly suppressed inflammatory reactions, whereas no such suppression was observed by the intake of 25 times the amount of purified DGLA. Since this method is considered to be a simple and efficient way to suppress inflammation, we examined its effectiveness against allergic contact dermatitis (ACD) in this study and showed that this method was also effective against ACD.


Asunto(s)
Ácido 8,11,14-Eicosatrienoico/farmacología , Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Dermatitis Alérgica por Contacto/terapia , Saccharomyces cerevisiae/citología , Saccharomyces cerevisiae/metabolismo , Ácido 8,11,14-Eicosatrienoico/administración & dosificación , Ácido 8,11,14-Eicosatrienoico/metabolismo , Acetona/química , Administración Oral , Animales , Quimiocina CCL2/análisis , Quimiocinas/análisis , Dermatitis Alérgica por Contacto/etiología , Dinitrofluorobenceno/efectos adversos , Dinitrofluorobenceno/inmunología , Oído Externo/patología , Femenino , Inmunización , Inflamación/terapia , Interferón gamma/análisis , Ratones , Ácido Oléico/metabolismo , Aceite de Oliva/química
12.
Sex Transm Infect ; 96(1): 3-9, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31197065

RESUMEN

OBJECTIVES: Recent studies have identified vaginal bacterial taxa associated with increased HIV risk. A possible mechanism to explain these results is that individual taxa differentially promote cervicovaginal inflammation. This study aimed to explore relationships between concentrations of bacteria previously linked to HIV acquisition and vaginal concentrations of proinflammatory cytokines and chemokines. METHODS: In this cross-sectional analysis, concentrations of 17 bacterial taxa and four proinflammatory cytokines (interleukin (IL)-1ß, IL-6, IL-10 and tumour necrosis factor alpha (TNFα)) and two proinflammatory chemokines (IL-8 and interferon gamma-induced protein 10) were measured in vaginal swabs collected from 80 HIV-uninfected women. Cytokine and chemokine concentrations were compared between women with bacterial concentrations above or below the lower limit of detection as determined by quantitative PCR for each taxon. Principal component analysis was used to create a summary score for closely correlated bacteria, and linear regression analysis was used to evaluate associations between this score and increasing concentrations of TNFα and IL-1ß. RESULTS: Detection of Dialister micraerophilus (p=0.01), Eggerthella sp type 1 (p=0.05) or Mycoplasma hominis (p=0.03) was associated with higher TNFα concentrations, and detection of D. micraerophilus (p<0.01), Eggerthella sp type 1 (p=0.04), M. hominis (p=0.02) or Parvimonas sp type 2 (p=0.05) was associated with significantly higher IL-1ß concentrations. Seven bacterial taxa (D. micraerophilus, Eggerthella sp type 1, Gemella asaccharolytica, Sneathia sp, Megasphaera sp, M. hominis and Parvimonas sp type 2) were found to be highly correlated by principal component analysis (eigenvalue 5.24, explaining 74.92% of variability). Linear regression analysis demonstrated associations between this principal component and concentrations of TNFα (ß=0.55, 95% CI 0.01 to 1.08; p=0.048) and IL-1ß (ß=0.96, 95% CI 0.19 to 1.74; p=0.016). CONCLUSIONS: This study provides evidence that several highly correlated vaginal bacterial taxa may influence vaginal cytokine and chemokine concentrations. These results suggest a mechanism where the presence of specific bacterial taxa could influence HIV susceptibility by increasing vaginal inflammation.


Asunto(s)
Bacterias/aislamiento & purificación , Quimiocinas/análisis , Citocinas/análisis , Infecciones por VIH/diagnóstico , Vagina/microbiología , Adolescente , Adulto , Bacterias/clasificación , Bacterias/genética , Quimiocinas/inmunología , Estudios Transversales , Citocinas/inmunología , Susceptibilidad a Enfermedades/diagnóstico , Susceptibilidad a Enfermedades/inmunología , Susceptibilidad a Enfermedades/virología , Femenino , Infecciones por VIH/inmunología , Infecciones por VIH/virología , VIH-1/fisiología , Humanos , Interleucina-1beta/análisis , Interleucina-1beta/inmunología , Microbiota , Persona de Mediana Edad , Factores de Riesgo , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/inmunología , Vagina/química , Vagina/inmunología , Adulto Joven
13.
Sci Rep ; 9(1): 17754, 2019 11 28.
Artículo en Inglés | MEDLINE | ID: mdl-31780824

RESUMEN

Actinic Keratosis (AK), Intraepidermal Carcinoma (IEC), and Squamous Cell Carcinoma (SCC) are generally considered to be advancing stages of the same disease spectrum. However, while AK often regress spontaneously, and IEC often regress in response to immune-activating treatments, SCC typically do not regress. Therefore, it is vital to define whether fundamental immunological changes occur during progression to SCC. Here we show that proinflammatory cytokine expression, chemokine expression, and immune cell infiltration density change during progression to SCC. Our findings suggest a switch from predominantly proinflammatory cytokine production to chemokine production is a key feature of progression from precancer to cancer. Together, these observations propose a model that can underpin current research and open new avenues of exploration into the clinical significance of these profiles with respect to immunotherapeutic or other treatment outcomes.


Asunto(s)
Carcinoma de Células Escamosas/patología , Quimiocinas/análisis , Citocinas/análisis , Neoplasias Cutáneas/patología , Anciano , Anciano de 80 o más Años , Carcinoma in Situ/patología , Progresión de la Enfermedad , Femenino , Humanos , Queratosis Actínica/patología , Masculino , Persona de Mediana Edad , Lesiones Precancerosas/patología , Piel/patología
14.
Methods Enzymol ; 629: 151-176, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31727238

RESUMEN

Over the past two decades there have been tremendous advances in our understanding of tumor immunology, which have in turn led to new and exciting immunology-based therapeutics. However, further research is needed into the dynamics and regulation of the immune response in the tumor microenvironment in order to achieve the full potential of these agents in treating all cancer patients. Defining the role of cytokines, chemokines, and other soluble mediators will be essential to this endeavor. This chapter describes, in detail, the technical protocol and applicability of LEGENDplex™ bead-based multiplex assays in quantifying these critical signaling molecules.


Asunto(s)
Biomarcadores de Tumor/análisis , Quimiocinas/análisis , Citometría de Flujo/métodos , Neoplasias/tratamiento farmacológico , Animales , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Biomarcadores de Tumor/inmunología , Biomarcadores de Tumor/metabolismo , Línea Celular Tumoral , Quimiocinas/inmunología , Quimiocinas/metabolismo , Monitoreo de Drogas/instrumentación , Monitoreo de Drogas/métodos , Citometría de Flujo/instrumentación , Humanos , Inmunoensayo/instrumentación , Inmunoensayo/métodos , Neoplasias/sangre , Neoplasias/inmunología , Neoplasias/patología , Microambiente Tumoral/efectos de los fármacos , Microambiente Tumoral/inmunología
15.
Scand J Pain ; 20(1): 125-138, 2019 12 18.
Artículo en Inglés | MEDLINE | ID: mdl-31584875

RESUMEN

It has been suggested that alterations in inflammation molecules maintain chronic pain although little is known about how these factors influence homeostatic and inflammatory events in common chronic pain conditions. Nonpharmacological interventions might be associated with alterations in inflammation markers in blood. This study of patients with chronic pain investigates whether an interdisciplinary multimodal rehabilitation program (IMMRP) was associated with significant alterations in the plasma pattern of 68 cytokines/chemokines 1 year after rehabilitation and whether such changes were associated with clinical changes. Blood samples and self-reports of pain, psychological distress, and physical activity of 25 complex chronic pain patients were collected pre-IMMRP and at 12-month follow-up. Analyses of inflammatory proteins (cytokines/chemokines/growth factors) were performed directly in plasma using the multiplex immunoassay technology Meso Scale Discovery. This explorative pilot study found that 12 substances, mainly pro-inflammatory, decreased after IMMRP. In two other relatively small IMMRP studies, four of these proinflammatory markers were also associated with decreases. The pattern of cytokines/chemokines pre-IMMRP was associated with changes in psychological distress but not with pain or physical activity. The present study cannot impute cause and effect. These results together with the results of the two previous IMMRP studies suggest that there is a need for larger and more strictly controlled studies of IMMRP with respect to inflammatory markers in blood. Such studies need to consider responders/non-responders, additional therapies, involved pain mechanisms and diagnoses. This and the two other studies open up for developing biologically measurable outcomes from plasma. Such biomarkers will be an important tool for further development of IMMRP and possibly other treatments for patients w ith chronic pain.


Asunto(s)
Proteínas Sanguíneas/análisis , Quimiocinas , Dolor Crónico/rehabilitación , Inflamación/sangre , Adulto , Biomarcadores/análisis , Biomarcadores/sangre , Quimiocinas/análisis , Quimiocinas/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto
16.
Oxid Med Cell Longev ; 2019: 5965721, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31396302

RESUMEN

Objective: The status of metabolites of the nitric oxide (NO) pathway in patients with chronic wounds in the course of cardiometabolic diseases is largely unknown. Yet arginine supplementation and citrulline supplementation as novel therapeutic modalities aimed at increasing NO are tested. Material and Methods: Targeted metabolomics approach (LC-MS/MS) was applied to determine the concentrations of L-arginine, L-citrulline, asymmetric and symmetric dimethylarginines (ADMA and SDMA), and arginine/ADMA and arginine/SDMA ratios as surrogate markers of NO and arginine availability in ulnar and femoral veins, representing systemic and local levels of metabolites, in patients with chronic wounds in the course of cardiometabolic diseases (n = 59) as compared to patients without chronic wounds but with similar cardiometabolic burden (n = 55) and healthy individuals (n = 88). Results: Patients with chronic wounds had significantly lower systemic L-citrulline and higher ADMA and SDMA concentrations and lower L-arginine/ADMA and L-arginine/SDMA as compared to healthy controls. The presence of chronic wounds in patients with cardiometabolic diseases was associated with decreased L-arginine but with increased L-citrulline, ADMA, and SDMA concentrations and decreased L-arginine/ADMA and L-arginine/SDMA. Serum obtained from the ulnar and femoral veins of patients with chronic wounds differed by L-arginine concentrations and L-arginine/SDMA ratio, both lower in the femoral vein. Wound etiology affected L-citrulline and SDMA concentrations, lower and higher, respectively, in patients with venous stasis, and the L-arginine/SDMA ratio-lower in venous stasis. The wound type affected L-arginine/ADMA and citrulline-lower in patients with ulcerations or gangrene. IL-6 was an independent predictor of L-arginine/ADMA, VEGF-A of ADMA, G-CSF of L-arginine/SDMA, and GM-CSF of L-citrulline and SDMA. Conclusion: Chronic wounds in the course of cardiometabolic diseases are associated with reduced NO and arginine availability due to ADMA and SDMA accumulation rather than arginine deficiency, not supporting its supplementation. Wound character seems to affect NO bioavailability and wound etiology-arginine bioavailability. Arginine concentration and its availability are more markedly reduced at the local level than the systemic level.


Asunto(s)
Arginina/metabolismo , Enfermedades Cardiovasculares/patología , Extremidad Inferior/patología , Óxido Nítrico/metabolismo , Heridas y Traumatismos/diagnóstico , Anciano , Arginina/análogos & derivados , Arginina/análisis , Enfermedades Cardiovasculares/complicaciones , Enfermedades Cardiovasculares/metabolismo , Estudios de Casos y Controles , Quimiocinas/análisis , Citrulina/análisis , Citocinas/análisis , Femenino , Hormona del Crecimiento/análisis , Humanos , Masculino , Metabolómica , Persona de Mediana Edad , Heridas y Traumatismos/complicaciones , Heridas y Traumatismos/metabolismo
17.
Int J Mol Sci ; 20(16)2019 Aug 09.
Artículo en Inglés | MEDLINE | ID: mdl-31404950

RESUMEN

Chemerin (CHEM) may act as an important link integrating energy homeostasis and reproductive functions of females, and its actions are mediated by three receptors: chemokine-like receptor 1 (CMKLR1), G protein-coupled receptor 1 (GPR1), and C-C motif chemokine receptor-like 2 (CCRL2). The aim of the current study was to compare the expression of CHEM and its receptor (CHEM system) mRNAs (quantitative real-time PCR) and proteins (Western blotting and fluorescent immunohistochemistry) in the selected areas of the porcine hypothalamus responsible for gonadotropin-releasing hormone production and secretion: the mediobasal hypothalamus, preoptic area and stalk median eminence during the oestrous cycle and early pregnancy. Moreover, plasma CHEM concentrations were determined using ELISA. The expression of CHEM system has been demonstrated in the porcine hypothalamus throughout the luteal phase and follicular phase of the oestrous cycle, and during early pregnancy from days 10 to 28. Plasma CHEM levels and concentrations of transcripts and proteins of CHEM system components in the hypothalamus fluctuated throughout pregnancy and the oestrous cycle. Our study was the first experiment to demonstrate the presence of CHEM system mRNAs and proteins in the porcine hypothalamus and the correlations between the expression levels and physiological hormonal milieu related to the oestrous cycle and early pregnancy.


Asunto(s)
Quimiocinas/análisis , Ciclo Estral , Hipotálamo/metabolismo , Receptores de Quimiocina/análisis , Animales , Quimiocinas/sangre , Quimiocinas/genética , Femenino , Expresión Génica , Hormona Liberadora de Gonadotropina/metabolismo , Hipotálamo/química , Embarazo , Receptores de Quimiocina/genética , Porcinos
18.
Methods Mol Biol ; 2024: 79-94, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31364043

RESUMEN

Multiplex bead array technology expands upon the principles of the enzyme-linked immunosorbent assay by allowing the simultaneous quantification of a large number of cytokines and chemokines within a single sample. This allows researchers more freedom and opportunities to investigate complex immune responses both in vivo and in vitro. Here we describe and update the detailed assay protocol and technical tips for simultaneous quantification of multiple cytokines and chemokines in mouse biological fluids such as sera, bronchoalveolar lavage fluid, tissue homogenate supernatant, and tissue culture supernatant, using a multiplex bead array assay.


Asunto(s)
Quimiocinas/análisis , Citocinas/análisis , Animales , Bioensayo/métodos , Líquido del Lavado Bronquioalveolar , Inmunoensayo/métodos , Ratones
19.
Exp Mol Med ; 51(7): 78, 2019 07 10.
Artículo en Inglés | MEDLINE | ID: mdl-31292433

RESUMEN

We hypothesized that NADPH oxidase 4 (Nox4) is involved in the formation of neointimal atherosclerotic plaques through the migration of smooth muscle cells (SMCs) in response to flagellin. Here, we demonstrate that TLR5-mediated Nox4 activation regulates the migration of SMCs, leading to neointimal plaque formation in atherosclerosis. To investigate the molecular mechanism by which the TLR5-Nox4 cascade mediates SMC migration, we analyzed the signaling cascade in primary vascular SMCs (VSMCs) from wild-type (WT) or Nox4 KO mice. Stimulation of VSMCs from Nox4 KO mice with flagellin failed to induce H2O2 production and Rac activation compared with stimulation of VSMCs from WT mice. Moreover, the migration of Nox4-deficient VSMCs was attenuated in response to flagellin in transwell migration and wound healing assays. Finally, we performed partial carotid artery ligation in ApoE KO and Nox4ApoE DKO mice fed a high-fat diet (HFD) with or without recombinant FliC (rFliC) injection. Injection of rFliC into ApoE KO mice fed a HFD resulted in significantly increased SMC migration into the intimal layer, whereas SMC accumulation was not detected in Nox4ApoE DKO mice. We conclude that activation of the TLR5-Nox4 cascade plays an important role in the formation of neointimal atherosclerotic plaques.


Asunto(s)
Aterosclerosis/enzimología , Flagelina/administración & dosificación , NADPH Oxidasa 4/metabolismo , Placa Aterosclerótica/enzimología , Transducción de Señal , Receptor Toll-Like 5/metabolismo , Animales , Aterosclerosis/patología , Arterias Carótidas/patología , Movimiento Celular , Quimiocinas/análisis , Dieta Alta en Grasa/efectos adversos , Flagelina/genética , Masculino , Ratones Noqueados para ApoE , Miocitos del Músculo Liso/enzimología , Miocitos del Músculo Liso/patología , NADPH Oxidasa 4/genética , Neointima/enzimología , Neointima/patología , Fenotipo , Placa Aterosclerótica/patología , Salmonella enteritidis/genética , Receptor Toll-Like 5/genética
20.
Balkan Med J ; 36(5): 270-275, 2019 08 22.
Artículo en Inglés | MEDLINE | ID: mdl-31284705

RESUMEN

Background: Chemerin is a recently discovered adipokine that plays a role in adipocyte metabolism. It is a novel chemoattractant adipokine whose expression and secretion are increased by adipogenesis. Aims: To evaluate the effects of probiotic supplementation on chemerin level, inflammation, and metabolic syndrome components in obese Wistar rats. Study Design: Animal experiment. Methods: We divided the experimental animals into three groups, each consisting of eight rats. Group 1 was the control group. Group 2 was the experimentally obese group, in which rats were fed with a high-fat diet. Group 3 was the obese intervention group, in which rats were supplemented with probiotics after obesity induction. Results: At the end of the study, a statistically significant difference was found between the groups in final weights, weight changes, and body mass index values (p<0.05). Weight gain was 34.12±3.70 g in group 3 post-probiotic supplementation and 53.25±8.35 g in group 2 (p<0.05). Obese rats showed increased levels of fasting plasma glucose, insulin, insulin resistance (homeostatic model assessment-insulin resistance), total cholesterol, low-density lipoprotein cholesterol, inflammatory markers, and leptin compared to those in the control group. Chemerin levels were 14.31±13.34 ng/mL in group 2 and 2.67±2.42 ng/mL in group 3 (p<0.05). Conclusion: Probiotic supplementation (group 3) reduced weight gain, and there were positive effects on the levels of fasting plasma glucose, insulin, homeostatic model assessment-insulin resistance, triglycerides, inflammatory markers, leptin, and chemerin.


Asunto(s)
Quimiocinas/análisis , Síndrome Metabólico/fisiopatología , Obesidad/fisiopatología , Probióticos/uso terapéutico , Animales , Quimiocinas/sangre , Modelos Animales de Enfermedad , Síndrome Metabólico/complicaciones , Síndrome Metabólico/patología , Obesidad/complicaciones , Obesidad/patología , Probióticos/farmacología , Ratas , Ratas Wistar , Estadísticas no Paramétricas
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