RESUMO
Understanding and controlling nanoscale interface phenomena, such as band bending and secondary phase formation, is crucial for electronic device optimization. In granular metal (GM) studies, where metal nanoparticles are embedded in an insulating matrix, the importance of interface phenomena is frequently neglected. We demonstrate that GMs can serve as an exemplar system for evaluating the role of secondary phases at interfaces through a combination of x-ray photoemission spectroscopy (XPS) and electrical transport studies. We investigated SiNxas an alternative to more commonly used oxide-insulators, as SiNx-based GMs may enable high temperature applications when paired with refractory metals. Comparing Co-SiNxand Mo-SiNxGMs, we found that, in the tunneling-dominated insulating regime, Mo-SiNxhad reduced metal-silicide formation and orders-of-magnitude lower conductivity. XPS measurements indicate that metal-silicide and metal-nitride formation are mitigatable concerns in Mo-SiNx. Given the metal-oxide formation seen in other GMs, SiNxis an appealing alternative for metals that readily oxidize. Furthermore, SiNxprovides a path to metal-nitride nanostructures, potentially useful for various applications in plasmonics, optics, and sensing.
RESUMO
Eosinophil activities are often linked with allergic diseases such as asthma and the pathologies accompanying helminth infection. These activities have been hypothesized to be mediated, in part, by the release of cationic proteins stored in the secondary granules of these granulocytes. The majority of the proteins stored in these secondary granules (by mass) are major basic protein 1 (MBP-1) and eosinophil peroxidase (EPX). Unpredictably, a knockout approach targeting the genes encoding these proteins demonstrated that, unlike in mice containing a single deficiency of only MBP-1 or EPX, the absence of both granule proteins resulted in the near complete loss of peripheral blood eosinophils with no apparent impact on any other hematopoietic lineage. Moreover, the absence of MBP-1 and EPX promoted a concomitant loss of eosinophil lineage-committed progenitors in the marrow, identifying a specific blockade in eosinophilopoiesis as the causative event. Significantly, this blockade of eosinophilopoiesis is also observed in ex vivo cultures of marrow progenitors and is not rescued in vivo by adoptive bone marrow engraftment, suggesting a cell-autonomous defect in marrow progenitors. These observations implicate a role for granule protein gene expression as a regulator of eosinophilopoiesis and provide another strain of mice congenitally deficient of eosinophils.
Assuntos
Proteína Básica Maior de Eosinófilos/fisiologia , Peroxidase de Eosinófilo/fisiologia , Eosinófilos/fisiologia , Mielopoese/genética , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Células da Medula Óssea/fisiologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Proteína Básica Maior de Eosinófilos/genética , Proteína Básica Maior de Eosinófilos/metabolismo , Peroxidase de Eosinófilo/genética , Peroxidase de Eosinófilo/metabolismo , Eosinófilos/efeitos dos fármacos , Eosinófilos/metabolismo , Interleucina-5/farmacologia , Contagem de Leucócitos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mielopoese/efeitos dos fármacos , Mielopoese/fisiologiaRESUMO
The respective life histories of human subjects and mice are well defined and describe a unique story of evolutionary conservation extending from sequence identity within the genome to the underpinnings of biochemical, cellular, and physiologic pathways. As a consequence, the hematopoietic lineages of both species are invariantly maintained, each with identifiable eosinophils. This canonical presence nonetheless does not preclude disparities between human and mouse eosinophils, their effector functions, or both. Indeed, many books and reviews dogmatically highlight differences, providing a rationale to discount the use of mouse models of human eosinophilic diseases. We suggest that this perspective is parochial and ignores the wealth of available studies and the consensus of the literature that overwhelming similarities (and not differences) exist between human and mouse eosinophils. The goal of this review is to summarize this literature and in some cases provide experimental details comparing and contrasting eosinophils and eosinophil effector functions in human subjects versus mice. In particular, our review will provide a summation and an easy-to-use reference guide to important studies demonstrating that although differences exist, more often than not, their consequences are unknown and do not necessarily reflect inherent disparities in eosinophil function but instead species-specific variations. The conclusion from this overview is that despite nominal differences, the vast similarities between human and mouse eosinophils provide important insights as to their roles in health and disease and, in turn, demonstrate the unique utility of mouse-based studies with an expectation of valid extrapolation to the understanding and treatment of patients.
Assuntos
Eosinófilos/fisiologia , Animais , Degranulação Celular , Proteína Catiônica de Eosinófilo/fisiologia , Peroxidase de Eosinófilo/fisiologia , Evolução Molecular , Glicoproteínas/fisiologia , Hematopoese , Humanos , Lisofosfolipase/fisiologia , CamundongosRESUMO
The genetic and physiological similarities between mice and humans have focused considerable attention on rodents as potential models of human health and disease. Together with the wealth of resources, knowledge, and technologies surrounding the mouse as a model system, these similarities have propelled this species to the forefront of biomedical research. The advent of genomic manipulation has quickly led to the creation and use of genetically engineered mice as powerful tools for cutting edge studies of human disease research including the discovery, refinement, and utility of many currently available therapeutic regimes. In particular, the creation of genetically modified mice as models of human disease has remarkably changed our ability to understand the molecular mechanisms and cellular pathways underlying disease states. Moreover, the mouse models resulting from gene transfer technologies have been important components correlating an individual's gene expression profile to the development of disease pathologies. The objective of this review is to provide physician-scientists with an expansive historical and logistical overview of the creation of mouse models of human disease through gene transfer technologies. Our expectation is that this will facilitate on-going disease research studies and may initiate new areas of translational research leading to enhanced patient care.
Assuntos
Modelos Animais de Doenças , Regulação da Expressão Gênica , Técnicas de Introdução de Genes/métodos , Técnicas de Inativação de Genes/métodos , Engenharia Genética/métodos , Animais , Cromossomos de Mamíferos/genética , Cromossomos de Mamíferos/metabolismo , Marcação de Genes/métodos , Técnicas de Transferência de Genes , Genoma , Humanos , Camundongos , Camundongos Knockout , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Homologia de Sequência do Ácido Nucleico , Especificidade da EspécieRESUMO
BACKGROUND & AIMS: Eosinophilic esophagitis (EoE) is characterized by medically/surgically-resistant gastroesophageal reflux symptoms and dense squamous eosinophilia. Studies suggest that histologic assessment of esophageal eosinophilia alone cannot reliably separate patients with EoE from those with gastroesophageal reflux disease (GERD). Our goal was to develop an assay to identify EoE patients and perhaps differentiate EoE from other causes of esophageal eosinophilia. METHODS: A monoclonal antibody specific for an eosinophil secondary granule protein (eosinophil peroxidase [EPX]) was developed and shown to specifically identify intact eosinophils and detect eosinophil degranulation in formalin-fixed specimens. A histopathologic scoring algorithm was developed to analyze data from patient evaluations; the utility of this algorithm was assessed by using archived esophageal tissues from patients with known diagnoses of EoE and GERD as well as controls from 2 tertiary care centers. RESULTS: Intraobserver/interobserver blinded evaluations demonstrated a significant difference (P < .001) between scores of samples taken from control subjects, from patients with esophageal eosinophilia who had a diagnosis of EoE, and from patients with GERD (P < .001). This algorithm also was able to identify patients whose clinical course was suggestive of a diagnosis of EoE, but that nonetheless failed to reach the critical threshold number of > or =15 eosinophils in a high-power (40x) microscopy field. CONCLUSIONS: A novel immunohistochemical scoring system was developed to address an unmet medical need to differentiate histologic specimens from patients with EoE relative to those with GERD. The availability of a unique anti-EPX-specific monoclonal antibody, combined with the ease/rapidity of this staining method and scoring system, will provide a valuable strategy for the assessment of esophageal eosinophilia.
Assuntos
Biópsia , Eosinofilia/diagnóstico , Eosinofilia/patologia , Esofagite/diagnóstico , Esofagite/patologia , Índice de Gravidade de Doença , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Animais , Criança , Pré-Escolar , Diagnóstico Diferencial , Eosinofilia/imunologia , Esofagite/imunologia , Refluxo Gastroesofágico/diagnóstico , Refluxo Gastroesofágico/patologia , Humanos , Imuno-Histoquímica/métodos , Lactente , Camundongos , Pessoa de Meia-Idade , Coloração e RotulagemRESUMO
Allergen-induced respiratory inflammation facilitates and/or elicits the extravasation of proinflammatory leukocytes by well-understood mechanisms that mediate the movement of multiple cell types. The nonspecific character of these pathways led us to hypothesize that circulating cancer cells use similar mechanisms, promoting secondary tumor formation at distal sites. To test this hypothesis, the frequency of metastasis to the lung as a function of allergic pulmonary inflammation was assessed following the i.v. injection of B16-F10 melanoma cells in mice. These studies showed that allergen-induced pulmonary inflammation resulted in a >3-fold increase in lung metastases. This increase was dependent on CD4(+) T-cell activities; however, it occurred independent of the induced eosinophilia associated with allergen provocation. Interventional strategies showed that existing therapeutic modalities for asthma, such as inhaled corticosteroids, were sufficient to block the enhanced pulmonary recruitment of cancer cells from circulation. Additional mechanistic studies further suggested that the ability of circulating cancer cells to extravasate to surrounding lung tissues was linked to the activation of the vascular endothelium via one or more Galpha(i)-coupled receptors. Interestingly, a survey of a clinical breast cancer surgical database showed that the incidence of asthma was higher among patients with lung metastases. Thus, our data show that allergic respiratory inflammation may represent a risk factor for the development of lung metastases and suggest that amelioration of the pulmonary inflammation associated with asthma will have a direct and immediate benefit to the 7% to 8% of breast cancer patients with this lung disease.
Assuntos
Asma/complicações , Neoplasias Pulmonares/secundário , Células Neoplásicas Circulantes , Animais , Neoplasias da Mama/complicações , Neoplasias da Mama/patologia , Budesonida/farmacologia , Linfócitos T CD4-Positivos/imunologia , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Neoplasias do Colo/patologia , Células Endoteliais/fisiologia , Eosinofilia/complicações , Feminino , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/fisiologia , Humanos , Pulmão/patologia , Melanoma Experimental/secundário , Camundongos , Camundongos Endogâmicos C57BL , Molécula 1 de Adesão de Célula Vascular/fisiologiaRESUMO
Mouse models of allergen provocation and/or transgenic gene expression have provided significant insights regarding the cellular, molecular, and immune responses linked to the pathologies occurring as a result of allergic respiratory inflammation. Nonetheless, the inability to replicate the eosinophil activities occurring in patients with asthma has limited their usefulness to understand the larger role(s) of eosinophils in disease pathologies. These limitations have led us to develop an allergen-naive double transgenic mouse model that expresses IL-5 systemically from mature T cells and eotaxin-2 locally from lung epithelial cells. We show that these mice develop several pulmonary pathologies representative of severe asthma, including structural remodeling events such as epithelial desquamation and mucus hypersecretion leading to airway obstruction, subepithelial fibrosis, airway smooth muscle hyperplasia, and pathophysiological changes exemplified by exacerbated methacholine-induced airway hyperresponsiveness. More importantly, and similar to human patients, the pulmonary pathologies observed are accompanied by extensive eosinophil degranulation. Genetic ablation of all eosinophils from this double transgenic model abolished the induced pulmonary pathologies, demonstrating that these pathologies are a consequence of one or more eosinophil effector functions.
Assuntos
Asma/imunologia , Quimiocinas CC/metabolismo , Eosinófilos/imunologia , Interleucina-5/metabolismo , Eosinofilia Pulmonar/imunologia , Animais , Asma/genética , Asma/patologia , Líquido da Lavagem Broncoalveolar/imunologia , Movimento Celular , Quimiocina CCL24 , Quimiocinas CC/genética , Modelos Animais de Doenças , Peroxidase de Eosinófilo/análise , Eosinófilos/diagnóstico por imagem , Eosinófilos/enzimologia , Humanos , Interleucina-5/genética , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Transgênicos , Pneumonia/genética , Pneumonia/imunologia , Eosinofilia Pulmonar/genética , Eosinofilia Pulmonar/patologia , UltrassonografiaRESUMO
The trafficking of leukocytes from the blood to sites of inflammation is the cumulative result of receptor-ligand-mediated signaling events associated with the leukocytes themselves as well as with the underlying vascular endothelium. Our data show that Galpha(i) signaling pathways in the vascular endothelium regulate a critical step required for leukocyte diapedesis. In vivo studies using knockout mice demonstrated that a signaling event in a non-lymphohematopoietic compartment of the lung prevented the recruitment of proinflammatory leukocytes. Intravital microscopy showed that blockade was at the capillary endothelial surface and ex vivo studies of leukocyte trafficking demonstrated that a Galpha(i)-signaling event in endothelial cells was required for transmigration. Collectively, these data suggest that specific Galpha(i2)-mediated signaling between endothelial cells and leukocytes is required for the extravasation of leukocytes and for tissue-specific accumulation.
Assuntos
Endotélio Vascular/metabolismo , Subunidade alfa Gi2 de Proteína de Ligação ao GTP/metabolismo , Leucócitos/metabolismo , Transdução de Sinais , Alérgenos/metabolismo , Animais , Endotélio Vascular/citologia , Endotoxinas/metabolismo , Eosinófilos/metabolismo , Inflamação , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neutrófilos/metabolismo , Toxina Pertussis/farmacologiaRESUMO
Insects used in research have traditionally been housed and cared for in the investigator's laboratory. Centralized colony maintenance may be advantageous, but presents unique challenges to animal care staff members, who are more familiar with vertebrate research animals. To fill this potential knowledge gap, the authors share the procedures they have developed at Arizona State University for the housing, husbandry, and breeding of grasshoppers used in research and teaching.
Assuntos
Criação de Animais Domésticos/métodos , Gafanhotos/fisiologia , Animais , Animais de Laboratório , Gafanhotos/crescimento & desenvolvimentoRESUMO
The identification and characterization of mouse basophils have historically been hampered by the extreme rarity of this cell type. Virtually no photomicrographs of hematologically stained (eg, Wright-Giemsa) examples of mouse basophils exist in the literature. However, 4 recent studies in the past 2 years have used flow cytometry and a defined set of cell-surface markers to identify and subsequently isolate mouse "basophils," including the publication of stained cytospin preparations of these cells. Surprisingly, a reevaluation of the data from all 4 of the studies revealed several issues of concern that suggest that the cells under study are not necessarily basophils. Nonetheless, we propose that these studies do provide the foundation for a reevaluation of the defining characteristics of a basophil and/or provide support for the provocative conclusion that a new previously overlooked leukocyte subtype has been identified. The purpose of this commentary is to revisit these previously published studies, highlight the relevant issues, and provide a different perspective in the hope of developing a consensus within the research community as to the true identity of the "basophils" described in these studies.
Assuntos
Basófilos/citologia , Camundongos , Animais , Antígenos de Superfície , Biomarcadores , Citometria de Fluxo , Leucócitos/citologiaRESUMO
Tumor-associated eosinophilia has been observed in numerous human cancers and several tumor models in animals; however, the details surrounding this eosinophilia remain largely undefined and anecdotal. We used a B16-F10 melanoma cell injection model to demonstrate that eosinophil infiltration of tumors occurred from the earliest palpable stages with significant accumulations only in the necrotic and capsule regions. Furthermore, the presence of diffuse extracellular matrix staining for eosinophil major basic protein was restricted to the necrotic areas of tumors, indicating that eosinophil degranulation was limited to this region. Antibody-mediated depletion of CD4+ T cells and adoptive transfer of eosinophils suggested, respectively, that the accumulation of eosinophils is not associated with T helper cell type 2-dependent immune responses and that recruitment is a dynamic, ongoing process, occurring throughout tumor growth. Ex vivo migration studies have identified what appears to be a novel chemotactic factor(s) released by stressed/dying melanoma cells, suggesting that the accumulation of eosinophils in tumors occurs, in part, through a unique mechanism dependent on a signal(s) released from areas of necrosis. Collectively, these studies demonstrate that the infiltration of tumors by eosinophils is an early and persistent response that is spatial-restricted. It is more important that these data also show that the mechanism(s) that elicit this host response occur, independent of immune surveillance, suggesting that eosinophils are part of an early inflammatory reaction at the site of tumorigenesis.
Assuntos
Quimiotaxia/fisiologia , Eosinófilos/imunologia , Inflamação/imunologia , Melanoma Experimental/imunologia , Animais , Antígenos CD4/imunologia , Linfócitos T CD4-Positivos/imunologia , Células Cultivadas , Fatores Quimiotáticos/metabolismo , Quimiotaxia/efeitos dos fármacos , Meios de Cultivo Condicionados/química , Meios de Cultivo Condicionados/farmacologia , Eosinofilia/etiologia , Eosinofilia/fisiopatologia , Eosinófilos/transplante , Vigilância Imunológica , Imunoterapia Adotiva , Inflamação/patologia , Injeções Subcutâneas , Interleucina-5/genética , Depleção Linfocítica , Melanoma Experimental/complicações , Melanoma Experimental/metabolismo , Melanoma Experimental/patologia , Melanoma Experimental/terapia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Confocal , Necrose , Transplante de Neoplasias , Células Th2/imunologiaRESUMO
We have observed decreased size and increased mortality rates in interleukin 5 (IL-5)-deficient mice versus IL-5-heterozygous and wild-type mice and have sought to define these differences. IL-5-deficient mice nursed by IL-5 deficient mothers were notably underweight, with a high percentage of preweaning mortality. In contrast, IL-5-deficient mice nursed by IL-5-sufficient foster mothers from birth were well-developed and robust at weaning, with a relatively low percentage of preweaning mortality. Mammary tissues from IL-5-deficient females at various landmark stages throughout life were prepared for microscopic assessment. When compared with mammary tissue from normal mice, that from IL-5-deficient dams appeared to have fewer terminal end buds, less well-developed branching of the mammary ducts, and lower overall density of mammary gland structures. The molecular and cellular bases for the differences in mammary gland development in IL-5-deficient mice relative to wild-type animals remains unknown. Under consideration are the roles that IL-5 and eosinophil granulocytes (the primary cell responsive to IL-5) may have in mammary gland development.
Assuntos
Hospedeiro Imunocomprometido , Interleucina-5/deficiência , Lactação/imunologia , Longevidade/imunologia , Animais , Peso Corporal/genética , Peso Corporal/imunologia , Deleção de Genes , Hospedeiro Imunocomprometido/genética , Hospedeiro Imunocomprometido/imunologia , Interleucina-5/genética , Interleucina-5/imunologia , Lactação/genética , Longevidade/genética , Glândulas Mamárias Animais/patologia , Camundongos , Camundongos Knockout , DesmameRESUMO
Eosinophils are often dominant inflammatory cells present in the lungs of asthma patients. Nonetheless, the role of these leukocytes remains poorly understood. We have created a transgenic line of mice (PHIL) that are specifically devoid of eosinophils, but otherwise have a full complement of hematopoietically derived cells. Allergen challenge of PHIL mice demonstrated that eosinophils were required for pulmonary mucus accumulation and the airway hyperresponsiveness associated with asthma. The development of an eosinophil-less mouse now permits an unambiguous assessment of a number of human diseases that have been linked to this granulocyte, including allergic diseases, parasite infections, and tumorigenesis.
Assuntos
Asma/patologia , Asma/fisiopatologia , Eosinófilos/fisiologia , Pulmão/patologia , Pulmão/fisiopatologia , Alérgenos/imunologia , Animais , Asma/imunologia , Toxina Diftérica/genética , Peroxidase de Eosinófilo , Marcação de Genes , Contagem de Leucócitos , Pulmão/imunologia , Camundongos , Camundongos Transgênicos , Modelos Animais , Muco/metabolismo , Ovalbumina/imunologia , Fragmentos de Peptídeos/genética , Peroxidases/genética , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/patologiaRESUMO
Asthma and mouse models of allergic respiratory inflammation are invariably associated with a pulmonary eosinophilia; however, this association has remained correlative. In this report, a causative relationship between eosinophils and allergen-provoked pathologies was established using eosinophil adoptive transfer. Eosinophils were transferred directly into the lungs of either naive or OVA-treated IL-5(-/-) mice. This strategy resulted in a pulmonary eosinophilia equivalent to that observed in OVA-treated wild-type animals. A concomitant consequence of this eosinophil transfer was an increase in Th2 bronchoalveolar lavage cytokine levels and the restoration of intracellular epithelial mucus in OVA-treated IL-5(-/-) mice equivalent to OVA-treated wild-type levels. Moreover, the transfer also resulted in the development of airway hyperresponsiveness. These pulmonary changes did not occur when eosinophils were transferred into naive IL-5(-/-) mice, eliminating nonspecific consequences of the eosinophil transfer as a possible explanation. Significantly, administration of OVA-treated IL-5(-/-) mice with GK1.5 (anti-CD4) Abs abolished the increases in mucus accumulation and airway hyperresponsiveness following adoptive transfer of eosinophils. Thus, CD4(+) T cell-mediated inflammatory signals as well as signals derived from eosinophils are each necessary, yet alone insufficient, for the development of allergic pulmonary pathology. These data support an expanded view of T cell and eosinophil activities and suggest that eosinophil effector functions impinge directly on lung function.
Assuntos
Alérgenos/imunologia , Eosinófilos/imunologia , Eosinófilos/patologia , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/patologia , Transferência Adotiva , Aerossóis , Alérgenos/administração & dosagem , Animais , Hiper-Reatividade Brônquica/genética , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/patologia , Linfócitos T CD4-Positivos/imunologia , Modelos Animais de Doenças , Eosinófilos/transplante , Interleucina-5/deficiência , Interleucina-5/genética , Intubação Intratraqueal , Pulmão/imunologia , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Muco/metabolismo , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Eosinofilia Pulmonar/genética , Eosinofilia Pulmonar/imunologia , Eosinofilia Pulmonar/patologia , Hipersensibilidade Respiratória/genética , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologiaRESUMO
The animal research literature devotes much attention to the function of clothing and personal protective equipment in preventing transmission of zoonotic diseases from animal to human, but has said comparatively little about the potential for transmission of disease from human to animal. The authors consider appropriate garbing standards for personnel charged with various tasks within the animal facility, emphasizing the protection of animals from potential health threats.
Assuntos
Doença Iatrogênica/prevenção & controle , Ciência dos Animais de Laboratório/normas , Pessoal de Laboratório Médico , Roupa de Proteção , Animais , Feminino , Humanos , Ciência dos Animais de Laboratório/métodos , Masculino , ZoonosesRESUMO
A strategy to deplete eosinophils from the lungs of ovalbumin (OVA)-sensitized/challenged mice was developed using antibody-mediated depletion. Concurrent administration [viz. the peritoneal cavity (systemic) and as an aerosol to the lung (local)] of a rat anti-mouse CCR3 monoclonal antibody resulted in the abolition of eosinophils from the lung such that the airway lumen was essentially devoid of eosinophils. Moreover, perivascular/peribronchial eosinophil numbers were reduced to levels indistinguishable from saline-challenged animals. This antibody-mediated depletion was not accompanied by effects on any other leukocyte population, including, but not limited to, T cells and mast cells/basophils. In addition, no effects were observed on other underlying allergic inflammatory responses in OVA-treated mice, including OVA-specific immunoglobulin production as well as T cell-dependent elaboration of Th2 cytokines. The ablation of virtually all pulmonary eosinophils in OVA-treated mice (i.e., without concurrent effects on T cell activities) resulted in a significant decrease in mucus accumulation and abolished allergen-induced airway hyperresponsiveness. These data demonstrate a direct causative relationship between allergen-mediated pulmonary pathologies and eosinophils.
Assuntos
Alérgenos/imunologia , Eosinófilos/fisiologia , Pulmão/imunologia , Pulmão/fisiopatologia , Ovalbumina/imunologia , Animais , Anticorpos Monoclonais/farmacologia , Formação de Anticorpos , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/fisiopatologia , Contagem de Células , Citocinas/metabolismo , Eosinófilos/efeitos dos fármacos , Células Caliciformes/patologia , Imunoglobulinas/biossíntese , Pulmão/patologia , Linfócitos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Muco/metabolismo , Ratos , Receptores CCR3 , Receptores de Quimiocinas/imunologia , Células Th2/metabolismoRESUMO
The ashen (ash) mouse, a model for Hermansky-Pudlak syndrome (HPS) and for a subset of patients with Griscelli syndrome, presents with hypopigmentation, prolonged bleeding times, and platelet storage pool deficiency due to a mutation which abrogates expression of the Rab27a protein. Platelets of mice with the ashen mutation on the C3H/HeSnJ inbred strain background have greatly reduced amounts of dense granule components such as serotonin and adenine nucleotides though near-normal numbers of dense granules as enumerated by the dense granule-specific fluorescent dye mepacrine. Thus, essentially normal numbers of platelet dense granules are produced but the granule interiors are abnormal. Collagen-mediated aggregation of mutant platelets is significantly depressed. No abnormalities in the concentrations or secretory rates of 2 other major platelet granules, lysosomes and alpha granules, were apparent. Similarly, no platelet ultrastructural alterations other than those involving dense granules were detected. Therefore, Rab27a regulates the synthesis and secretion of only one major platelet organelle, the dense granule. There were likewise no mutant effects on levels or secretion of lysosomal enzymes of several other tissues. Together with other recent analyses of the ashen mouse, these results suggest a close relationship between platelet dense granules, melanosomes of melanocytes and secretory lysosomes of cytotoxic T lymphocytes, all mediated by Rab27a. Surprisingly, the effects of the ashen mutation on platelet-dense granule components, platelet aggregation, and bleeding times were highly dependent on genetic background. This suggests that bleeding tendencies may likewise vary among patients with Griscelli syndrome and HPS with Rab27a mutations.
Assuntos
Plaquetas/ultraestrutura , Grânulos Citoplasmáticos/efeitos dos fármacos , Síndrome de Hermanski-Pudlak/genética , Proteínas rab de Ligação ao GTP/fisiologia , Difosfato de Adenosina/deficiência , Trifosfato de Adenosina/deficiência , Animais , Plaquetas/química , Grânulos Citoplasmáticos/química , Modelos Animais de Doenças , Predisposição Genética para Doença , Hemorragia/etiologia , Hemorragia/genética , Síndrome de Hermanski-Pudlak/complicações , Camundongos , Camundongos Mutantes , Deficiência do Pool Plaquetário/genética , Serotonina/deficiência , Proteínas rab de Ligação ao GTP/genética , Proteínas rab de Ligação ao GTP/farmacologia , Proteínas rab27 de Ligação ao GTPRESUMO
Several single gene pigment mutants of inbred C57BL/6J mice display a triad of subcellular granule-associated defects: oculocutaneous pigment dilution, prolonged bleeding due to defects in platelet dense granules, and abnormal lysosomes. These features also characterize Hermansky-Pudlak Syndrome (HPS), making these mice relevant animal models for HPS. Mice of one mutant strain, pallid, in addition to the hallmark triad of signs, also exhibit age-dependent lung pathology. Respiratory system mechanics showed that the age-dependent histopathology of pallid mice was accompanied by a decrease in lung reactance. Furthermore, the possibility that pallid pulmonary pathology may result from persistent inflammation due to microhemorrhage owing to the platelet defect was examined. Hematopoietic reconstitution of pallid mice with marrow from normal C57BL/6J donors did not prevent the development of the pulmonary histopathology or respiratory system mechanics characteristic of the pallid genotype. Similarly, wild-type mice 12 months after engraftment with pallid marrow did not develop pallid-like pulmonary histopathology or respiratory system mechanics. Thus, pallid-associated pulmonary functional and structural pathologies are not linked to the marrow (bleeding) genotype, but instead are the result of an age-dependent process resulting from a defect(s) in one or more nonhematopoietic cell types.
Assuntos
Síndrome de Hermanski-Pudlak/genética , Pulmão/patologia , Transtornos da Pigmentação/genética , Transtornos da Pigmentação/patologia , Animais , Modelos Animais de Doenças , Feminino , Hematopoese , Transplante de Células-Tronco Hematopoéticas , Síndrome de Hermanski-Pudlak/patologia , Síndrome de Hermanski-Pudlak/fisiopatologia , Humanos , Pulmão/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Mutação , Transtornos da Pigmentação/fisiopatologia , Mecânica RespiratóriaRESUMO
The complexity and magnitude of interactions leading to the selective infiltration of eosinophils in response to inhaled allergens are formidable obstacles to a larger understanding of the pulmonary pathology associated with allergic asthma. This study uses knockout mice to demonstrate a novel function for the heterotrimeric G protein, G(q), in the regulation of pulmonary eosinophil recruitment. In the absence of G(q) signaling, eosinophils failed to accumulate in the lungs following allergen challenge. These studies demonstrate that the inhibition of eosinophil accumulation in the airways is attributed to the failure of hemopoietically derived cells to elaborate GM-CSF in the airways. The data suggest that activation of a G(q)-coupled receptor(s) on resident leukocytes in the lung elicits expression of GM-CSF, which, in turn, is required for allergen-induced pulmonary eosinophilia, identifying a novel pathway of eosinophil-associated effector functions leading to pulmonary pathology in diseases such as asthma.