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1.
Allergy ; 71(4): 567-70, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26645423

RESUMEN

The objective of the study was to compare nasal, pharyngeal, and sputum eosinophil peroxidase (EPX) levels with induced sputum eosinophil percentage in 10 adults with poorly controlled asthma and 10 normal controls. EPX was measured using an ELISA and normalized for grams of protein for nasal and pharynx specimens and for mL-gram of protein for sputum. Sputum EPX levels were statistically different between asthma and control subjects (P = 0.024). EPX levels measured in the nasal and pharyngeal swab samples derived from the same patients were also different between asthma and control subjects, each displaying a high degree of significance (P = 0.002). Spearman's correlation coefficients for nasal EPX and pharyngeal EPX levels compared to induced sputum eosinophil percentage were 0.81 (P = 0.0007) and 0.78 (P = 0.0017), respectively. Thus, there is a strong association in a given patient between both nasal and pharyngeal EPX levels and the eosinophil percentage of induced sputum.


Asunto(s)
Asma/diagnóstico , Asma/metabolismo , Peroxidasa del Eosinófilo/metabolismo , Eosinófilos/enzimología , Eosinófilos/patología , Mucosa Nasal/metabolismo , Faringe/metabolismo , Esputo/enzimología , Adulto , Asma/tratamiento farmacológico , Estudios de Casos y Controles , Manejo de la Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana Edad
2.
Allergy ; 70(9): 1148-59, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26009788

RESUMEN

BACKGROUND: Eosinophils are hallmark cells of allergic Th2 respiratory inflammation. However, the relative importance of eosinophil activation and the induction of effector functions such as the expression of IL-13 to allergic Th2 pulmonary disease remain to be defined. METHODS: Wild-type or cytokine-deficient (IL-13(-/-) or IL-4(-/-) ) eosinophils treated with cytokines (GM-CSF, IL-4, IL-33) were adoptively transferred into eosinophil-deficient recipient mice subjected to allergen provocation using established models of respiratory inflammation. Allergen-induced pulmonary changes were assessed. RESULTS: In contrast to the transfer of untreated blood eosinophils to the lungs of recipient eosinophil deficient mice, which induced no immune/inflammatory changes either in the lung or in the lung draining lymph nodes (LDLN), pretreatment of blood eosinophils with GM-CSF prior to transfer elicited trafficking of these eosinophils to LDLN. In turn, these LDLN eosinophils elicited the accumulation of dendritic cells and CD4(+) T cells to these same LDLNs without inducing pulmonary inflammation. However, exposure of eosinophils to GM-CSF, IL-4, and IL-33 prior to transfer induced not only immune events in the LDLN, but also allergen-mediated increases in airway Th2 cytokine/chemokine levels, the subsequent accumulation of CD4(+) T cells as well as alternatively activated (M2) macrophages, and the induction of pulmonary histopathologies. Significantly, this allergic respiratory inflammation was dependent on eosinophil-derived IL-13, whereas IL-4 expression by eosinophils had no significant role. CONCLUSION: The data demonstrate the differential activation of eosinophils as a function of cytokine exposure and suggest that eosinophil-specific IL-13 expression by activated cells is a necessary component of the subsequent allergic Th2 pulmonary pathologies.


Asunto(s)
Eosinófilos/inmunología , Eosinófilos/metabolismo , Hipersensibilidad/inmunología , Hipersensibilidad/metabolismo , Interleucina-13/metabolismo , Células Th2/inmunología , Células Th2/metabolismo , Alérgenos/inmunología , Animales , Células Cultivadas , Citocinas/metabolismo , Citocinas/farmacología , Modelos Animales de Enfermedad , Eosinófilos/efectos de los fármacos , Femenino , Hipersensibilidad/genética , Hipersensibilidad/patología , Interleucina-13/genética , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Transgénicos , Ovalbúmina/inmunología , Fenotipo
3.
Clin Exp Allergy ; 44(9): 1119-36, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24961290

RESUMEN

The role of eosinophils in the progression and resolution of allergic respiratory inflammation is poorly defined despite the commonality of their presence and in some cases their use as a biomarker for disease severity and/or symptom control. However, this ambiguity belies the wealth of insights that have recently been gained through the use of eosinophil-deficient/attenuated strains of mice that have demonstrated novel immunoregulatory and remodelling/repair functions for these cells in the lung following allergen provocation. Specifically, studies of eosinophil-deficient mice suggest that eosinophils contribute to events occurring in the lungs following allergen provocation at several key moments: (i) the initiating phase of events leading to Th2-polarized pulmonary inflammation, (ii) the suppression Th1/Th17 pathways in lung-draining lymph nodes, (iii) the recruitment of effector Th2 T cells to the lung, and finally, (iv) mechanisms of inflammatory resolution that re-establish pulmonary homoeostasis. These suggested functions have recently been confirmed and expanded upon using allergen provocation of an inducible eosinophil-deficient strain of mice (iPHIL) that demonstrated an eosinophil-dependent mechanism(s) leading to Th2 dominated immune responses in the presence of eosinophils in contrast to neutrophilic as well as mixed Th1/Th17/Th2 variant phenotypes in the absence of eosinophils. These findings highlighted that eosinophils are not exclusively downstream mediators controlled by T cells, dendritic cells (DC) and/or innate lymphocytic cells (ILC2). Instead, eosinophils appear to be more aptly described as significant contributors in complex interrelated pathways that lead to pulmonary inflammation and subsequently promote resolution and the re-establishment of homoeostatic baseline. In this review, we summarize and put into the context the evolving hypotheses that are now expanding our understanding of the roles eosinophils likely have in the lung following allergen provocation.


Asunto(s)
Eosinófilos/inmunología , Hipersensibilidad Respiratoria/inmunología , Traslado Adoptivo , Alérgenos/inmunología , Animales , Asma/diagnóstico , Asma/inmunología , Asma/metabolismo , Modelos Animales de Enfermedad , Eosinófilos/metabolismo , Humanos , Ratones , Fenotipo , Hipersensibilidad Respiratoria/diagnóstico , Hipersensibilidad Respiratoria/metabolismo , Hipersensibilidad Respiratoria/terapia , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo
4.
Allergy ; 69(3): 315-27, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24266710

RESUMEN

BACKGROUND: The importance and specific role(s) of eosinophils in modulating the immune/inflammatory phenotype of allergic pulmonary disease remain to be defined. Established animal models assessing the role(s) of eosinophils as contributors and/or causative agents of disease have relied on congenitally deficient mice where the developmental consequences of eosinophil depletion are unknown. METHODS: We developed a novel conditional eosinophil-deficient strain of mice (iPHIL) through a gene knock-in strategy inserting the human diphtheria toxin (DT) receptor (DTR) into the endogenous eosinophil peroxidase genomic locus. RESULTS: Expression of DTR rendered resistant mouse eosinophil progenitors sensitive to DT without affecting any other cell types. The presence of eosinophils was shown to be unnecessary during the sensitization phase of either ovalbumin (OVA) or house dust mite (HDM) acute asthma models. However, eosinophil ablation during airway challenge led to a predominantly neutrophilic phenotype (>15% neutrophils) accompanied by allergen-induced histopathologies and airway hyper-responsiveness in response to methacholine indistinguishable from eosinophilic wild-type mice. Moreover, the iPHIL neutrophilic airway phenotype was shown to be a steroid-resistant allergic respiratory variant that was reversible upon the restoration of peripheral eosinophils. CONCLUSIONS: Eosinophil contributions to allergic immune/inflammatory responses appear to be limited to the airway challenge and not to the sensitization phase of allergen provocation models. The reversible steroid-resistant character of the iPHIL neutrophilic airway variant suggests underappreciated mechanisms by which eosinophils shape the character of allergic respiratory responses.


Asunto(s)
Eosinófilos/inmunología , Hipersensibilidad Respiratoria/inmunología , Alérgenos/inmunología , Animales , Asma/genética , Asma/inmunología , Asma/metabolismo , Citotoxicidad Inmunológica , Toxina Diftérica/administración & dosificación , Toxina Diftérica/inmunología , Modelos Animales de Enfermedad , Resistencia a Medicamentos , Eosinófilos/citología , Eosinófilos/efectos de los fármacos , Eosinófilos/metabolismo , Técnicas de Sustitución del Gen , Células Precursoras de Granulocitos/inmunología , Células Precursoras de Granulocitos/metabolismo , Factor de Crecimiento Similar a EGF de Unión a Heparina , Humanos , Péptidos y Proteínas de Señalización Intercelular/genética , Ratones , Ovalbúmina/inmunología , Fenotipo , Neumonía/genética , Neumonía/inmunología , Neumonía/metabolismo , Hipersensibilidad Respiratoria/genética , Hipersensibilidad Respiratoria/metabolismo , Hipersensibilidad Respiratoria/patología , Esteroides/farmacología , Células Th2/inmunología , Células Th2/metabolismo
5.
Allergy ; 68(9): 1177-84, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23931643

RESUMEN

BACKGROUND: Sputum eosinophilia has been shown to be a predictor of response to anti-eosinophil therapies in patients with airway diseases. However, quantitative cell counts and differentials of sputum are labor intensive. The objective of this study was to validate a novel ELISA-based assay of eosinophil peroxidase (EPX) in sputum as a rapid and reliable marker of airway eosinophils. METHODS: The utility of EPX-based ELISA as an eosinophil-specific assay was achieved through comparisons with sputum eosinophil differential counts in freshly prepared and archived patient samples from a variety of clinical settings. RESULTS: EPX levels in sputum correlated with eosinophil percentage (r(s) = 0.84) in asthma patients with varying degrees of airway eosinophilia. Significantly, unlike assays of other eosinophil granule proteins (e.g., ECP and EDN), which often detect the presence of these proteins even in asthma patients with neutrophilic bronchitis, EPX-based ELISA levels are not increased in this subset of asthma patients or in COPD patients lacking evidence of an airway eosinophilia. Moreover, sputum EPX was a surrogate marker of airway eosinophilia in other patient studies (e.g., allergen inhalation and treatment trials the anti-(IL-5) therapeutic Mepolizumab™). Finally, EPX levels in cytocentrifuged prepared sputum supernatants correlated with those from rapidly prepared noncentrifuged filtrates of sputum (r(s) = 0.94). CONCLUSION AND CLINICAL IMPLICATION: EPX-based ELISA is a valid, reliable, repeatable, and specific surrogate marker of eosinophils and/or eosinophil degranulation in the sputum of respiratory patients. The novel EPX assay is a valid and reproducible eosinophil-specific assay that can potentially be developed into a point-of-care assessment of eosinophil activity in airway secretions.


Asunto(s)
Peroxidasa del Eosinófilo/metabolismo , Eosinofilia/metabolismo , Enfermedades Respiratorias/metabolismo , Esputo/enzimología , Adulto , Anciano , Biomarcadores/metabolismo , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Eosinofilia/diagnóstico , Eosinofilia/fisiopatología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Enfermedades Respiratorias/diagnóstico , Enfermedades Respiratorias/fisiopatología
6.
Clin Exp Allergy ; 40(4): 563-75, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-20447076

RESUMEN

Discussions of eosinophils are often descriptions of end-stage effector cells with destructive capabilities mediated predominantly by released cytotoxic cationic granule proteins. Moreover, eosinophils in the medical literature are invariably associated with the pathologies linked with helminth infections or allergic diseases such as asthma. This has led to an almost fatalist view of eosinophil effector functions and associated therapeutic strategies targeting these cells that would make even William of Ockham proud - eosinophil effector functions have physiological consequences that increase patient morbidity/mortality and 'the only good eosinophils are dead eosinophils'. Unfortunately, the strengths of dogmas are also their greatest weaknesses. Namely, while the repetitive proclamation of dogmatic concepts by authoritative sources (i.e. reviews, meeting proceedings, textbooks, etc.) builds consensus within the medical community and lower the entropies surrounding difficult issues, they often ignore not easily explained details and place diminished importance on alternative hypotheses. The goal of this perspective is twofold: (i) we will review recent observations regarding eosinophils and their activities as well as reinterpret earlier data as part of the synthesis of a new paradigm. In this paradigm, we hypothesize that eosinophils accumulate at unique sites in response to cell turnover or in response to local stem cell activity(ies). We further suggest that this accumulation is part of one or more mechanisms regulating tissue homeostasis. Specifically, instead of immune cells exclusively mediating innate host defence, we suggest that accumulating tissue eosinophils are actually regulators of Local Immunity And/or Remodeling/Repair in both health and disease - the LIAR hypothesis; (ii) we want to be inflammatory (pun intended!) and challenge the currently common perspective of eosinophils as destructive end-stage effector cells. Our hope is to create more questions than we answer and provoke everyone to spend countless hours simply to prove us wrong!


Asunto(s)
Eosinófilos/inmunología , Eosinófilos/patología , Animales , Asma/inmunología , Asma/fisiopatología , Citotoxicidad Inmunológica , Eosinófilos/fisiología , Helmintiasis/inmunología , Helmintiasis/fisiopatología , Hematopoyesis , Humanos , Ratones , Neoplasias/inmunología , Neoplasias/fisiopatología
7.
Respir Med ; 104(9): 1254-62, 2010 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-20434897

RESUMEN

BACKGROUND: As it is unknown whether complete asthma remission or progression of asthma is associated with airway inflammation and remodeling, we assessed these characteristics in bronchial biopsies of relevant subsets of asthma patients. METHODS: Sputum and bronchial biopsies were obtained from asthma patients in remission (PC(20) histamine> 32 mg/ml, PC(20) AMP> 320 mg/ml) and from those with either a slow FEV(1) decline (< 30 ml/year) or fast decline (> 30 ml/year). Inflammatory cells and mediators were determined in sputum, inflammatory cells and aspects of airway remodeling in bronchial biopsies. RESULTS: Asthmatics in remission and asthma patients with a slow FEV(1) decline had a similar extent of airway inflammation and remodeling in sputum and bronchial biopsies. Asthma patients with a fast FEV(1) decline had high sputum eosinophil numbers. Moreover, FEV(1) decline (ml/year) correlated with sputum eosinophil numbers (Rs=0.51, p=0.003) and ECP levels (Rs=0.57, p=0.001). Airway remodeling, i.e. basement membrane thickness, correlated with sputum eosinophils (Rs=0.69, p<0.001), sputum ECP (Rs=0.46, p=0.018) and airway wall eosinophil numbers (Rs=0.49, p=0.002). CONCLUSIONS: Asthma, even when in remission, is accompanied by airway inflammation and remodeling. Data suggest that eosinophils are important in a subset of asthma patients by association to accelerated FEV(1) decline and change of basement membrane thickness.


Asunto(s)
Remodelación de las Vías Aéreas (Respiratorias)/fisiología , Asma/patología , Eosinofilia/patología , Adulto , Anciano , Asma/fisiopatología , Broncoscopía , Progresión de la Enfermedad , Eosinofilia/fisiopatología , Eosinófilos/patología , Femenino , Volumen Espiratorio Forzado/fisiología , Humanos , Mediadores de Inflamación/metabolismo , Masculino , Persona de Mediana Edad , Remisión Espontánea , Esputo/citología , Esputo/metabolismo
9.
Am J Physiol Lung Cell Mol Physiol ; 285(1): L114-20, 2003 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-12611815

RESUMEN

Airway function in health and disease as well as in response to bronchospastic stimuli (i.e., irritants, allergens, and inflammatory mediators) is controlled, in part, by cholinergic muscarinic receptor regulation of smooth muscle. In particular, the dependence of airway smooth muscle contraction/relaxation on heterotrimeric G protein-coupled receptor signaling suggests that these events underlie the responses regulating airway function. Galphaq-containing G proteins are proposed to be a prominent signaling pathway, and the availability of knockout mice deficient of this subunit has allowed for an investigation of its potential role in airway function. Airway responses in Galphaq-deficient mice (activities assessed by both tracheal tension and in vivo lung function measurements) were attenuated relative to wild-type controls. Moreover, ovalbumin sensitization/aerosol challenge of Galphaq-deficient mice also failed to elicit an allergen-induced increase in airway reactivity to methacholine. These findings indicate that cholinergic receptor-mediated responses are dependent on Galphaq-mediated signaling events and identify Galphaq as a potential target of preventative/intervening therapies for lung dysfunction.


Asunto(s)
Hiperreactividad Bronquial/fisiopatología , Broncoconstrictores/farmacología , Proteínas de Unión al GTP Heterotriméricas/genética , Proteínas de Unión al GTP Heterotriméricas/metabolismo , Cloruro de Metacolina/farmacología , Resistencia de las Vías Respiratorias/fisiología , Alérgenos/farmacología , Animales , Hiperreactividad Bronquial/inducido químicamente , Subunidades alfa de la Proteína de Unión al GTP Gq-G11 , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Liso/efectos de los fármacos , Músculo Liso/fisiopatología , Ovalbúmina/farmacología , Transducción de Señal/fisiología , Tráquea/efectos de los fármacos , Tráquea/fisiopatología
10.
Diabetes Obes Metab ; 4(3): 195-200, 2002 May.
Artículo en Inglés | MEDLINE | ID: mdl-12047398

RESUMEN

In vitro and in vivo studies have demonstrated that prostaglandins of the E series enhance muscle glucose uptake. We examined the effect of acute misoprostol (PGE1) administration on whole body insulin-mediated glucose disposal, as well as the major intracellular pathways of glucose metabolism in type 2 diabetic (n = 10) and non-diabetic (n = 4) subjects. Each subject received two 240-min euglycaemic insulin (40 mU/m2/min) clamp studies with tritiated glucose and indirect calorimetry. During one of the insulin clamp studies, 200 microg of misoprostol was ingested at 90 and 150 min after the start of the insulin infusion. Insulin-mediated total body glucose disposal, glycolysis, glycogenesis and glucose oxidation were similar during the insulin clamp studies performed without and with misoprostol in both the diabetic and non-diabetic groups. These results demonstrate that the acute administration of misoprostol does not enhance insulin-mediated glucose disposal in either type-2-diabetic or non-diabetic subjects.


Asunto(s)
Glucemia/metabolismo , Diabetes Mellitus Tipo 2/sangre , Misoprostol/farmacología , Glucemia/efectos de los fármacos , Índice de Masa Corporal , Colesterol/sangre , HDL-Colesterol/sangre , LDL-Colesterol/sangre , Femenino , Prueba de Tolerancia a la Glucosa , Hemoglobina Glucada/metabolismo , Humanos , Insulina/sangre , Masculino , Tasa de Depuración Metabólica , Persona de Mediana Edad , Misoprostol/sangre , Valores de Referencia , Triglicéridos/sangre
11.
Am J Physiol Lung Cell Mol Physiol ; 282(5): L1066-74, 2002 May.
Artículo en Inglés | MEDLINE | ID: mdl-11943672

RESUMEN

The potential role of airway interleukin-5 (IL-5) expression in eliciting mucus production was demonstrated in a pulmonary IL-5 transgenic mouse model (NJ.1726) in which naive transgenic mice display comparable levels of airway mucus relative to allergen-sensitized and -challenged wild-type mice. The intrinsic mucus accumulation of NJ.1726 was abolished in compound transgenic-gene knockout mice deficient of either CD4(+) cells [NJ.1726/CD4(-/-)] or alphabeta T cell receptor-positive (TCR(+)) cells [NJ.1726/alphabeta TCR(-/-)]. In addition, mucus production in naive NJ.1726 was inhibited by >90% after administration of the soluble anti-IL-4 receptor alpha-subunit antagonist. The loss of mucus production in NJ.1726/CD4(-/-), NJ.1726/alphabeta TCR(-/-), and anti-IL-4 receptor alpha-subunit antagonist-treated mice occurred notwithstanding the significant pulmonary eosinophilia and expansion of airway B cells induced by ectopic IL-5 expression. Furthermore, the loss of mucus accumulation occurred in these mice despite elevated levels of airway and peripheral IL-5, indicating that IL-5 does not directly induce goblet cell metaplasia and mucus production. Thus pulmonary expression of IL-5 alone is capable of inducing CD4(+) T cell-dependent goblet cell metaplasia, apparently mediated by IL-4 receptor alpha-subunit-ligand interactions, and represents a previously unrecognized novel pathway for augmenting allergen-induced mucus production.


Asunto(s)
Asma/patología , Linfocitos T CD4-Positivos/metabolismo , Interleucina-5/genética , Moco/metabolismo , Alérgenos/farmacología , Animales , Asma/inmunología , Asma/metabolismo , Eosinófilos/inmunología , Expresión Génica/inmunología , Células Caliciformes/metabolismo , Interleucina-5/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Moco/inmunología , Ovalbúmina/inmunología , Mucosa Respiratoria/metabolismo
12.
Am J Physiol Lung Cell Mol Physiol ; 282(1): L99-108, 2002 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-11741821

RESUMEN

CD4(+) T cells have a critical role in the development of allergic pulmonary inflammation, including the recruitment of eosinophils to the airway lumen and interstitium. The expression of interleukin (IL)-5 by CD4(+) cells has, in particular, often been lionized as the central link between allergic inflammation and the concomitant expansion or recruitment of eosinophils. The mechanism(s) by which CD4(+) T cells mediates eosinophil recruitment was assessed with gene knockout mice deficient for T cells or T cell subtypes and a unique IL-5 transgenic mouse (line NJ.1726) that constitutively overexpresses this cytokine in the lung epithelium. Pulmonary IL-5 expression is significantly attenuated in T cell- and CD4(+) but not CD8(+) cell-deficient animals, suggesting an obvious explanation for the lack of eosinophils in the lungs of T cell-deficient and CD4(-/-) mice. However, although the constitutive expression of IL-5 in the lung epithelium of NJ.1726 mice elicited an eosinophilia in the airway lumen of both naive and ovalbumin-treated mice, in the absence of CD4(+) cells, allergen-mediated eosinophil recruitment to the bronchoalveolar lavage fluid was abolished. Moreover, intranasal instillation of the potent eosinophil-specific chemokine eotaxin-2 was incapable of eliciting eosinophil recruitment in naive and ovalbumin-treated NJ.1726 CD4(-/-) mice, suggesting that eosinophil trafficking during allergic inflammatory responses is a consequence of a CD4(+) cell-mediated event(s) in addition to IL-5 expression and the establishment of a pulmonary chemokine gradient.


Asunto(s)
Linfocitos T CD4-Positivos/fisiología , Interleucina-5/metabolismo , Administración Intranasal , Alérgenos/inmunología , Animales , Quimiocina CCL24 , Quimiocinas CC/administración & dosificación , Quimiocinas CC/farmacología , Quimiotaxis de Leucocito/fisiología , Eosinófilos/efectos de los fármacos , Eosinófilos/patología , Eosinófilos/fisiología , Epitelio/metabolismo , Inmunización , Interleucina-5/genética , Pulmón/metabolismo , Pulmón/patología , Ratones , Ratones Noqueados , Ratones Transgénicos/genética , Ovalbúmina/inmunología , Eosinofilia Pulmonar/etiología , Eosinofilia Pulmonar/inmunología , Eosinofilia Pulmonar/patología
13.
J AOAC Int ; 84(5): 1393-406, 2001.
Artículo en Inglés | MEDLINE | ID: mdl-11601458

RESUMEN

Reliable monitoring technology is an essential component of effective regulation and risk management of environmental contaminants such as pesticides. Most environmental monitoring and analysis is currently conducted using instrumental techniques such as gas-liquid chromatography (GLC) and liquid chromatography (LC). Immunoanalysis provides powerful monitoring techniques that have emerged in the last 3 decades. This paper shows they can deliver rapid, accurate, and relatively inexpensive analysis with high throughput and that have the capability to be field oriented. The technique is versatile in application and can be formatted to suit different purposes such as quantitative analysis or simple "yes/no" tests that are field-portable. While there is a range of opinion on the merits of immunoassays as an analytical tool for pesticides, we suggest that this technology is best considered as complementary to GLC and LC, extending the range of capability for field monitoring. Supporting this view, an increasing number of successful applications of immunoassays to monitoring have been reported in recent years. We also report here the implications of recent developments in the field of immunodiagnostics and their application to monitoring of environmental contaminants. We emphasise that, together with adequate validation by instrumental techniques, immunoassays provide monitoring services yielding realistic and comprehensive data for risk management, allowing decisions on appropriate action by various authorities to be made.


Asunto(s)
Monitoreo del Ambiente/instrumentación , Inmunoquímica/instrumentación , Residuos de Plaguicidas/análisis , Plaguicidas/análisis , Animales , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoquímica/tendencias , Reproducibilidad de los Resultados
14.
Am J Physiol Lung Cell Mol Physiol ; 281(3): L653-9, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11504693

RESUMEN

Overexpression of interleukin (IL)-5 by the airway epithelium in mice using the rat CC10 promoter (NJ.1726 line) leads to several histopathologies characteristic of human asthma, including airway hyperreactivity (AHR). We investigated the contribution of B and T cells, as well as CD4 expression, to the development of AHR in IL-5 transgenic mice. NJ.1726 mice on a T cell or CD4 knockout background, but not on a B cell knockout background, lost intrinsic AHR. These effects occurred without decreases in IL-5 or eosinophils. We further investigated the contribution of alpha(4)-integrin signaling to the development of AHR in IL-5 transgenic mice through the administration of anti-CD49d (alpha(4)-integrin) antibody (PS/2). Administration of PS/2 resulted in immediate (16-h) inhibition of AHR. The inhibition of AHR was not associated with a decrease in airway eosinophils. These studies demonstrate that, despite the presence of increased levels of IL-5 and eosinophils in the lungs of NJ.1726 mice, CD4(+) cells and alpha(4)-integrin signaling are necessary for the intrinsic AHR that develops in IL-5 transgenic mice.


Asunto(s)
Antígenos CD/fisiología , Hiperreactividad Bronquial/etiología , Linfocitos T CD4-Positivos/fisiología , Interleucina-5/fisiología , Transducción de Señal/fisiología , Animales , Anticuerpos/farmacología , Antígenos CD/inmunología , Linfocitos B/citología , Bronquios/efectos de los fármacos , Bronquios/patología , Bronquios/fisiopatología , Hiperreactividad Bronquial/patología , Hiperreactividad Bronquial/fisiopatología , Líquido del Lavado Bronquioalveolar/química , Linfocitos T CD4-Positivos/patología , Eosinofilia/patología , Integrina alfa4 , Interleucina-5/genética , Pulmón/citología , Enfermedades Pulmonares/patología , Ratones , Ratones Transgénicos/genética
15.
J Immunol ; 167(3): 1672-82, 2001 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-11466391

RESUMEN

Paradigms of eosinophil effector function in the lungs of asthma patients invariably depend on activities mediated by cationic proteins released from secondary granules during a process collectively referred to as degranulation. In this study, we generated knockout mice deficient for eosinophil peroxidase (EPO) to assess the role(s) of this abundant secondary granule protein in an OVA-challenge model. The loss of EPO had no effect on the development of OVA-induced pathologies in the mouse. The absence of phenotypic consequences in these knockout animals extended beyond pulmonary histopathologies and airway changes, as EPO-deficient animals also displayed OVA-induced airway hyperresponsiveness after provocation with methacholine. In addition, EPO-mediated oxidative damage of proteins (e.g., bromination of tyrosine residues) recovered in bronchoalveolar lavage from OVA-treated wild-type mice was <10% of the levels observed in bronchoalveolar lavage recovered from asthma patients. These data demonstrate that EPO activities are inconsequential to the development of allergic pulmonary pathologies in the mouse and suggest that degranulation of eosinophils recruited to the lung in this model does not occur at levels comparable to those observed in humans with asthma.


Asunto(s)
Eosinófilos/enzimología , Eosinófilos/inmunología , Pulmón/metabolismo , Pulmón/patología , Ovalbúmina/inmunología , Peroxidasas/metabolismo , Proteínas/metabolismo , Alérgenos/administración & dosificación , Alérgenos/inmunología , Animales , Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/inmunología , Hiperreactividad Bronquial/patología , Degranulación de la Célula/inmunología , Movimiento Celular/genética , Movimiento Celular/inmunología , Cruzamientos Genéticos , Gránulos Citoplasmáticos/metabolismo , Gránulos Citoplasmáticos/ultraestructura , Modelos Animales de Enfermedad , Peroxidasa del Eosinófilo , Eosinófilos/metabolismo , Eosinófilos/ultraestructura , Inyecciones Intraperitoneales , Pulmón/enzimología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ovalbúmina/administración & dosificación , Oxidación-Reducción , Peroxidasas/deficiencia , Peroxidasas/genética , Hipersensibilidad Respiratoria/enzimología , Hipersensibilidad Respiratoria/genética , Hipersensibilidad Respiratoria/patología , Eliminación de Secuencia
16.
J Allergy Clin Immunol ; 107(6): 945-57, 2001 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11398070

RESUMEN

Etiologic discussions of allergic respiratory pathology frequently engender rabid constituencies of pro-T cell or proeosinophil disciples, each claiming, often with religious fervor, the importance of their leukocyte. However, increasing evidence suggests that the exclusionary rhetoric from either camp is inadequate to explain many of the pathologic changes occurring in the lung. Data from both asthmatic patient and mouse models of allergic respiratory inflammation suggest that, in addition to cell-autonomous activities, T-cell and eosinophil interactions may be critical to the onset and progression of pulmonary pathology. These studies also suggest that T-lymphocyte subpopulations and eosinophils communicate by means of both direct cell-cell interactions and through the secretion of inflammatory signals. Collectively, the data support an expanded view of T-cell and eosinophil activities in the lung, including both immunoregulative activities and downstream effector functions impinging directly on lung function.


Asunto(s)
Asma/inmunología , Eosinófilos/inmunología , Pulmón/inmunología , Hipersensibilidad Respiratoria/inmunología , Linfocitos T/inmunología , Animales , Asma/fisiopatología , Modelos Animales de Enfermedad , Humanos , Pulmón/citología , Ratones , Ratones Noqueados , Hipersensibilidad Respiratoria/fisiopatología
17.
Mamm Genome ; 12(5): 352-61, 2001 May.
Artículo en Inglés | MEDLINE | ID: mdl-11331942

RESUMEN

A unique family of ribonucleases was identified by exhaustive screening of genomic and cDNA libraries using a probe derived from a gene encoding a ribonuclease stored in the mouse eosinophil secondary granule. This family contains at least 13 genes, which encode ribonucleases, and two potential pseudogenes. The conserved sequence identity among these genes (approximately 70%), as well as the isolation/purification of these ribonucleases from eosinophil secondary granules, has led us to conclude that these genes form a unique clade in the mouse that we have identified as the Ear (Eosinophil-associated ribonuclease) gene family. Analyses of the nucleotide substitutions that have occurred among these ribonuclease genes reveal that duplication events within this family have been episodic, occurring within three unique periods during the past 18 x 10(6) years. Moreover, comparisons of non-synonymous (K(a)) vs. synonymous (K(s)) rates of nucleotide substitution show that although these genes conserve residues necessary for RNase activity, selective evolutionary pressure(s) exist such that acquired amino acid changes appear to be advantageous. The selective advantage of these amino acid changes is currently unclear, but the occurrence of this phenomenon in both the mouse and the human highlights the importance of these changes for Ear and, therefore, eosinophil effector function(s).


Asunto(s)
Eosinófilos/enzimología , Eosinófilos/metabolismo , Hematopoyesis/genética , Familia de Multigenes/genética , Ribonucleasas/genética , Secuencia de Aminoácidos , Animales , Northern Blotting , Evolución Molecular , Duplicación de Gen , Regulación del Desarrollo de la Expresión Génica , Humanos , Pulmón/enzimología , Pulmón/metabolismo , Ratones , Datos de Secuencia Molecular , Especificidad de Órganos , ARN Mensajero/análisis , ARN Mensajero/genética , Ribonucleasas/química , Homología de Secuencia de Aminoácido , Terminología como Asunto
18.
J Clin Invest ; 107(8): 949-59, 2001 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-11306598

RESUMEN

We have developed a transgenic mouse line, NJ.1638, which expresses high levels of IL-5 from T cells, with profound hematological consequences. Eosinophils comprise more than 60% of circulating white blood cells in these animals, with the total peripheral white blood cell counts increasing more than 40-fold relative to wild-type littermates. This extraordinary proliferative capacity is sustained by expanded sites of extramedullary hematopoiesis and is accompanied by multifocal, ectopic bone formation in the spleen. Histology of the splenic nodules revealed the presence of osteoid matrices and osteocytes trapped within mineralized trabecular plates. In addition, polarized light microscopy of calcified tissue sections revealed both woven bone and areas of organized lamellar bone. Morphometric assessments demonstrated that both the growth and mineralization of splenic bone occurred at rates nearly an order of magnitude higher than in skeletal bone. Skeletal bone metabolic parameters were also perturbed. We also observed heterotopic ossification of the spleen and perturbation of skeletal bone homeostasis following adoptive engraftment of transgenic marrow to wild-type recipients. These data suggest that IL-5 overexpression mediates bone formation through the mobilization of marrow-derived osteogenic progenitors and/or the inhibition of recruited osteoclasts.


Asunto(s)
Huesos/metabolismo , Interleucina-5/biosíntesis , Osificación Heterotópica/metabolismo , Bazo/patología , Animales , Trasplante de Médula Ósea , Calcificación Fisiológica , Calcio/metabolismo , Cartílago Articular/patología , Femenino , Expresión Génica , Hematopoyesis Extramedular , Interleucina-5/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Transgénicos , Microscopía de Polarización/métodos , Osteoblastos/metabolismo , Coloración y Etiquetado/métodos , Células Th2/metabolismo
19.
Am J Physiol Lung Cell Mol Physiol ; 280(4): L813-21, 2001 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-11238023

RESUMEN

Lymphocyte and/or eosinophil recruitment is dependent on the sequential interactions between adhesion molecules expressed on activated endothelial cells and both leukocyte subtypes. Endothelial P- and E-selectins mediate tethering and rolling of leukocytes through interactions with P-selectin glycoprotein ligand-1 (PSGL-1), and diapedesis subsequently occurs by engagement of endothelial vascular cell adhesion molecule-1 and CD49d (alpha(4)-integrins). The anti-inflammatory potential of interfering with these adhesive interactions was assessed with an ovalbumin challenge mouse model of asthma. Administration of a soluble form of PSGL-1 reduced eosinophils (80%) and lymphocytes (50%) in bronchoalveolar lavage fluid without affecting epithelial changes or airway hyperreactivity (AHR). In contrast, although administration of anti-CD49d monoclonal antibodies (PS/2) resulted in similar reductions in eosinophils (75%) and lymphocytes (50%), PS/2 reduced and abolished mucous cell metaplasia and AHR, respectively. Administration of both PSGL-1 and PS/2 had the additive effect of eliminating eosinophils from the airways (96% decrease), with few or no additional reductions (relative to PS/2 administration alone) in lymphocyte recruitment, mucous cell metaplasia, or AHR. These data show that eosinophils and lymphocytes differentially utilize adhesive interactions during recruitment and that the inhibition of AHR is independent of this recruitment.


Asunto(s)
Antígenos CD/fisiología , Hipersensibilidad/complicaciones , Leucocitos/efectos de los fármacos , Leucocitos/fisiología , Neumonía/prevención & control , Animales , Anticuerpos Monoclonales/farmacología , Antígenos CD/efectos de los fármacos , Antígenos CD/inmunología , Células Sanguíneas/patología , Hiperreactividad Bronquial/prevención & control , Combinación de Medicamentos , Eosinofilia/patología , Femenino , Integrina alfa4 , Pulmón/patología , Glicoproteínas de Membrana/farmacología , Ratones , Ratones Endogámicos BALB C , Neumonía/etiología , Neumonía/patología , Proteínas Recombinantes , Mucosa Respiratoria/efectos de los fármacos , Mucosa Respiratoria/patología , Solubilidad
20.
J Immunol ; 165(10): 5509-17, 2000 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-11067904

RESUMEN

The relationship between eosinophils and the development of Ag-induced pulmonary pathologies, including airway hyper-responsiveness, was investigated using mice deficient for the secondary granule component, major basic protein-1 (mMBP-1). The loss of mMBP-1 had no effect on OVA-induced airway histopathologies or inflammatory cell recruitment. Lung function measurements of knockout mice demonstrated a generalized hyporeactivity to methacholine-induced airflow changes (relative to wild type); however, this baseline phenotype was observable only with methacholine; no relative airflow changes were observed in response to another nonspecific stimulus (serotonin). Moreover, OVA sensitization/aerosol challenge of wild-type and mMBP-1(-/-) mice resulted in identical dose-response changes to either methacholine or serotonin. Thus, the airway hyper-responsiveness in murine models of asthma occurs in the absence of mMBP-1.


Asunto(s)
Alérgenos/inmunología , Asma/inmunología , Asma/patología , Proteínas Sanguíneas/fisiología , Eosinófilos/inmunología , Pulmón/inmunología , Pulmón/patología , Ribonucleasas , Alérgenos/administración & dosificación , Animales , Antígenos Helmínticos/administración & dosificación , Asma/genética , Proteínas Sanguíneas/biosíntesis , Proteínas Sanguíneas/deficiencia , Proteínas Sanguíneas/genética , Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/inmunología , Movimiento Celular/genética , Movimiento Celular/inmunología , Gránulos Citoplasmáticos/inmunología , Gránulos Citoplasmáticos/ultraestructura , Modelos Animales de Enfermedad , Proteínas en los Gránulos del Eosinófilo , Eosinófilos/patología , Eosinófilos/ultraestructura , Eliminación de Gen , Inyecciones Intraperitoneales , Mesocestoides/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Ratones Noqueados , Microscopía Electrónica , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología
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