RESUMEN
The severity of allergic asthma is driven by the balance between allergen-specific T regulatory (Treg) and T helper (Th)2 cells. However, it is unclear whether specific subsets of conventional dendritic cells (cDCs) promote the differentiation of these two T cell lineaeges. We have identified a subset of lung resident type 2 cDCs (cDC2s) that display high levels of CD301b and have potent Treg-inducing activity ex vivo. Single cell RNA sequencing and adoptive transfer experiments show that during allergic sensitization, many CD301b+ cDC2s transition in a stepwise manner to CD200+ cDC2s that selectively promote Th2 differentiation. GM-CSF augments the development and maintenance of CD301b+ cDC2s in vivo, and also selectively expands Treg-inducing CD301b+ cDC2s derived from bone marrow. Upon their adoptive transfer to recipient mice, lung-derived CD301b+ cDC2s confer immunological tolerance to inhaled allergens. Thus, GM-CSF maintains lung homeostasis by increasing numbers of Treg-inducing CD301b+ cDC2s.
RESUMEN
BACKGROUND: Allergic asthma is driven largely by allergen-specific TH2 cells, which develop in regional lymph nodes on the interaction of naive CD4+ T cells with allergen-bearing dendritic cells that migrate from the lung. This migration event is dependent on CCR7 and its chemokine ligand, CCL21. However, is has been unclear whether the other CCR7 ligand, CCL19, has a role in allergic airway disease. OBJECTIVE: This study sought to define the role of CCL19 in TH2 differentiation and allergic airway disease. METHODS: Ccl19-deficient mice were studied in an animal model of allergic asthma. Dendritic cells or fibroblastic reticular cells from wild-type and Ccl19-deficient mice were cultured with naive CD4+ T cells, and cytokine production was measured by ELISA. Recombinant CCL19 was added to CD4+ T-cell cultures, and gene expression was assessed by RNA-sequencing and quantitative PCR. Transcription factor activation was assessed by flow cytometry. RESULTS: Lungs of Ccl19-deficient mice had less allergic airway inflammation, reduced airway hyperresponsiveness, and less IL-4 and IL-13 production compared with lungs of Ccl19-sufficient animals. Naive CD4+ T cells cocultured with Ccl19-deficient dendritic cells or fibroblastic reticular cells produced lower amounts of type 2 cytokines than did T cells cocultured with their wild-type counterparts. Recombinant CCL19 increased phosphorylation of STAT5 and induced expression of genes associated with TH2 cell and IL-2 signaling pathways. CONCLUSIONS: These results reveal a novel, TH2 cell-inducing function of CCL19 in allergic airway disease and suggest that strategies to block this pathway might help to reduce the incidence or severity of allergic asthma.
Asunto(s)
Asma , Hipersensibilidad , Animales , Ratones , Quimiocina CCL19/genética , Receptores CCR7 , Ligandos , Asma/genética , Inflamación/patología , Pulmón , Hipersensibilidad/metabolismo , Alérgenos/metabolismo , Diferenciación Celular , Células Th2 , Células DendríticasRESUMEN
With only four classes of antifungal drugs available for the treatment of invasive systemic fungal infections, the number of resistant fungi is increasing, highlighting the urgent need for novel antifungal drugs. Ergosterol, an essential component of cell membranes, and its synthetic pathway have been targeted for antifungal drug development. Sterol-C4-methyl monooxygenase (Erg25p), which is a greater essential target than that of existing drugs, represents a promising drug target. However, the development of antifungal drugs must consider potential side effects, emphasizing the importance of evaluating their selective toxicity against fungi. In this study, we knocked in ERG25 of Candida glabrata and its human ortholog, SC4MOL, in ERG25-deleted Saccharomyces cerevisiae. Utilizing these strains, we evaluated 1181-0519, an Erg25p inhibitor, that exhibited selective toxicity against the C. glabrata ERG25 knock-in strain. Furthermore, 1181-0519 demonstrated broad-spectrum antifungal activity against pathogenic Candida species, including Candida auris. The approach of utilizing a gene that is functionally conserved between yeast and humans and subsequently screening for molecular target drugs enables the identification of selective inhibitors for both species.
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DNASE1L3, an enzyme highly expressed in DCs, is functionally important for regulating autoimmune responses to self-DNA and chromatin. Deficiency of DNASE1L3 leads to development of autoimmune diseases in both humans and mice. However, despite the well-established causal relationship between DNASE1L3 and immunity, little is known about the involvement of DNASE1L3 in regulation of antitumor immunity, the foundation of modern antitumor immunotherapy. In this study, we identify DNASE1L3 as a potentially new regulator of antitumor immunity and a tumor suppressor in colon cancer. In humans, DNASE1L3 is downregulated in tumor-infiltrating DCs, and this downregulation is associated with poor patient prognosis and reduced tumor immune cell infiltration in many cancer types. In mice, Dnase1l3 deficiency in the tumor microenvironment enhances tumor formation and growth in several colon cancer models. Notably, the increased tumor formation and growth in Dnase1l3-deficient mice are associated with impaired antitumor immunity, as evidenced by a substantial reduction of cytotoxic T cells and a unique subset of DCs. Consistently, Dnase1l3-deficient DCs directly modulate cytotoxic T cells in vitro. To our knowledge, our study unveils a previously unknown link between DNASE1L3 and antitumor immunity and further suggests that restoration of DNASE1L3 activity may represent a potential therapeutic approach for anticancer therapy.
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Neoplasias del Colon , Humanos , Ratones , Animales , Neoplasias del Colon/metabolismo , Cromatina/metabolismo , Inmunoterapia , Linfocitos T Citotóxicos , Microambiente Tumoral , Endodesoxirribonucleasas/genética , Endodesoxirribonucleasas/metabolismoRESUMEN
Nausea and vomiting in pregnancy (NVP) is a common symptom. Although the influence of NVP during the first trimester on dietary intake and birth outcomes has been revealed, no study has focused on NVP during the second trimester. This study aimed to reveal whether NVP severity during the second trimester is associated with dietary intake, gestational weight gain (GWG), birth weight, and delivery week. Participants completed a questionnaire at 18-27 gestational weeks. NVP severity was assessed using the modified Pregnancy-Unique Quantification of Emesis and Nausea scale in the questionnaire. Dietary habits were assessed using a brief-type diet history questionnaire. In total, 825 responses were analyzed: 202 (24.5%), 135 (16.4%), and 8 (1.0%) women reported mild, moderate, and severe NVP, respectively; 480 (58.2%) women did not have NVP during the second trimester. No significant association was observed between energy and nutrient intake and no/mild and moderate/severe NVP. Women with moderate/severe NVP had lower total GWG than those with no/mild NVP (p = 0.007). There was no significant difference in low birth weight and preterm birth rates (p = 0.246 and p = 0.604). This is the first study to investigate whether NVP severity during the second trimester is associated with dietary intake and birth outcomes.
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Complicaciones del Embarazo , Nacimiento Prematuro , Embarazo , Humanos , Recién Nacido , Femenino , Masculino , Segundo Trimestre del Embarazo , Estudios Prospectivos , Japón , Náusea/etiología , Vómitos , Ingestión de AlimentosRESUMEN
Mitochondrial dysfunction or morphological abnormalities in human pathogenic fungi are known to contribute to azole resistance; however, the underlying molecular mechanisms are unknown. In this study, we investigated the link between mitochondrial morphology and azole resistance in Candida glabrata, which is the second most common cause of human candidiasis worldwide. The ER-mitochondrial encounter structure (ERMES) complex is thought to play an important role in the mitochondrial dynamics necessary for mitochondria to maintain their function. Of the five components of the ERMES complex, deletion of GEM1 increased azole resistance. Gem1 is a GTPase that regulates the ERMES complex activity. Point mutations in GEM1 GTPase domains were sufficient to confer azole resistance. The cells lacking GEM1 displayed abnormalities in mitochondrial morphology, increased mtROS levels, and increased expression of azole drug efflux pumps encoded by CDR1 and CDR2. Interestingly, treatment with N-acetylcysteine (NAC), an antioxidant, reduced ROS production and the expression of CDR1 in Δgem1 cells. Altogether, the absence of Gem1 activity caused an increase in mitochondrial ROS concentration, leading to Pdr1-dependent upregulation of the drug efflux pump Cdr1, resulting in azole resistance.
RESUMEN
Seawater reverse osmosis (SWRO) is the most energy-efficient process for desalination to produce drinking water from seawater. However, its sustainability is still challenged by membrane fouling. Appropriate feed water quality is one of the crucial prerequisites for SWRO operation. In the current study, a ceramic adsorption filter (CAF), which was predominantly coated with an aluminum-based adsorbent (i.e., Alumina, Al2O3), was employed to enhance the pretreatment performance of SWRO. The fouling performance of SWRO pre-treated with a CAF was evaluated by feeding with real ultrafiltration (UF)-filtrated seawater collected from a seawater desalination R&D facility in Singapore. The flux decline profile showed that the presence of CAF after UF could mitigate around 10-30% of SWRO fouling. Based on the autopsy of the fouled SWRO membranes, it was observed that SWRO with CAF pre-treatment and daily regeneration could alleviate around 77.5% of Ca-induced inorganic fouling as well as 76% of lower biofouling. The present work highlights the potential of applying adsorption technology to enhance pre-treatment performance to extend the lifespan of SWRO membranes. Coupling the adsorbents on a ceramic filter should be a useful way to ease their implementation, i.e., inline adsorption and re-generation.
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The intensity and longevity of inflammatory responses to inhaled allergens is determined largely by the balance between effector and regulatory immune responses, but the mechanisms that determine the relative magnitudes of these opposing forces remain poorly understood. We have found that the type of adjuvant used during allergic sensitization has a profound effect on both the nature and longevity of the pulmonary inflammation triggered by subsequent reexposure to that same provoking allergen. TLR ligand adjuvants and house dust extracts primed immune responses characterized by a mixed neutrophilic and eosinophilic inflammation that was suppressed by multiple daily allergen challenges. During TLR ligand-mediated allergic sensitization, mice displayed transient airway neutrophilia, which triggered the release of TGF-ß into the airway. This neutrophil-dependent production of TGF-ß during sensitization had a delayed, suppressive effect on eosinophilic responses to subsequent allergen challenge. Neutrophil depletion during sensitization did not affect numbers of Foxp3+ Tregs but increased proportions of Gata3+CD4+ T cells, which, upon their transfer to recipient mice, triggered stronger eosinophilic inflammation. Thus, a neutrophil/TGF-ß axis acts during TLR-mediated allergic sensitization to fine-tune the phenotype of developing allergen-specific CD4+ T cells and limit their pathogenicity, suggesting a novel immunotherapeutic approach to control eosinophilia in asthma.
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Alérgenos/inmunología , Linfocitos T CD4-Positivos/inmunología , Neutrófilos/metabolismo , Hipersensibilidad Respiratoria/inmunología , Células Th2/inmunología , Factor de Crecimiento Transformador beta/inmunología , Animales , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Linfocitos T CD4-Positivos/patología , Modelos Animales de Enfermedad , Recuento de Linfocitos , Masculino , Ratones , Ratones Endogámicos C57BL , Neutrófilos/patología , Hipersensibilidad Respiratoria/patología , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Dendritic cells (DC) in the lung that induce Th17 differentiation remain incompletely understood, in part because conventional CD11b+ DCs (cDC2) are heterogeneous. Here, we report a population of cDCs that rapidly accumulates in lungs of mice following house dust extract inhalation. These cells are Ly-6C+, are developmentally and phenotypically similar to cDC2, and strongly promote Th17 differentiation ex vivo. Single cell RNA-sequencing (scRNA-Seq) of lung cDC2 indicates 5 distinct clusters. Pseudotime analysis of scRNA-Seq data and adoptive transfer experiments with purified cDC2 subpopulations suggest stepwise developmental progression of immature Ly-6C+Ly-6A/E+ cDC2 to mature Ly-6C-CD301b+ lung resident cDC2 lacking Ccr7 expression, which then further mature into CD200+ migratory cDC2 expressing Ccr7. Partially mature Ly-6C+Ly-6A/E-CD301b- cDC2, which express Il1b, promote Th17 differentiation. By contrast, CD200+ mature cDC2 strongly induce Th2, but not Th17, differentiation. Thus, Th17 and Th2 differentiation are promoted by lung cDC2 at distinct stages of maturation.
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Asma/inmunología , Antígeno CD11b/inmunología , Células Dendríticas/inmunología , Pulmón/inmunología , Células Th17/inmunología , Células Th2/inmunología , Traslado Adoptivo/métodos , Animales , Asma/metabolismo , Asma/patología , Secuencia de Bases , Antígeno CD11b/metabolismo , Diferenciación Celular/fisiología , Células Cultivadas , Técnicas de Cocultivo , Células Dendríticas/citología , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Pulmón/metabolismo , Pulmón/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Análisis de la Célula Individual/métodos , Células Th17/citología , Células Th2/citologíaRESUMEN
Airway eosinophilia is a hallmark of allergic asthma and is associated with mucus production, airway hyperresponsiveness, and shortness of breath. Although glucocorticoids are widely used to treat asthma, their prolonged use is associated with several side effects. Furthermore, many individuals with eosinophilic asthma are resistant to glucocorticoid treatment, and they have an unmet need for novel therapies. Here, we show that UDP-glucose (UDP-G), a nucleotide sugar, is selectively released into the airways of allergen-sensitized mice upon their subsequent challenge with that same allergen. Mice lacking P2Y14R, the receptor for UDP-G, had decreased airway eosinophilia and airway hyperresponsiveness compared with wild-type mice in a protease-mediated model of asthma. P2Y14R was dispensable for allergic sensitization and for the production of type 2 cytokines in the lung after challenge. However, UDP-G increased chemokinesis in eosinophils and enhanced their response to the eosinophil chemoattractant, CCL24. In turn, eosinophils triggered the release of UDP-G into the airway, thereby amplifying eosinophilic recruitment. This positive feedback loop was sensitive to therapeutic intervention, as a small molecule antagonist of P2Y14R inhibited airway eosinophilia. These findings thus reveal a pathway that can be therapeutically targeted to treat asthma exacerbations and glucocorticoid-resistant forms of this disease.
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Asma/inmunología , Eosinófilos/inmunología , Eosinofilia Pulmonar/inmunología , Receptores Purinérgicos P2Y/inmunología , Uridina Difosfato Glucosa/inmunología , Alérgenos/inmunología , Animales , Asma/genética , Asma/patología , Quimiocina CCL24/genética , Quimiocina CCL24/inmunología , Eosinófilos/patología , Masculino , Ratones , Ratones Noqueados , Eosinofilia Pulmonar/genética , Eosinofilia Pulmonar/patología , Receptores Purinérgicos P2Y/deficiencia , Células Th2/inmunología , Células Th2/patología , Uridina Difosfato Glucosa/genéticaRESUMEN
Airway neutrophilia occurs in approximately 50% of patients with asthma and is associated with particularly severe disease. Unfortunately, this form of asthma is usually refractory to corticosteroid treatment, and there is an unmet need for new therapies. Pulmonary neutrophilic inflammation is associated with Th17 cells, whose differentiation is controlled by the nuclear receptor, RORγt. Here, we tested whether VTP-938, a selective inverse agonist of this receptor, can reduce disease parameters in animal models of neutrophilic asthma. When administered prior to allergic sensitization through the airway, the RORγt inverse agonist blunted allergen-specific Th17 cell development in lung-draining lymph nodes and attenuated allergen-induced production of IL-17. VTP-938 also reduced pulmonary production of IL-17 and airway neutrophilia when given during the allergen challenge of the model. Finally, in an environmentally relevant model of allergic responses to house dust extracts, VTP-938 suppressed production of IL-17 and neutrophilic inflammation, and also markedly diminished airway hyperresponsiveness. Together, these findings suggest that orally available inverse agonists of RORγt might provide an effective therapy to treat glucocorticoid-resistant neutrophilic asthma.
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Asma/tratamiento farmacológico , Hipersensibilidad/tratamiento farmacológico , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/uso terapéutico , Hipersensibilidad Respiratoria/tratamiento farmacológico , Animales , Modelos Animales de Enfermedad , Polvo , Hipersensibilidad/inmunología , Inflamación , Interleucina-17 , Pulmón/patología , Ratones , Ratones Endogámicos C57BL , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/inmunología , Neumonía , Células Th17/inmunologíaRESUMEN
It is well established that responses to inhaled environmental agents are controlled by the coordinated actions by multiple immune cell types, including macrophages, dendritic cells, and lymphocytes. Recent evidence indicates that some structural cells can also contribute to the initiation and propagation of immune responses. For example, airway epithelial cells can promote eosinophilic inflammation in response to allergen inhalation. Much remains to be learned, however, regarding how each of these cell types interact with the others, and how these interactions shape immune responses to inhaled agents. Such studies have been hampered by the lack of reliable methods to isolate multiple and distinct populations of cells from the same tissue sample. Consequently, investigators have had to choose between using different protocols to isolate different populations of cells from different animals and accept that for some populations, cell yields can be very low. To overcome these difficulties, we have developed a convenient and practical method to isolate and purify subpopulations of epithelial and endothelial cells from mouse lung. Here, we describe these methods in detail.
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Células Epiteliales Alveolares/citología , Células Epiteliales Alveolares/metabolismo , Separación Celular , Células Endoteliales/citología , Células Endoteliales/metabolismo , Pulmón/citología , Animales , Biomarcadores , Separación Celular/métodos , Citometría de Flujo/métodos , Inmunohistoquímica , Pulmón/inmunología , Ratones , Células del Estroma/metabolismoRESUMEN
Asthma is associated with exposure to a wide variety of allergens and adjuvants. The extent to which overlap exists between the cellular and molecular mechanisms triggered by these various agents is poorly understood, but it might explain the differential responsiveness of patients to specific therapies. In particular, it is unclear why some, but not all, patients benefit from blockade of TNF. Here, we characterized signaling pathways triggered by distinct types of adjuvants during allergic sensitization. Mice sensitized to an innocuous protein using TLR ligands or house dust extracts as adjuvants developed mixed eosinophilic and neutrophilic airway inflammation and airway hyperresponsiveness (AHR) following allergen challenge, whereas mice sensitized using proteases as adjuvants developed predominantly eosinophilic inflammation and AHR. TLR ligands, but not proteases, induced TNF during allergic sensitization. TNF signaled through airway epithelial cells to reprogram them and promote Th2, but not Th17, development in lymph nodes. TNF was also required during the allergen challenge phase for neutrophilic and eosinophilic inflammation. In contrast, TNF was dispensable for allergic airway disease in a protease-mediated model of asthma. These findings might help to explain why TNF blockade improves lung function in only some patients with asthma.
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Hipersensibilidad/metabolismo , Inflamación/fisiopatología , Hipersensibilidad Respiratoria/metabolismo , Receptores Toll-Like/metabolismo , Factor de Necrosis Tumoral alfa/fisiología , Alérgenos , Animales , Asma/metabolismo , Asma/fisiopatología , Hiperreactividad Bronquial/fisiopatología , Diferenciación Celular , Citocinas/metabolismo , Modelos Animales de Enfermedad , Eosinófilos/citología , Hipersensibilidad/fisiopatología , Interleucina-17/metabolismo , Ligandos , Pulmón/fisiopatología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/citología , Ovalbúmina/metabolismo , Hipersensibilidad Respiratoria/fisiopatología , Transducción de Señal , Células Th17/citología , Células Th2/citologíaRESUMEN
Precursors of dendritic cells (pre-DCs) arise in the bone marrow (BM), egress to the blood, and finally migrate to peripheral tissue, where they differentiate to conventional dendritic cells (cDCs). Upon their activation, antigen-bearing cDCs migrate from peripheral tissue to regional lymph nodes (LNs) in a manner dependent on the chemokine receptor, CCR7. To maintain immune homeostasis, these departing cDCs must be replenished by new cDCs that develop from pre-DCs, but the molecular signals that direct pre-DC trafficking from the BM to the blood and peripheral tissues remain poorly understood. In the present study, we found that pre-DCs express the chemokine receptors CXCR4, CCR2, and CX3CR1, and that each of these receptors has a distinct role in pre-DC trafficking. Flow cytometric analysis of pre-DCs lacking CXCR4 revealed that this receptor is required for the retention of pre-DCs in the BM. Analyses of mice lacking CCR2 or CX3CR1, or both, revealed that they promote pre-DC migration to the lung at steady state. CCR2, but not CX3CR1, was required for pre-DC migration to the inflamed lung. Thus, these multiple chemokine receptors cooperate in a step-wise fashion to coordinate the trafficking of pre-DCs from the BM to the circulation and peripheral tissues.
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Células de la Médula Ósea/inmunología , Células Dendríticas/inmunología , Pulmón/inmunología , Neumonía/inmunología , Receptores CCR2/inmunología , Receptores CXCR4/inmunología , Receptores de Quimiocina/inmunología , Animales , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/patología , Receptor 1 de Quimiocinas CX3C , Diferenciación Celular , Movimiento Celular/efectos de los fármacos , Células Dendríticas/efectos de los fármacos , Células Dendríticas/patología , Regulación de la Expresión Génica , Lipopolisacáridos , Pulmón/efectos de los fármacos , Pulmón/patología , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neumonía/inducido químicamente , Neumonía/genética , Neumonía/patología , Receptores CCR2/deficiencia , Receptores CCR2/genética , Receptores CCR7/genética , Receptores CCR7/inmunología , Receptores CXCR4/deficiencia , Receptores CXCR4/genética , Receptores de Quimiocina/deficiencia , Receptores de Quimiocina/genética , Transducción de SeñalRESUMEN
The induction of allergen-specific T helper 2 (Th2) cells by lung dendritic cells (DCs) is a critical step in allergic asthma development. Airway delivery of purified allergens or microbial products can promote Th2 priming by lung DCs, but how environmentally relevant quantities and combinations of these factors affect lung DC function is unclear. Here, we investigated the ability of house dust extract (HDE), which contains a mixture of environmental adjuvants, to prime Th2 responses against an innocuous inhaled antigen. Inhalational exposure to HDE conditioned lung conventional DCs, but not monocyte-derived DCs, to induce antigen-specific Th2 differentiation. Conditioning of DCs by HDE was independent of Toll-like receptor 4 signaling, indicating that environmental endotoxin is dispensable for programming DCs to induce Th2 responses. DCs directly treated with HDE underwent maturation but were poor stimulators of Th2 differentiation. In contrast, DCs treated with bronchoalveolar lavage fluid (BALF) from HDE-exposed mice induced robust Th2 differentiation. DC conditioning by BALF was independent of the proallergic cytokines IL-25, IL-33, and thymic stromal lymphopoietin. BALF treatment of DCs resulted in upregulation of CD80 but low expression of CD40, CD86, and IL-12p40, which was associated with Th2 induction. These findings support a model whereby environmental adjuvants in house dust indirectly program DCs to prime Th2 responses by triggering the release of endogenous soluble factor(s) by airway cells. Identifying these factors could lead to novel therapeutic targets for allergic asthma.
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Células Dendríticas/inmunología , Polvo/inmunología , Pulmón/patología , Células Th2/inmunología , Animales , Antígenos CD/metabolismo , Asma/inmunología , Asma/metabolismo , Células Cultivadas , Técnicas de Cocultivo , Interleucinas/metabolismo , Pulmón/inmunología , Masculino , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
Differential display of the integrins CD103 and CD11b are widely used to distinguish two major dendritic cell (DC) subsets in nonlymphoid tissues. CD103(+) DCs arise from FLT3-dependent DC precursors (preDCs), whereas CD11b(hi) DCs can arise either from preDCs or FLT3-independent monocytes. Functional characterization of these two lineages of CD11b(hi) DCs has been hindered by the lack of a widely applicable method to distinguish between them. We performed gene expression analysis of fractionated lung DCs from C57BL/6 mice and found that monocyte-derived DCs (moDCs), including CD11b(hi)Ly-6C(lo) tissue-resident and CD11b(hi)Ly-6C(hi) inflammatory moDCs, express the complement 5a receptor 1/CD88, whereas preDC-derived conventional DCs (cDCs), including CD103(+) and CD11b(hi) cDCs, express dipeptidyl peptidase-4/CD26. Flow cytometric analysis of multiple organs, including the kidney, liver, lung, lymph nodes, small intestine, and spleen, confirmed that reciprocal display of CD88 and CD26 can reliably distinguish FLT3-independent moDCs from FLT3-dependent cDCs in C57BL/6 mice. Similar results were obtained when DCs from BALB/c mice were analyzed. Using this novel approach to study DCs in mediastinal lymph nodes, we observed that most blood-derived lymph node-resident DCs, as well as tissue-derived migratory DCs, are cDCs. Furthermore, cDCs, but not moDCs, stimulated naive T cell proliferation. We anticipate that the use of Abs against CD88 and CD26 to distinguish moDCs and cDCs in multiple organs and mouse strains will facilitate studies aimed at assigning specific functions to distinct DC lineages in immune responses.
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Proliferación Celular/fisiología , Células Dendríticas/inmunología , Dipeptidil Peptidasa 4/inmunología , Regulación de la Expresión Génica/inmunología , Monocitos/inmunología , Receptor de Anafilatoxina C5a/inmunología , Animales , Antígenos CD/genética , Antígenos CD/inmunología , Antígeno CD11b/genética , Antígeno CD11b/inmunología , Células Dendríticas/citología , Dipeptidil Peptidasa 4/genética , Cadenas alfa de Integrinas/genética , Cadenas alfa de Integrinas/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Monocitos/citología , Especificidad de Órganos/genética , Especificidad de Órganos/inmunología , Receptor de Anafilatoxina C5a/genética , Tirosina Quinasa 3 Similar a fms/genética , Tirosina Quinasa 3 Similar a fms/inmunologíaRESUMEN
Mutually exclusive cell fate determination of CD4 helper or CD8 killer T cells occurs in the thymus. These T-cell subsets are not believed to redirect other lineages. Here we showed that retinoic acid and transforming growth factor-ß1 promoted the differentiation of CD8αα T cells from CD4 T cells in a Runx3-dependent manner. These cells were inferred to belong to immunoregulatory populations because subpopulations of CD8αα+TCRαß T cells are known to suppress activated T cells, and mice with Runx3(-/-) T cells showed defects during recovery from experimental allergic encephalomyelitis. Our results demonstrate that CD4 T cells play fundamental roles in controlling immune reactions through promotion and attenuation. We accordingly anticipate that clarifying the mechanisms underlying this process will provide insights leading to autoimmune and immunodeficiency disease therapies.
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Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/metabolismo , Diferenciación Celular , Tejido Linfoide/patología , Receptores de Antígenos de Linfocitos T/metabolismo , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/fisiología , Linfocitos T CD4-Positivos/trasplante , Linfocitos T CD8-positivos/inmunología , Separación Celular , Células Cultivadas , Subunidad alfa 3 del Factor de Unión al Sitio Principal/metabolismo , Subunidad alfa 3 del Factor de Unión al Sitio Principal/fisiología , Citocinas/fisiología , Proteínas de Unión al ADN/genética , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/patología , Citometría de Flujo , Tejido Linfoide/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Bazo/inmunología , Bazo/patología , Linfocitos T Reguladores/metabolismo , TranscriptomaRESUMEN
BACKGROUND: Recent evidence suggests that IL-17 contributes to airway hyperresponsiveness (AHR); however, the mechanisms that suppress the production of this cytokine remain poorly defined. OBJECTIVE: We sought to identify the regulatory cells and molecules that suppress IL-17-dependent allergic airways disease. METHODS: Mice were sensitized by means of airway instillations of ovalbumin together with low levels of LPS. Leukocyte recruitment to the lung and AHR were assessed after daily challenges with aerosolized ovalbumin. Flow cytometry, quantitative PCR, and gene-targeted mice were used to identify naturally arising subsets of regulatory T (Treg) cells and their cytokines required for the suppression of established allergic airway disease. RESULTS: Allergic sensitization through the airway primed both effector and regulatory responses. Effector responses were initially dominant and led to airway inflammation and IL-17-dependent AHR. However, after multiple daily allergen challenges, IL-17 production and AHR decreased, even though pulmonary levels of T(H)17 cells remained high. This loss of AHR was reversible and required the expansion of a Treg cell subset expressing both forkhead box protein 3 and inducible costimulator. These Treg cells also expressed the regulatory cytokines IL-10, TGF-ß, and IL-35. Whereas IL-10 and TGF-ß were dispensable for suppression of AHR, IL-35 was required. CONCLUSION: IL-35 production by inducible costimulator-positive Treg cells can suppress IL-17 production and thereby reverse established, IL-17-dependent AHR in mice. Targeting this pathway might therefore be of therapeutic value for treating allergic asthma in human subjects.
Asunto(s)
Asma/inmunología , Proteína Coestimuladora de Linfocitos T Inducibles/metabolismo , Interleucina-17/biosíntesis , Interleucinas/biosíntesis , Linfocitos T Reguladores/inmunología , Alérgenos/inmunología , Animales , Asma/metabolismo , Recuento de Linfocito CD4 , Citocinas/biosíntesis , Modelos Animales de Enfermedad , Factores de Transcripción Forkhead/metabolismo , Tolerancia Inmunológica , Ligando Coestimulador de Linfocitos T Inducibles/metabolismo , Interleucina-17/farmacología , Pulmón/inmunología , Pulmón/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Unión Proteica , Linfocitos T Reguladores/metabolismo , Células Th17/inmunologíaRESUMEN
PURPOSE: The purpose of this study was to evaluate the frequency of tracheobronchomalacia (TBM) associated with pulmonary emphysema with paired inspiratory-expiratory multidetector computed tomography (MDCT) using a low-dose technique. MATERIALS AND METHODS: This study included 56 consecutive patients (55 men, 1 woman; mean age 68.9 years) with pulmonary emphysema who had undergone paired inspiratory-expiratory CT scanning with a low-dose technique (40 mA). All images were retrospectively examined by two thoracic radiologists in a blinded fashion. The diagnosis of TBM was based on the standard criterion of >50% reduction in the cross-sectional area of the tracheobronchial lumen at the end-expiratory phase. A mild TBM criterion of >30% reduction was also reviewed. All patients underwent pulmonary function tests. The relation between the forced expiratory volume in 1 s (FEV(1.0%)) and TBM was statistically analyzed. RESULTS: Four (7.1%) and eight (14.3%) patients were diagnosed as TBM based on the standard and mild criteria, respectively. In four patients, the percentages of luminal narrowing were 63.4% and 51.2%, respectively for tracheomalacia and 59.2% and 62.0%, respectively, for bronchomalacia. The FEV(1.0%) values between patients with and without TBM showed no statistical difference. CONCLUSION: The incidence of TBM associated with pulmonary emphysema was 7.1% with the standard criterion. It is possible that TBM has been underdiagnosed in a number of patients with pulmonary emphysema.
Asunto(s)
Enfisema Pulmonar/complicaciones , Traqueobroncomalacia/epidemiología , Traqueobroncomalacia/etiología , Anciano , Femenino , Humanos , Incidencia , Japón/epidemiología , Masculino , Enfisema Pulmonar/diagnóstico por imagen , Dosis de Radiación , Pruebas de Función Respiratoria , Estudios Retrospectivos , Estadísticas no Paramétricas , Tomografía Computarizada por Rayos X , Traqueobroncomalacia/diagnóstico por imagenRESUMEN
Sorghum, Sorghum bicolor (L.) Moench, is the fifth most important cereal crop in the world. In this study, we identified the IgE production-suppressing activity of white sorghum bran extracts. White sorghum is one of the genotypes of sorghum. White sorghum bran extracts in 10 mM sodium phosphate buffer (pH 7.4) suppressed IgE production in human myeloma cell line U266. The extracts suppressed IgE production by decreasing mRNA transcription level of IgE, but they did not affect IgA or IgG production of mice splenocytes in vitro. Heat treatment and trypsin digestion did not affect IgE production-suppressing activity. The white sorghum bran extracts were fractionated by ultrafiltration, and the molecular weight of the active substance was estimated to be less than 1,000.