RESUMEN
BACKGROUND: We previously demonstrated in mice that airway eosinophils traffic from the airway lumen into lung-draining paratracheal lymph nodes. However, mechanisms whereby eosinophils traverse from the lungs and home to paratracheal lymph nodes remain unclear. We investigated roles of cysteinyl leukotrienes in mediating eosinophil trafficking from lungs to paratracheal lymph nodes. METHODS: The expression of CCR7 was determined by flow cytometry. Transwell assays were used to test chemotactic responses of leukotriene C4 synthase-deficient and control airway eosinophils to the chemokine CCL19 ex vivo. Eosinophils from the spleens of IL-5 transgenic mice, fluorescently labeled ex vivo, were intratracheally injected into ovalbumin-sensitized and ovalbumin aerosol-challenged leukotriene C4 synthase-deficient and control mice. Eosinophils were identified by microscopy and flow cytometry in the lungs and paratracheal lymph nodes. RESULTS: Mouse eosinophils expressed CCR7, the receptor for CCL19, and responded chemotactically to CCL19. Leukotriene C4 synthase-deficient eosinophils exhibited impaired chemotaxis to CCL19 that was restored by exogenous leukotriene C4 . The migration of intratracheally injected eosinophils into paratracheal lymph nodes from distal alveolar lung was diminished in leukotriene C4 synthase-deficient mice compared with wild-type mice, with increased retention of eosinophils in the lungs of leukotriene C4 synthase-deficient mice. Exogenous administration of leukotriene C4 restored trafficking of eosinophils to paratracheal lymph nodes in leukotriene C4 synthase-deficient mice. CONCLUSIONS: Our findings that cysteinyl leukotrienes are involved in regulating airway and lung eosinophil migration into paratracheal lymph nodes identify previously unrecognized roles for the cysteinyl leukotrienes in regulating the pulmonary trafficking of eosinophils in experimental allergic asthma.
Asunto(s)
Quimiotaxis , Eosinófilos/citología , Leucotrieno C4/inmunología , Ganglios Linfáticos/citología , Animales , Asma/patología , Quimiocina CCL19/fisiología , Eosinófilos/metabolismo , Leucotrieno C4/administración & dosificación , Leucotrieno C4/deficiencia , Pulmón/citología , Ganglios Linfáticos/metabolismo , Ratones , Receptores CCR7/fisiologíaRESUMEN
A 32-year-old man with glutathione synthetase deficiency developing an acute metabolic crisis is described. During this acute episode, intracellular glutathione content in erythrocytes was below the detection limit (<0.3 mmol/L). Leukotriene C4 in CSF and urinary leukotriene E4 were massively decreased, indicating an imparied synthesis of cysteinyl leukotrienes. Clinical recovery after one week was accompanied by a clear improvement of these biochemical parameters. The highly disturbed glutathione synthesis is postulated to be the reason for a deficient synthesis of cysteinyl leukotrienes, which may at least in part be responsible for the severe clinical symptoms.
Asunto(s)
Glutatión Sintasa/deficiencia , Glutatión/deficiencia , Leucotrieno C4/líquido cefalorraquídeo , Errores Innatos del Metabolismo/metabolismo , Enfermedad Aguda , Adulto , Humanos , Leucotrieno C4/deficiencia , MasculinoRESUMEN
UNLABELLED: Leukotrienes (LTs) are a group of biologically highly active compounds which mainly include the cysteinyl leukotrienes LTC4, LTD4, LTE4 and LTB4. Biosynthesis is limited to a small number of different cells including brain tissue. Besides their well known function in the mediation of inflammation and host defence, cysteinyl LTs have neuromodulatory functions in the brain. Here we describe the recently discovered first two cases with a defect in the synthesis of LTs, LTC4 synthesis deficiency which is characterised by severe muscular hypotonia, psychomotor retardation, failure to thrive, microcephaly, and by the total absence of cysteinyl LTs in body fluids and deficient synthesis of the primary cysteinyl LT, LTC4, in blood cells. We describe the clinical and biochemical findings as well as the pathophysiological aspects of this condition and of further defects suggested in the synthetic pathway of LTs. Moreover, certain disease states which are known to be associated with secondary disturbances of LT degradation are also discussed. CONCLUSION: Leukotriene C4 synthesis deficiency represents a member of a newly recognised group of neurometabolic disorders which are probably underdiagnosed. Analysis of leukotrienes is recommended in all patients with neurological symptoms who have no apparently obvious metabolic cause.
Asunto(s)
Encefalopatías Metabólicas Innatas/diagnóstico , Encefalopatías Metabólicas Innatas/metabolismo , Leucotrieno C4/metabolismo , Encéfalo/enzimología , Encefalopatías Metabólicas Innatas/complicaciones , Encefalopatías Metabólicas Innatas/genética , Eicosanoides/metabolismo , Insuficiencia de Crecimiento/etiología , Glutatión/metabolismo , Humanos , Leucotrieno C4/biosíntesis , Leucotrieno C4/deficiencia , Proteínas de la Membrana/metabolismo , Microcefalia/etiología , Hipotonía Muscular/etiología , Trastornos Psicomotores/etiología , gamma-Glutamiltransferasa/metabolismoRESUMEN
To characterize the role of GM-CSF in pulmonary fibrosis, we have studied bleomycin-induced fibrosis in wild-type mice vs mice with a targeted deletion of the GM-CSF gene (GM-CSF-/- mice). Without GM-CSF, pulmonary fibrosis was worse both histologically and quantitatively. These changes were not related to enhanced recruitment of inflammatory cells because wild-type and GM-CSF-/- mice recruited equivalent numbers of cells to the lung following bleomycin. Interestingly, recruitment of eosinophils was absent in GM-CSF-/- mice. We investigated whether the enhanced fibrotic response in GM-CSF-/- animals was due to a deficiency in an endogenous down-regulator of fibrogenesis. Analysis of whole lung homogenates from saline- or bleomycin-treated mice revealed that GM-CSF-/- animals had reduced levels of PGE2. Additionally, alveolar macrophages were harvested from wild-type and GM-CSF-/- mice that had been exposed to bleomycin. Although bleomycin treatment impaired the ability of alveolar macrophages from wild-type mice to synthesize PGE2, alveolar macrophages from GM-CSF-/- mice exhibited a significantly greater defect in PGE2 synthesis than did wild-type cells. Exogenous addition of GM-CSF to alveolar macrophages reversed the PGE2 synthesis defect in vitro. Administration of the PG synthesis inhibitor, indomethacin, to wild-type mice during the fibrogenic phase postbleomycin worsened the severity of fibrosis, implying a causal role for PGE2 deficiency in the evolution of the fibrotic lesion. These data demonstrate that GM-CSF deficiency results in enhanced fibrogenesis in bleomycin-induced pulmonary fibrosis and indicate that one mechanism for this effect is impaired production of the potent antifibrotic eicosanoid, PGE2.
Asunto(s)
Bleomicina/farmacología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/fisiología , Prostaglandinas/fisiología , Fibrosis Pulmonar/etiología , Fibrosis Pulmonar/metabolismo , Animales , Bleomicina/administración & dosificación , División Celular/genética , Dinoprostona/antagonistas & inhibidores , Dinoprostona/biosíntesis , Dinoprostona/deficiencia , Esquema de Medicación , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Factor Estimulante de Colonias de Granulocitos y Macrófagos/deficiencia , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Indometacina/administración & dosificación , Inyecciones Intramusculares , Inyecciones Intraperitoneales , Intubación Intratraqueal , Cinética , Recuento de Leucocitos , Leucotrieno C4/biosíntesis , Leucotrieno C4/deficiencia , Metabolismo de los Lípidos , Lípidos/biosíntesis , Macrófagos Alveolares/metabolismo , Macrófagos Alveolares/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Monocitos/patología , Fibrosis Pulmonar/inducido químicamente , Fibrosis Pulmonar/genética , Cloruro de Sodio/administración & dosificaciónAsunto(s)
Leucotrieno C4/deficiencia , Errores Innatos del Metabolismo/enzimología , gamma-Glutamiltransferasa/deficiencia , Células Cultivadas , Resultado Fatal , Femenino , Glutatión Transferasa/metabolismo , Humanos , Lactante , Leucotrieno C4/sangre , Leucotrieno C4/líquido cefalorraquídeo , Leucotrieno C4/orina , Leucotrieno D4/líquido cefalorraquídeo , Leucotrieno E4/líquido cefalorraquídeo , Errores Innatos del Metabolismo/sangre , Errores Innatos del Metabolismo/líquido cefalorraquídeo , Errores Innatos del Metabolismo/fisiopatología , Monocitos/citología , Monocitos/enzimologíaRESUMEN
Cysteinyl leukotrienes (LTC(4), LTD(4), LTE(4)) are potent lipid mediators derived from arachidonate in the 5-lipoxygenase pathway. Recently, the first inborn error of leukotriene synthesis, LTC(4)-synthesis deficiency, has been identified in association with a fatal developmental syndrome. The absence of leukotrienes in cerebrospinal fluid was one of the most striking biochemical findings in this disorder. We analysed leukotrienes in cerebrospinal fluid of patients with a broad spectrum of other well-defined inborn errors of metabolism, including glutathione synthetase deficiency (n=2), Zellweger syndrome (n=3), mitochondrial disorders (n=8), fatty acid oxidation defects (n=7), organic acidurias (n=7), neurotransmitter defects (n=5) and patients with non-specific neurological symptoms, as a reference population (n=120). The concentrations of leukotrienes were not related to age. Representative percentiles were calculated as reference intervals of each leukotriene. In all patients with an inborn error of metabolism concentration of cysteinyl leukotrienes and LTB(4) did not differ from the reference group. Our results indicate that absence of cysteinyl leukotrienes (<5 pg/ml) in association with normal or increased LTB(4) (50.0-67.3 pg/ml) is pathognomonic for LTC(4)-synthesis deficiency. The unique profile of leukotrienes in cerebrospinal fluid in this new disorder is primarily related to the defect and represents a new diagnostic approach.
Asunto(s)
Leucotrieno C4/líquido cefalorraquídeo , Leucotrieno C4/deficiencia , Leucotrienos/líquido cefalorraquídeo , Errores Innatos del Metabolismo/líquido cefalorraquídeo , Adolescente , Estudios de Casos y Controles , Niño , Preescolar , Humanos , Lactante , Recién Nacido , Leucotrieno B4/líquido cefalorraquídeo , Leucotrieno D4/líquido cefalorraquídeo , Leucotrieno E4/líquido cefalorraquídeo , Errores Innatos del Metabolismo/diagnóstico , Valores de ReferenciaRESUMEN
Leukotrienes are potent oxygenated metabolites derived from the 5-lipoxygenase pathway of arachidonic acid metabolism. They comprise the cysteinyl leukotrienes (LTC4, LTD4, LTE4) and LTB4. The rate limiting step in the formation of cysteinyl leukotrienes is the conversion of LTA4 to LTC4 catalyzed by the enzyme LTC4 synthase. Recently, the first inborn error of leukotriene synthesis, LTC4-synthesis deficiency, has been identified in a patient with a fatal developmental syndrome. We report on an additional infant presenting with severe muscular hypotonia, symmetrical extension in the lower extremities and psychomotor retardation who died at the age of 6 months. Despite intensive investigations no specific diagnosis could be made. Leukotrienes were subsequently analyzed in the cerebrospinal fluid. Concentrations of LTC4, LTD4 and LTE4 were below the detection limit (< 5 pg/ml) whereas LTB4 was found to be in the upper normal range. The absence of cysteinyl leukotrienes with normal LTB4 concentration in cerebrospinal fluid is unique and seems to be pathognomonic for LTC4-synthesis deficiency. Our patient most likely represents the second case described so far with this condition. This report provides further evidence that LTC4-synthesis deficiency represents a new neurometabolic disorder.
Asunto(s)
Leucotrieno C4/líquido cefalorraquídeo , Leucotrieno C4/deficiencia , Errores Innatos del Metabolismo/diagnóstico , Resultado Fatal , Femenino , Humanos , Lactante , Leucotrieno C4/biosíntesis , Errores Innatos del Metabolismo/líquido cefalorraquídeo , Errores Innatos del Metabolismo/complicaciones , Hipotonía Muscular/etiología , Trastornos Psicomotores/etiologíaRESUMEN
BACKGROUND: Cysteinyl leukotrienes (LTC4, LTD4, LTE4) are potent lipid mediators derived from arachidonic acid in the 5-lipoxygenase pathway that exert profound biological effects. We investigated synthesis and metabolism of leukotrienes in an infant who presented with muscular hypotonia, psychomotor retardation, failure to thrive, and microcephaly. The course of the disease was rapidly progressive and the infant died aged 6 months. METHODS: Cysteinyl leukotrienes and LTB4 were analysed in cerebrospinal fluid, plasma, urine, and stimulated monocytes by EIA. We measured [3H]-LTC4 formation from [3H]-LTA4 in monocytes and platelets by radio-high-pressure liquid chromatography. FINDINGS: Concentrations of LTC4 and its metabolites were below the detection limit in the cerebrospinal fluid, plasma and urine. LTC4 could not be generated in stimulated monocytes, whereas LTB4 synthesis was increased. [3H]-LTC4 could not be made from [3H]-LTA4 in the patient's monocytes or platelets. INTERPRETATION: In this patient, inability to synthesise LTC4 suggests a deficiency of LTC4 synthase. This defect is a new inborn error of human eicosanoid metabolism and may be associated with the clinical disorder. Leukotriene analysis should be done in all patients with neurological symptoms who are candidates for metabolic diseases.
Asunto(s)
Anomalías Múltiples/metabolismo , Glutatión Transferasa/deficiencia , Leucotrieno C4/deficiencia , Errores Innatos del Metabolismo/metabolismo , Anomalías Múltiples/genética , Estudios de Casos y Controles , Consanguinidad , Insuficiencia de Crecimiento/genética , Femenino , Humanos , Recién Nacido , Leucotrieno C4/biosíntesis , Leucotrieno C4/fisiología , Errores Innatos del Metabolismo/genética , Microcefalia/genética , Hipotonía Muscular/genética , Trastornos Psicomotores/genéticaRESUMEN
Leukotrienes (LT) are inflammatory mediators which can also exert regulatory effects on human myelopoiesis. We have studied the LT-producing capacity of freshly isolated leucocyte suspensions (containing blast cells in variable proportions) from 41 patients with acute myeloid leukaemia (AML) or chronic myeloid leukaemia (CML) in blast crisis (CMLbc) at diagnosis or relapse/resistant disease. Leucocyte suspensions from 19/29 AML patients (66%), and 2/12 CMLbc patients (17%; P = 0.012) demonstrated deficient capacity to synthesize LT from endogenous substrate after ionophore A23187 stimulation. Thus, these cells produced < 8 pmol LTB4+LTC4/10(6) cells (< 20% of mean LT formation in leucocyte suspensions from 18 healthy subjects). Addition of exogenous arachidonic acid did not normalize the LT synthesis in poor-producing cell suspensions. Purified, morphologically mature granulocytes from two AML patients also failed to produce normal amounts of LT. In leucocyte suspensions from the remaining 20 AML/CMLbc patients A23187 provoked LT biosynthesis, with markedly increased production of LTC4, but decreased LTB4 formation. Furthermore, elevated conversion of exogenous LTA4 to LTC4 was noted in the patient samples, independent of their capacity to produce LT after A23187 stimulation. The percentage of blast cells in patient white blood cell differential counts correlated inversely with ionophore-induced LT synthesis, but positively with the conversion of exogenous LTA4 to LTC4. The results suggest elevated LTC4 synthase activity and suppressed 5-lipoxygenase activity as novel enzymatic features of myeloid leukaemia patients with immature phenotype.