RESUMEN
Personalized therapies are designed to optimize the safety-to-efficacy ratio by selecting patients with higher response rates based on specific biomarkers. Inflammation plays a vital role in the pathogenesis of non-alcoholic steatohepatitis (NASH), a common liver disorder. Eotaxin-1 plays a role in innate and adaptive immune responses. High eotaxin-1 levels are associated with diabetes and fatty liver disease and, therefore, serves as a biomarker for patient selection. The anti-eotaxin-1 monoclonal antibody is tailored for the personalized therapy of patients with inflammatory conditions due to high levels of eotaxin-1. To evaluate the biological activity and immunomodulatory effect of orally administered anti-eotaxin-1. C57B1/6 mice were treated with either oral or intra-peritoneal anti-eotaxin-1 antibody before induction of immune-mediated hepatitis using an injection of concanavalin A (ConA) and checked for liver injury and eotaxin-1 serum levels. Oral administration of anti-eotaxin-1 alleviated the immune-mediated liver injury. Serum alanine aminotransferase levels decreased to 1807 U/L, compared with 19025 U/L in untreated controls and 3657 U/L in mice treated with parenteral anti-eotaxin-1 (P < 0.005). A trend toward reduced serum eotaxin-1 levels was observed in treated mice, ranging from 594 pg/mL in the controls to 554 and 561 pg/mL in mice treated orally and intraperitoneally (P = 0.08, P = 0.06, respectively). Oral administration of anti-eotaxin-1 antibody shows biological activity in the gut and exerts a systemic immunomodulatory effect to alleviate immune-mediated hepatitis. The data suggest that testing for eotaxin-1 serum levels may enable screening patients with high-eotaxin-1 levels-associated NASH.
Asunto(s)
Antiinflamatorios/uso terapéutico , Anticuerpos Monoclonales/uso terapéutico , Quimiocina CCL11/inmunología , Hepatitis Autoinmune/tratamiento farmacológico , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Medicina de Precisión/métodos , Administración Oral , Alanina Transaminasa/sangre , Animales , Anticuerpos Monoclonales/efectos adversos , Quimiocina CCL11/sangre , Concanavalina A , Hígado Graso/tratamiento farmacológico , Factores Inmunológicos/uso terapéutico , Inyecciones Intraperitoneales , Masculino , Ratones , Ratones Endogámicos C57BL , Enfermedad del Hígado Graso no Alcohólico/inducido químicamenteRESUMEN
Melanoma differentiation associated gene-9 (MDA-9), Syntenin-1, or syndecan binding protein is a differentially regulated prometastatic gene with elevated expression in advanced stages of melanoma. MDA-9/Syntenin expression positively associates with advanced disease stage in multiple histologically distinct cancers and negatively correlates with patient survival and response to chemotherapy. MDA-9/Syntenin is a highly conserved PDZ-domain scaffold protein, robustly expressed in a spectrum of diverse cancer cell lines and clinical samples. PDZ domains interact with a number of proteins, many of which are critical regulators of signaling cascades in cancer. Knockdown of MDA-9/Syntenin decreases cancer cell metastasis, sensitizing these cells to radiation. Genetic silencing of MDA-9/Syntenin or treatment with a pharmacological inhibitor of the PDZ1 domain, PDZ1i, also activates the immune system to kill cancer cells. Additionally, suppression of MDA-9/Syntenin deregulates myeloid-derived suppressor cell differentiation via the STAT3/interleukin (IL)-1ß pathway, which concomitantly promotes activation of cytotoxic T lymphocytes. Biologically, PDZ1i treatment decreases metastatic nodule formation in the lungs, resulting in significantly fewer invasive cancer cells. In summary, our observations indicate that MDA-9/Syntenin provides a direct therapeutic target for mitigating aggressive breast cancer and a small-molecule inhibitor, PDZ1i, provides a promising reagent for inhibiting advanced breast cancer pathogenesis.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Interleucina-1beta/genética , Neoplasias Pulmonares/tratamiento farmacológico , Oxadiazoles/farmacología , Pirimidinas/farmacología , Sinteninas/genética , Animales , Antineoplásicos/síntesis química , Neoplasias de la Mama/genética , Neoplasias de la Mama/inmunología , Neoplasias de la Mama/patología , Línea Celular Tumoral , Quimiocina CCL11/genética , Quimiocina CCL11/inmunología , Quimiocina CCL17/genética , Quimiocina CCL17/inmunología , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-1alfa/genética , Interleucina-1alfa/inmunología , Interleucina-1beta/antagonistas & inhibidores , Interleucina-1beta/inmunología , Subunidad p19 de la Interleucina-23/genética , Subunidad p19 de la Interleucina-23/inmunología , Interleucina-5/genética , Interleucina-5/inmunología , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/secundario , Ratones , Ratones Endogámicos BALB C , Oxadiazoles/síntesis química , Pirimidinas/síntesis química , Transducción de Señal , Sinteninas/antagonistas & inhibidores , Sinteninas/inmunología , Linfocitos T Citotóxicos/efectos de los fármacos , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/patología , Carga Tumoral/efectos de los fármacos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
AIM: We aimed to discover whether group 2 innate lymphoid cells (ILC2s) and cytokines in nasal lavage fluid could be used to predict eosinophilic infiltration in mice with eosinophilic chronic rhinosinusitis (ECRS). METHODS: Ten mice were divided into two groups. The ECRS group received an intranasal challenge of Aspergillus oryzae protease (AP) and ovalbumin (OVA) to establish disease. A control group received intranasal phosphate-buffered saline. Histopathology of nasal cavities and paranasal sinuses, and cytokine and ILC2s levels in nasal lavage fluid were analyzed and compared between the ECRS and control mouse groups. KEY FINDINGS: ILC2s numbers were not significantly higher in the nasal lavage fluid of the ECRS group mice compared with those of the control group. Eotaxin/chemokine (CC motif) ligand 11 (CCL11) levels were significantly higher in the nasal lavage fluid of mice in the ECRS group compared with those in the control group. However, no statistical differences were seen in the classic proinflammatory cytokines, IL-33, IL-25, and thymic stromal thymopoietin (TSLP), or the classic type 2 cytokines, IL-4, IL-5, and IL-13 between groups. SIGNIFICANCE: Eotaxin/CCL11 levels in nasal lavage fluid rather than that of ILC2s and classic proinflammatory and type 2 cytokines were significantly higher in ECRS mice compared with control ones. Eotaxin/CCL11 showed diagnostic and therapeutic value; however, more studies are needed to test and verify its value.
Asunto(s)
Quimiocina CCL11/metabolismo , Eosinofilia/metabolismo , Inmunidad Innata/fisiología , Líquido del Lavado Nasal , Sinusitis/metabolismo , Animales , Quimiocina CCL11/inmunología , Enfermedad Crónica , Eosinofilia/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Líquido del Lavado Nasal/inmunología , Valor Predictivo de las Pruebas , Sinusitis/inmunología , Sinusitis/patologíaRESUMEN
Particulate matter 2.5 (PM2.5) exposure can increase the prevalence of allergic rhinitis (AR), the mechanism underlying which may include oxidative stress and inflammatory response. As a ROS quenching agent, N-acetylcysteine (NAC) can attenuate the accumulation of inflammatory cells and hyper-responsiveness in animal asthma models. To explore the effect of NAC on the oxidative stress and inflammatory reactions in AR rats exposed to PM2.5, we analyzed the components of PM2.5 and examined the nasal symptoms, redox level in nasal mucosa, Th1/Th2-related serum cytokines, nasal mucosal histopathology and ultrastructure in AR rat models with NAC intervention after PM2.5 exposure. The results showed that the high concentrations of metal cations and PAHs in PM2.5 could aggravate Th2-dominant allergic inflammation in AR model and cause redox imbalance, accompanied by nasal epithelial cell stripping and eosinophil infiltration, while NAC intervention could alleviate the clinical symptoms of AR model after PM2.5 exposure, correct the redox imbalance, reduce the Th2 cytokines, reduce eosinophil infiltration, and promote the moderate regeneration of epithelial cells. The mechanism of NAC reversing PM2.5-mediated action may be related to its anti-oxidant and anti-inflammatory effects, which may provide some new insights for the prevention of AR exacerbated by exposure to PM2.5.
Asunto(s)
Acetilcisteína/farmacología , Antioxidantes/farmacología , Mucosa Nasal/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Rinitis Alérgica/tratamiento farmacológico , Balance Th1 - Th2/efectos de los fármacos , Animales , Quimiocina CCL11/genética , Quimiocina CCL11/inmunología , Modelos Animales de Enfermedad , Femenino , Expresión Génica , Inmunoglobulina E/genética , Inmunoglobulina E/inmunología , Inflamación , Interferón gamma/genética , Interferón gamma/inmunología , Interleucinas/genética , Interleucinas/inmunología , Malondialdehído/inmunología , Malondialdehído/metabolismo , Mucosa Nasal/inmunología , Mucosa Nasal/patología , Estrés Oxidativo/inmunología , Tamaño de la Partícula , Material Particulado/administración & dosificación , Hidrocarburos Policíclicos Aromáticos/administración & dosificación , Ratas , Ratas Sprague-Dawley , Rinitis Alérgica/inducido químicamente , Rinitis Alérgica/inmunología , Rinitis Alérgica/patología , Superóxido Dismutasa/genética , Superóxido Dismutasa/inmunologíaRESUMEN
Effective biomarkers for multiple sclerosis diagnosis, assessment of prognosis, and treatment responses, in particular those measurable in blood, are largely lacking. We have investigated a broad set of protein biomarkers in cerebrospinal fluid (CSF) and plasma using a highly sensitive proteomic immunoassay. Cases from two independent cohorts were compared with healthy controls and patients with other neurological diseases. We identified and replicated 10 cerebrospinal fluid proteins including IL-12B, CD5, MIP-1a, and CXCL9 which had a combined diagnostic efficacy similar to immunoglobulin G (IgG) index and neurofilament light chain (area under the curve [AUC] = 0.95). Two plasma proteins, OSM and HGF, were also associated with multiple sclerosis in comparison to healthy controls. Sensitivity and specificity of combined CSF and plasma markers for multiple sclerosis were 85.7% and 73.5%, respectively. In the discovery cohort, eotaxin-1 (CCL11) was associated with disease duration particularly in patients who had secondary progressive disease (PCSF < 4 × 10-5, Pplasma < 4 × 10-5), and plasma CCL20 was associated with disease severity (P = 4 × 10-5), although both require further validation. Treatment with natalizumab and fingolimod showed different compartmental changes in protein levels of CSF and peripheral blood, respectively, including many disease-associated markers (e.g., IL12B, CD5) showing potential application for both diagnosing disease and monitoring treatment efficacy. We report a number of multiple sclerosis biomarkers in CSF and plasma for early disease detection and potential indicators for disease activity. Of particular importance is the set of markers discovered in blood, where validated biomarkers are lacking.
Asunto(s)
Quimiocina CCL11/análisis , Quimiocina CCL20/sangre , Inflamación/inmunología , Esclerosis Múltiple/diagnóstico , Adulto , Biomarcadores/sangre , Biomarcadores/líquido cefalorraquídeo , Estudios de Casos y Controles , Quimiocina CCL11/inmunología , Quimiocina CCL20/inmunología , Estudios de Cohortes , Femenino , Humanos , Inflamación/sangre , Inflamación/líquido cefalorraquídeo , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/sangre , Esclerosis Múltiple/líquido cefalorraquídeo , Esclerosis Múltiple/inmunología , Pronóstico , Proteómica , Reproducibilidad de los Resultados , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
Although elevated blood or sputum eosinophils are present in many patients with COPD, uncertainties remain regarding the anatomical distribution pattern of lung-infiltrating eosinophils. Basophils have remained virtually unexplored in COPD. This study mapped tissue-infiltrating eosinophils, basophils and eosinophil-promoting immune mechanisms in COPD-affected lungs.Surgical lung tissue and biopsies from major anatomical compartments were obtained from COPD patients with severity grades Global Initiative for Chronic Obstructive Lung Disease stages I-IV; never-smokers/smokers served as controls. Automated immunohistochemistry and in situ hybridisation identified immune cells, the type 2 immunity marker GATA3 and eotaxins (CCL11, CCL24).Eosinophils and basophils were present in all anatomical compartments of COPD-affected lungs and increased significantly in very severe COPD. The eosinophilia was strikingly patchy, and focal eosinophil-rich microenvironments were spatially linked with GATA3+ cells, including type 2 helper T-cell lymphocytes and type 2 innate lymphoid cells. A similarly localised and interleukin-33/ST2-dependent eosinophilia was demonstrated in influenza-infected mice. Both mice and patients displayed spatially confined eotaxin signatures with CCL11+ fibroblasts and CCL24+ macrophages.In addition to identifying tissue basophilia as a novel feature of advanced COPD, the identification of spatially confined eosinophil-rich type 2 microenvironments represents a novel type of heterogeneity in the immunopathology of COPD that is likely to have implications for personalised treatment.
Asunto(s)
Basófilos/inmunología , Eosinófilos/inmunología , Macrófagos/inmunología , Enfermedad Pulmonar Obstructiva Crónica/inmunología , Eosinofilia Pulmonar/etiología , Adulto , Anciano , Animales , Biomarcadores , Quimiocina CCL11/inmunología , Quimiocina CCL24/inmunología , Femenino , Factor de Transcripción GATA3/inmunología , Humanos , Inmunidad Innata , Masculino , Ratones , Persona de Mediana Edad , Fumadores , Adulto JovenRESUMEN
BACKGROUND: Partial burn injury in older patients is associated with higher rates of morbidity, mortality, and conversion to full thickness burn (Finnerty et al., 2009; Pham et al., 2009). Both human and mouse models demonstrate an altered systemic immune response in older subjects, however less is known about the localized response (Jeschke et al., 2016; Farinas et al., 2018; Mohs et al., 2017). We hypothesized that a mouse model could demonstrate differences in the localized inflammatory response of the old. METHODS: Six old (66 weeks) and young (8 weeks) mice received partial thickness thermal burns. Localized and systemic expression of nine chemokines (TNFalpha, MCP-1, MIP-2, S100A9, EGF, IL-10, RANTES, G-CSF, and EOTAXIN) were evaluated at day 3 after burn using Luminex analysis. Vimentin immunostaining was used to evaluate injury depth. RESULTS: Vimentin staining demonstrated increased burn depth in old mice (449±38µm) as compared to young (166±18µm) (p<0.05). Both groups exhibited increased localized expression of EOTAXIN after burn (p<0.05), however expression in old mice (83.6±6.1pg/ml) was lower than that of young (126.8±18.7pg/ml) (p<0.05). Systemically, however, old mice had increased baseline EOTAXIN expression (1332.40±110.78pg/ml) compared to young (666.12±45.8pg/ml) (p<0.005). CONCLUSIONS: EOTAXIN is one of the primary chemoattractants for selective eosinophilic recruitment and activation. While eosinophils are important for wound healing, a hyperactive eosinophilic response can result in tissue damage. We hypothesize that the increased baseline serum EOTAXIN in the old may prime their hyperactive response, and may contribute to their worse clinical outcomes. Long-term eosinophil activation requires further study, however our findings indicate a role for EOTAXIN and eosinophils in burn response.
Asunto(s)
Envejecimiento/inmunología , Quemaduras/inmunología , Quimiocina CCL11/inmunología , Quimiocina CCL24/inmunología , Quimiocina CCL26/inmunología , Eosinófilos/inmunología , Envejecimiento/metabolismo , Animales , Quemaduras/metabolismo , Quemaduras/patología , Calgranulina B/inmunología , Calgranulina B/metabolismo , Quimiocina CCL11/metabolismo , Quimiocina CCL2/inmunología , Quimiocina CCL2/metabolismo , Quimiocina CCL24/metabolismo , Quimiocina CCL26/metabolismo , Quimiocina CCL5/inmunología , Quimiocina CCL5/metabolismo , Quimiocina CXCL2/inmunología , Quimiocina CXCL2/metabolismo , Eosinófilos/metabolismo , Factor de Crecimiento Epidérmico/inmunología , Factor de Crecimiento Epidérmico/metabolismo , Factor Estimulante de Colonias de Granulocitos/inmunología , Factor Estimulante de Colonias de Granulocitos/metabolismo , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Interleucina-10/inmunología , Interleucina-10/metabolismo , Ratones , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
High aspect ratio zinc oxide nanowires (ZnONWs) have become one of the most important products in nanotechnology. The wide range applications of ZnONWs have heightened the need for evaluating the risks and biological consequences to these particles. In this study, we investigated inflammatory pathways activated by ZnONWs in cultured cells as well as the consequences of systemic exposure in mouse models. Confocal microscopy showed rapid phagocytic uptake of FITC-ZnONWs by macrophages. Exposure of macrophages or lung epithelial cells to ZnONWs induced the production of CCL2 and CCL11. Moreover, ZnONWs exposure induced both IL-6 and TNF-α production only in macrophages but not in LKR13 cells. Intratracheal instillation of ZnONWs in C57BL/6 mice induced a significant increase in the total numbers of immune cells in the broncho alveolar lavage fluid (BALFs) 2 days after instillation. Macrophages and eosinophils were the predominant cellular infiltrates of ZnONWs exposed mouse lungs. Similar cellular infiltrates were also observed in a mouse air-pouch model. Pro-inflammatory cytokines IL-6 and TNF-α as well as chemokines CCL11, and CCL2 were increased both in BALFs and air-pouch lavage fluids. These results suggest that exposure to ZnONWs may induce distinct inflammatory responses through phagocytic uptake and formation of soluble Zn2+ ions.
Asunto(s)
Quimiocina CCL11/inmunología , Eosinófilos/efectos de los fármacos , Eosinófilos/inmunología , Inflamación/etiología , Nanocables/efectos adversos , Óxido de Zinc/efectos adversos , Animales , Quimiocina CCL11/genética , Quimiocina CCL2/genética , Modelos Animales de Enfermedad , Técnicas In Vitro , Inflamación/genética , Inflamación/inmunología , Mediadores de Inflamación/metabolismo , Interleucina-6/genética , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Ratones Endogámicos C57BL , Nanocables/química , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Células RAW 264.7 , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factor de Necrosis Tumoral alfa/genética , Regulación hacia Arriba/efectos de los fármacos , Óxido de Zinc/químicaRESUMEN
Eosinophils are the prominent inflammatory cell involved in allergic asthma, atopic dermatitis, eosinophilic esophagitis, and hypereosinophilic syndrome and are found in high numbers in local tissue and/or circulating blood of affected patients. There is recent interest in a family of alarmins, including TSLP, IL-25 and IL-33, that are epithelial-derived and released upon stimulation of epithelial cells. Several genome wide association studies have found SNPs in genes encoding IL-33 to be risk factors for asthma. In two studies examining the direct role of IL-33 in eosinophils, there were differences in eosinophil responses. We sought to further characterize activation of eosinophils with IL-33 compared to activation by other cytokines and chemokines. We assessed IL-33 stimulated adhesion, degranulation, chemotaxis and cell surface protein expression in comparison to IL-3, IL-5, and eotaxin-1 on human eosinophils. Our results demonstrate that IL-33 can produce as potent eosinophil activation as IL-3, IL-5 and eotaxin-1. Thus, when considering specific cytokine targeting strategies, IL-33 will be important to consider for modulating eosinophil function.
Asunto(s)
Asma/inmunología , Eosinófilos/inmunología , Interleucina-33/inmunología , Interleucina-5/inmunología , Rinitis Alérgica/inmunología , Adulto , Adhesión Celular , Células Cultivadas , Quimiocina CCL11/inmunología , Quimiocina CCL11/farmacología , Quimiotaxis , Eosinófilos/efectos de los fármacos , Femenino , Humanos , Interleucina-33/farmacología , Interleucina-5/farmacología , Masculino , Persona de Mediana EdadRESUMEN
Post-translational modification of chemokines mediated by the dipeptidyl peptidase DPP4 (CD26) has been shown to negatively regulate lymphocyte trafficking, and its inhibition enhances T cell migration and tumor immunity by preserving functional chemokine CXCL10. By extending those initial findings to pre-clinical models of hepatocellular carcinoma and breast cancer, we discovered a distinct mechanism by which inhibition of DPP4 improves anti-tumor responses. Administration of the DPP4 inhibitor sitagliptin resulted in higher concentrations of the chemokine CCL11 and increased migration of eosinophils into solid tumors. Enhanced tumor control was preserved in mice lacking lymphocytes and was ablated after depletion of eosinophils or treatment with degranulation inhibitors. We further demonstrated that tumor-cell expression of the alarmin IL-33 was necessary and sufficient for eosinophil-mediated anti-tumor responses and that this mechanism contributed to the efficacy of checkpoint-inhibitor therapy. These findings provide insight into IL-33- and eosinophil-mediated tumor control, revealed when endogenous mechanisms of DPP4 immunoregulation are inhibited.
Asunto(s)
Dipeptidil Peptidasa 4/inmunología , Eosinófilos/inmunología , Interleucina-33/inmunología , Neoplasias Experimentales/inmunología , Animales , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Movimiento Celular/inmunología , Quimiocina CCL11/inmunología , Quimiocina CCL11/metabolismo , Dipeptidil Peptidasa 4/metabolismo , Inhibidores de la Dipeptidil-Peptidasa IV/farmacología , Modelos Animales de Enfermedad , Eosinófilos/efectos de los fármacos , Eosinófilos/metabolismo , Humanos , Interleucina-33/metabolismo , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Neoplasias Experimentales/metabolismo , Neoplasias Experimentales/prevención & control , Fosfato de Sitagliptina/farmacologíaRESUMEN
Baseline eosinophils reside in the gastrointestinal tract; however, in several allergic disorders, excessive eosinophils accumulate in the blood as well in the tissues. Recently, we showed in vitro that interleukin (IL)-18 matures and transforms IL-5-generated eosinophils into the pathogenic eosinophils that are detected in human allergic diseases. To examine the role of local induction of IL-18 in promoting eosinophil-associated intestinal disorders, we generated enterocyte IL-18-overexpressing mice using the rat intestinal fatty acid-binding promoter (Fabpi) and analysed tissue IL-18 overexpression and eosinophilia by performing real-time polymerase chain reaction, Enzyme-Linked Immunosorbent Assay and anti-major basic protein immunostaining. Herein we show that Fabpi-IL-18 mice display highly induced IL-18 mRNA and protein in the jejunum. IL-18 overexpression in enterocytes promotes marked increases of eosinophils in the blood and jejunum. Our analysis shows IL-18 overexpression in the jejunum induces a specific population of CD101+ CD274+ tissue eosinophils. Additionally, we observed comparable tissue eosinophilia in IL-13-deficient-Fabpi-IL-18 mice, and reduced numbers of tissue eosinophils in eotaxin-deficient-Fabpi-IL-18 and IL-5-deficient-Fabpi-IL-18 mice compared with Fabpi-IL-18 transgenic mice. Notably, jejunum eosinophilia in IL-5-deficient-Fabpi-IL-18 mice is significantly induced compared with wild-type mice, which indicates the direct role of induced IL-18 in the tissue accumulation of eosinophils and mast cells. Furthermore, we also found that overexpression of IL-18 in the intestine promotes eosinophil-associated peanut-induced allergic responses in mice. Taken together, we provide direct in vivo evidence that induced expression of IL-18 in the enterocytes promotes eotaxin-1, IL-5 and IL-13 independent intestinal eosinophilia, which signifies the clinical relevance of induced IL-18 in eosinophil-associated gastrointestinal disorders (EGIDs) to food allergens.
Asunto(s)
Enterocitos/inmunología , Eosinófilos/inmunología , Interleucina-18/inmunología , Yeyuno/inmunología , Hipersensibilidad al Cacahuete/inmunología , Animales , Antígenos CD/genética , Antígenos CD/inmunología , Antígeno B7-H1/genética , Antígeno B7-H1/inmunología , Quimiocina CCL11/genética , Quimiocina CCL11/inmunología , Enterocitos/patología , Eosinófilos/patología , Humanos , Interleucina-13/genética , Interleucina-13/inmunología , Interleucina-18/genética , Interleucina-5/genética , Interleucina-5/inmunología , Yeyuno/patología , Mastocitos/inmunología , Mastocitos/patología , Ratones , Ratones Transgénicos , Hipersensibilidad al Cacahuete/genética , Hipersensibilidad al Cacahuete/patología , RatasRESUMEN
BACKGROUND: Eosinophil-associated RNases (EARs) are stored preformed in eosinophil cytoplasmic secretory granules and have a key role in eosinophil effector functions in host defence and inflammatory disorders. However, the secretion mechanisms of EARs are poorly understood. OBJECTIVE: Our study aimed to understand the involvement of cytoskeleton machinery in EAR secretion. METHODS: Fresh human and mouse eosinophils were stimulated with CCL11, and the secretion of enzymatically active EARs was detected using an RNase activity assay. The involvement of cytoskeletal elements or microtubules was probed using specific inhibitors. RESULTS: We found that dynamic polymerization of microtubules and cytoskeletal elements, such as Rho and Rac, is required for chemokine-mediated EAR secretion from human and mouse eosinophils. However, inhibition of ROCK (Rho-associated protein kinase) increased EAR secretion in human and mouse eosinophils even in the absence of chemokine stimulation, suggesting ROCK negatively regulates EAR secretion. CONCLUSIONS: Collectively, these data suggest a cytoskeleton-dependent mechanism of EAR secretion from eosinophils, findings that are pertinent to host defence, allergy and other eosinophil-associated diseases.
Asunto(s)
Proteína Catiónica del Eosinófilo/inmunología , Eosinófilos/inmunología , Proteínas de Unión al GTP rac/inmunología , Quinasas Asociadas a rho/inmunología , Animales , Quimiocina CCL11/genética , Quimiocina CCL11/inmunología , Proteína Catiónica del Eosinófilo/genética , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Proteínas de Unión al GTP rac/genética , Quinasas Asociadas a rho/genéticaRESUMEN
BACKGROUND: Several lines of evidence suggest effects of dietary fat on prostate cancer (PCa) development and progression. Targeting omega (ω)-3:ω6 fatty acids (FA) ratio could be beneficial against PCa by favorably modulating inflammation. Here, we studied the effects of ω3- and ω6-enriched diets on prostate tumor growth and inflammatory response in androgen-deprived and non-deprived conditions. METHODS: Immune-competent eugonadal and castrated C57BL/6 mice were injected with TRAMP-C2 prostate tumor cells and daily fed with ω3- or ω6-enriched diet. FA and cytokine profiles were measured in blood and tumors using gas chromatography and multiplex immunoassay, respectively. Immune cell infiltration in tumors was profiled by multicolor flow cytometry. RESULTS: ω3-enriched diet decreased prostate TRAMP-C2 tumor growth in immune-competent eugonadal and castrated mice. Cytokines associated with Th1 immune response (IL-12 [p70], IFN-γ, GM-CSF) and eosinophil recruitment (eotaxin-1, IL-5, and IL-13) were significantly elevated in tumors of ω3-fed mice. Using in vitro experiments, we confirmed ω3 FA-induced eotaxin-1 secretion by tumor cells and that eotaxin-1 secretion was regulated by androgens. Analysis of immune cell infiltrating tumors showed no major difference of immune cells' abundance between ω3- and ω6-enriched diets. CONCLUSIONS: ω3-enriched diet reduces prostate tumor growth independently of androgen levels. ω3 FA can inhibit tumor cell growth and induce a local anti-tumor inflammatory response. These findings warrant further examination of dietary ω3's potential to slow down the progression of androgen-sensitive and castrate-resistant PCa by modulating immune cell function in tumors.
Asunto(s)
Progresión de la Enfermedad , Ácidos Grasos Omega-3/administración & dosificación , Inmunidad Celular/inmunología , Orquiectomía , Neoplasias de la Próstata/dietoterapia , Neoplasias de la Próstata/inmunología , Animales , Quimiocina CCL11/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Orquiectomía/tendencias , Neoplasias de la Próstata/patología , Carga Tumoral/inmunología , Células Tumorales CultivadasRESUMEN
OBJECTIVE: Pulp regeneration by stem cell transplantation declines due to age-related reduction. We hypothesized that administration of a cytokine together with the cell transplantation may improve the stem cell niche microenvironment and promote regeneration. CCL11 is implicated as a factor in aging. This investigation was performed to investigate the changes in the quality of the regenerated pulp by administration of CCL11 antibody in the aged mice and elucidate the underlying mechanisms. MATERIALS AND METHODS: Mobilized dental pulp stem cell (MDPSC) transplants were characterized in an ectopic tooth root transplantation model in both the aged and young mice. The amount of regenerated pulp tissue was analyzed in the transplants with continuous administration of CCL11 antibody compared with those without the antibody administration. Blood CCL11 levels were assessed at the onset of the experiment. Furthermore, immunostaining of CD68 together with CD11c or CD206 for M1 and M2 macrophage, respectively, were performed. Each double-positive cell count of M1 and M2 macrophages and M1/M2 ratio in the transplants with administration were compared with those without administration both in the aged and young mice. RESULTS: The administration of CCL11 antibody enhanced pulp regeneration and significantly reduced the blood CCL11 level in the aged mice. As the number of M1 macrophages decreased, the M1/M2 ratio in the treated aged mouse was less than that in the untreated aged mouse. There was, however, significant difference between the treated aged mouse and the untreated young mouse. CONCLUSION: CCL11 antibody has the potential to enhance and stimulate pulp regeneration in the aged mice.
Asunto(s)
Envejecimiento , Anticuerpos Neutralizantes/administración & dosificación , Quimiocina CCL11/antagonistas & inhibidores , Pulpa Dental/citología , Trasplante de Células Madre , Células Madre/citología , Reimplante Dental , Animales , Diferenciación Celular , Quimiocina CCL11/inmunología , Pulpa Dental/inmunología , Pulpa Dental/metabolismo , Ratones , Ratones SCID , Regeneración , Células Madre/inmunología , Células Madre/metabolismo , PorcinosRESUMEN
BACKGROUND AND OBJETIVE: The pathogenesis of asthma is dependent on the balance between regulatory and effector T cells, which display differential expression of CD25 and CD26. Therefore, alteration of circulating levels of sCD25 and sCD26 during allergic asthma could be conditioned by changes in leukocyte phenotype. Objectives: To analyze expression of CD25 and CD26 on T lymphocytes and their soluble derivatives (sCD25, sCD26) during stable phases of moderate-severe allergic asthma. METHODS: Cross-sectional study with 2 adult cohorts of allergic asthmatics. Clinical, anthropometric, pulmonary, hematological, and biochemical parameters were measured. Phenotyping was performed with flow cytometry in both circulating and cultured leukocytes. Dipeptidyl peptidase 4 (DPP4) activity was assayed in culture supernatants. RESULTS: In vitro studies revealed upregulation of CD26 on human T lymphocytes upon activation, especially under TH17-favoring conditions, and a correlation with soluble DPP4 activity (rs=0.641; P<.001). CD26 expression on lymphocytes was higher in asthmatics, while serum sCD26 was lower in women and patients. The latter finding could be associated with an expanded CD25low/CD26low/CD127low subset of effector CD4+ T cells in allergic asthma, with no changes in Treg percentages. However, women showed an increased Teff/Treg ratio, which could explain their greater susceptibility to asthma. CONCLUSIONS: Allergic asthma causes an increment in CD25lowCD26low helper T cells detected in stable stages. These changes are mirrored in serum and should be considered in the light of the downmodulating role of CD26 in major chemokines related to the pathogenesis of asthma such as CCL11 (eotaxin), CCL5 (RANTES), and CXCL12a (SDF-1α).
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Asma/inmunología , Linfocitos T CD4-Positivos/inmunología , Dipeptidil Peptidasa 4/inmunología , Hipersensibilidad/inmunología , Linfocitos T Reguladores/inmunología , Células Th17/inmunología , Adulto , Quimiocina CCL11/inmunología , Quimiocina CCL5/inmunología , Quimiocina CXCL12/inmunología , Estudios Transversales , Regulación hacia Abajo/inmunología , Femenino , Citometría de Flujo/métodos , Humanos , Subunidad alfa del Receptor de Interleucina-2/inmunología , Masculino , Persona de Mediana Edad , Regulación hacia Arriba/inmunología , Adulto JovenRESUMEN
Chemokines are soluble molecules directing immune cell trafficking and homing, mediating inflammation, and initiating immune responses to infection. In horses, the analysis of chemokines has been limited by the lack of specific antibodies. We generated mAbs specific for the equine C-C motif chemokine ligands (CCL) CCL2 (MCP-1), CCL3 (MIP-1α), CCL5 (RANTES) and CCL11 (eotaxin) using hybridoma technology. Antibody specificity was confirmed by intracellular staining of Chinese Hamster Ovary cells transfected with expression vectors encoding for CCL2, CCL3, CCL5, or CCL11. Transfectants were stained with the anti-CCL mAbs. Flow cytometric analysis confirmed the specificity of the different mAbs for the respective chemokine. In addition, equine PBMC were stained after isolation, culture in medium, or stimulation with LPS, or PMA and ionomycin. CCL2 was detected in few cluster of differentiation (CD)14+ monocytes in PBMC stimulated with PMA and ionomycin for 2 h. CCL3 was produced by CD14+ monocytes after 4-6 h culture in medium. After stimulation with PMA and ionomycin for 12-24 h, CCL3 was also expressed in lymphocytes, mainly in CD4+ T cells. Stimulation with LPS reduced the percentage of CCL3+ monocytes in PBMC. CCL5 was detected in PBMC ex vivo in CD4+ and CD8+ T cells. Culture of PBMC for longer than 6 h or stimulation with PMA and ionomycin reduced the percentage of CCL5+ cells. CCL11 was produced by CD4+ T cells in PBMC after stimulation with PMA and ionomycin for 4-24 h. After LPS stimulation of PBMC, CCL2, CCL5, and CCL11 production were comparable to culture in medium alone. ELISAs for each of the four chemokines were developed using pairs of anti-equine CCL mAbs. Supernatants from PMA and ionomycin stimulated PBMC contained detectable amounts of CCL2, CCL3 and CCL5, while CCL11 secretion could be stimulated from equine tracheal epithelial cells in response to IL-4. The newly generated mAbs for equine CCL chemokines facilitate the quantitative analysis of intracellular chemokine production by flow cytometry and soluble chemokines by ELISA. The CCL mAbs are valuable tools to improve the evaluation of innate immune responses in horses.
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Quimiocina CCL11/metabolismo , Quimiocina CCL2/metabolismo , Quimiocina CCL3/metabolismo , Quimiocina CCL5/metabolismo , Leucocitos Mononucleares/metabolismo , Animales , Anticuerpos Monoclonales/inmunología , Células CHO , Quimiocina CCL11/inmunología , Quimiocina CCL2/inmunología , Quimiocina CCL3/inmunología , Quimiocina CCL5/inmunología , Cricetulus , Ensayo de Inmunoadsorción Enzimática/veterinaria , Citometría de Flujo/veterinaria , Caballos/inmunología , Leucocitos Mononucleares/inmunología , Ratones Endogámicos BALB C/inmunología , Monocitos/metabolismo , Linfocitos T/metabolismoRESUMEN
Eotaxins are C-C motif chemokines first identified as potent eosinophil chemoattractants. They facilitate eosinophil recruitment to sites of inflammation in response to parasitic infections as well as allergic and autoimmune diseases such as asthma, atopic dermatitis, and inflammatory bowel disease. The eotaxin family currently includes three members: eotaxin-1 (CCL11), eotaxin-2 (CCL24), and eotaxin-3 (CCL26). Despite having only ~30% sequence homology to one another, each was identified based on its ability to bind the chemokine receptor, CCR3. Beyond their role in innate immunity, recent studies have shown that CCL11 and related molecules may directly contribute to degenerative processes in the central nervous system (CNS). CCL11 levels increase in the plasma and cerebrospinal fluid of both mice and humans as part of normal aging. In mice, these increases are associated with declining neurogenesis and impaired cognition and memory. In humans, elevated plasma levels of CCL11 have been observed in Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, and secondary progressive multiple sclerosis when compared to age-matched, healthy controls. Since CCL11 is capable of crossing the blood-brain barrier of normal mice, it is plausible that eotaxins generated in the periphery may exert physiological and pathological actions in the CNS. Here, we briefly review known functions of eotaxin family members during innate immunity, and then focus on whether and how these molecules might participate in the progression of neurodegenerative diseases.
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Quimiocina CCL11/inmunología , Quimiocina CCL24/inmunología , Quimiocina CCL26/inmunología , Inmunidad Innata/inmunología , Enfermedades Neurodegenerativas/inmunología , Envejecimiento/inmunología , Animales , Quimiocina CCL11/sangre , Quimiocina CCL11/líquido cefalorraquídeo , Quimiocina CCL24/sangre , Quimiocina CCL24/líquido cefalorraquídeo , Quimiocina CCL26/sangre , Quimiocina CCL26/líquido cefalorraquídeo , Humanos , Enfermedades Neurodegenerativas/sangre , Enfermedades Neurodegenerativas/líquido cefalorraquídeo , Receptores CCR3/inmunología , Receptores CCR3/metabolismoRESUMEN
Chronic airway inflammation is recognized as an essential process in the pathogenesis of asthma. Cytokine profiles derived from immune and inflammation cells such as T-helper (Th) cells, eosinophilia and neutrophilia are not limited to the Th2 type in asthma. However, little is understood about associations between Th2-low inflammatory cytokine profiles and risk of asthma in adults. A case-control study of 910 adult asthma and 881 healthy controls was conducted. Inflammatory cytokines screening was undertaken by high-throughput protein microarray technology, and Th17-related inflammatory cytokines (IL17A, IL-9, adipsin and CCL11) were finally selected. Associations between these four cytokines and adult asthma risk were analyzed by multivariate logistic regression models. We observed that plasma IL-17A and IL-9 levels were significantly increased in asthmatics when compared with controls. However, the plasma expressions of adipsin and CCL11 in asthmatics were significantly lower than that in health controls. The adjusted ORs (95%CI) of association between IL-17A, IL-9, adipsin and CCL11 expressions and adult asthma were 3.08 (1.91, 4.97), 1.93 (1.41, 2.64), 10.02 (6.99, 14.37) and 3.29 (2.36, 4.59), respectively (all P trend < 0.0001). Our results suggested that elevated IL-17A and IL-9 expressions and decreased levels of adipsin and CCL11 were positively associated with adult asthma.
Asunto(s)
Asma/genética , Quimiocina CCL11/genética , Interleucina-17/genética , Interleucina-9/genética , Células Th17/inmunología , Adulto , Asma/inmunología , Asma/patología , Biomarcadores/metabolismo , Estudios de Casos y Controles , Quimiocina CCL11/inmunología , Factor D del Complemento/genética , Factor D del Complemento/inmunología , Femenino , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Interleucina-17/inmunología , Interleucina-9/inmunología , Modelos Logísticos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Análisis de Secuencia por Matrices de Oligonucleótidos , Factores de Riesgo , Células Th17/patología , Células Th2/inmunología , Células Th2/patologíaRESUMEN
OBJECTIVE: To observe the effect of acupoint injection on eosinophils (EOS) counts and expression of eotaxin in nasal mucosa of allergic rhinitis (AR) rats, so as to reveal its mechanism underlying improving AR. METHODS: Twenty-four Sprague-Dawley (SD) rats were randomly divided into normal, model and acupoint injection groups (n=8 in each group). The AR model was established by intraperitoneal injection of ovalbumin sensitization. Bilateral "Yingxiang"(LI 20) and "Yintang"(GV 29) were selected for acupoint injection of the mixture solution of lidocaine, dexamethasone, and transfer factor (0.1 mL/acupoint) on the 1st, 5th, 9th, and 13th day after AR model established, a total of four times. EOS in the nasal mucosa was counted under light microscope after HE staining. Protein and mRNA expressions of eotaxin in the nasal mucosa were detected by immunohistochemical and RT-PCR methods, respectively. RESULTS: Compared with the normal group, EOS counts, protein and mRNA expressions of eotaxin in the nasal mucosa were significantly higher in the model group (P<0.05). Compared with the model group, EOS counts, protein and mRNA expressions of eotaxin in the nasal mucosa were significantly lower in the acupoint injection group (P<0.05). CONCLUSIONS: Acupoint injection can reduce the nasal mucosa inflammation by suppressing the protein and mRNA expressions of eotaxin, decreasing the infiltration and gathering of EOS in the nasal mucosa.
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Puntos de Acupuntura , Quimiocina CCL11/genética , Dexametasona/administración & dosificación , Eosinófilos/citología , Lidocaína/administración & dosificación , Rinitis Alérgica/tratamiento farmacológico , Factor de Transferencia/administración & dosificación , Animales , Quimiocina CCL11/inmunología , Eosinófilos/efectos de los fármacos , Eosinófilos/inmunología , Femenino , Humanos , Inyecciones , Recuento de Leucocitos , Masculino , Mucosa Nasal/efectos de los fármacos , Mucosa Nasal/inmunología , Ratas , Ratas Sprague-Dawley , Rinitis Alérgica/genética , Rinitis Alérgica/inmunologíaRESUMEN
In a transgenic mouse line hK14mIL33tg, with the expression of interleukin-33 (IL-33) driven by a keratin 14 promoter, keratoconjunctivitis developed spontaneously between 18 and 22 weeks of age under specific-pathogen-free conditions. These mice showed blepharitis and corneal impairments, and the histology revealed epithelial thickening in the conjunctiva and the cornea with infiltration of eosinophils, mast cells and basophils. IL-5, IL-13 and CCL11 were abundant in lacrimal fluid in the mice, and the gene expressions of IL-4, IL-5, IL-13, IL-33, Prg2 and Mmcp8 were significantly increased in the cornea. Furthermore, group 2 innate lymphoid cells (ILC2) producing IL-5 and IL-13 were markedly increased in the cornea. These phenotypes closely resemble human atopic keratoconjunctivitis (AKC). The characteristic ocular phenotype in these mice strongly suggests that IL-33 is crucial for the development of AKC. The mouse line may be useful as a novel model for research and development of therapeutic strategies for AKC.