RESUMEN
Allergic airway diseases are immune disorders associated with heightened type 2 immune responses and IL-5 and IL-13 production at the site of inflammation. We have previously reported that cyclooxygenase (COX) inhibition by indomethacin augmented allergic airway inflammation in a STAT6-independent manner. However, the key COX product(s) responsible for restraining indomethacin-mediated STAT6-independent allergic inflammation is unknown. In this study, using the mouse model of OVA-induced allergic airway inflammation, we identified that PGI2 receptor (IP) signaling was critical for indomethacin-induced, STAT6-independent proallergic effects. We demonstrated that IP deficiency increased inflammatory cell infiltration, eosinophilia, and IL-5 and IL-13 expression in the lung in a STAT6-independent manner. The augmented STAT6-independent allergic inflammation correlated with enhanced primary immune responses to allergic sensitization and elevated production of multiple inflammatory chemokines (CCL11, CCL17, CCL22, and CXCL12) in the lung after allergen challenge. We also showed that the PGI2 analogue cicaprost inhibited CD4 T cell proliferation and IL-5 and IL-13 expression in vitro, and IP deficiency diminished the stimulatory effect of indomethacin on STAT6-independent IL-5 and IL-13 responses in vivo. The inhibitory effects of PGI2 and the IP signaling pathway on CD4 T cell activation, inflammatory chemokine production, and allergic sensitization and airway inflammation suggest that PGI2 and its analogue iloprost, both Food and Drug Administration-approved drugs, may be useful in treating allergic diseases and asthma. In addition, inhibiting PGI2 signaling by drugs that either block PGI2 production or restrain IP signaling may augment STAT6-independent pathways of allergic inflammation.
Asunto(s)
Alérgenos/inmunología , Pulmón/inmunología , Activación de Linfocitos/efectos de los fármacos , Receptores de Epoprostenol/metabolismo , Factor de Transcripción STAT6/metabolismo , Alérgenos/administración & dosificación , Animales , Antihipertensivos/farmacología , Asma/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/fisiología , Proliferación Celular , Quimiocinas/biosíntesis , Quimiocinas/inmunología , Epoprostenol/administración & dosificación , Epoprostenol/análogos & derivados , Epoprostenol/farmacología , Hipersensibilidad , Indometacina , Inflamación , Interleucina-13/genética , Interleucina-13/inmunología , Interleucina-5/genética , Interleucina-5/inmunología , Pulmón/fisiopatología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Ovalbúmina/inmunología , Receptores de Epoprostenol/deficiencia , Receptores de Epoprostenol/genética , Factor de Transcripción STAT6/deficiencia , Factor de Transcripción STAT6/genética , Factor de Transcripción STAT6/inmunología , Transducción de Señal , Células Th2/inmunologíaRESUMEN
Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, a multienzyme complex, is the major source for production of reactive oxygen species (ROS). ROS are increased in allergic diseases, such as asthma, but the role of ROS in disease pathogenesis remains uncertain. We hypothesized that mice unable to generate ROS via the NADPH oxidase pathway would have decreased allergic airway inflammation. To test this hypothesis, we studied gp91phox(-/-) mice in a model of allergic airway inflammation after sensitization and challenge with ovalbumin. Serum, bronchoalveolar lavage fluid, and lungs were then examined for evidence of allergic inflammation. We found that mice lacking a functional NADPH oxidase complex had significantly decreased ROS production and allergic airway inflammation, compared with wild-type (WT) control animals. To determine the mechanism by which allergic inflammation was inhibited by gp91phox deficiency, we cultured bone marrow-derived dendritic cells from WT and gp91phox(-/-) mice and activated them with LPS. IL-12 expression was significantly increased in the gp91phox(-/-) bone marrow-derived dendritic cells, suggesting that the cytokine profile produced in the absence of gp91phox enhanced the conditions leading to T helper (Th) type 1 differentiation, while inhibiting Th2 polarization. Splenocytes from sensitized gp91phox(-/-) animals produced significantly less IL-13 in response to ovalbumin challenge in vitro compared with splenocytes from sensitized WT mice, suggesting that NADPH oxidase promotes allergic sensitization. In contrast, inflammatory cytokines produced by T cells cultured from WT and gp91phox(-/-) mice under Th0, Th1, Th2, and Th17 conditions were not significantly different. This study demonstrates the importance of NADPH oxidase activity and ROS production in a murine model of asthma.
Asunto(s)
Asma/inmunología , Interleucina-12/inmunología , Interleucina-13/inmunología , Pulmón/inmunología , Glicoproteínas de Membrana/inmunología , NADPH Oxidasas/inmunología , Especies Reactivas de Oxígeno/inmunología , Animales , Asma/inducido químicamente , Asma/genética , Asma/patología , Líquido del Lavado Bronquioalveolar/química , Células Cultivadas , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Células Dendríticas/patología , Femenino , Eliminación de Gen , Interleucina-12/biosíntesis , Interleucina-13/biosíntesis , Lipopolisacáridos/farmacología , Pulmón/metabolismo , Pulmón/patología , Glicoproteínas de Membrana/deficiencia , Glicoproteínas de Membrana/genética , Ratones , Ratones Noqueados , NADPH Oxidasa 2 , NADPH Oxidasas/deficiencia , NADPH Oxidasas/genética , Ovalbúmina/inmunología , Ovalbúmina/farmacología , Especies Reactivas de Oxígeno/metabolismo , Células TH1/inmunología , Células TH1/patología , Balance Th1 - Th2 , Células Th17/inmunología , Células Th17/patología , Células Th2/inmunología , Células Th2/patologíaRESUMEN
Suppressor of cytokine signaling (SOCS) 3 attenuates proinflammatory signaling mediated by the signal transducer and activator of transcription (STAT) family of proteins. But acute inflammation can occur after exposure to pathogen-derived inducers staphylococcal enterotoxin B (SEB) and lipopolysaccharide (LPS), or the lectin concanavalin A (ConA), suggesting that physiologic levels of SOCS3 are insufficient to stem proinflammatory signaling under pathogenic circumstances. To test this hypothesis, we developed recombinant cell-penetrating forms of SOCS3 (CP-SOCS3) for intracellular delivery to counteract SEB-, LPS- and ConA-induced inflammation. We found that CP-SOCS3 was distributed in multiple organs within 2 h and persisted for at least 8 h in leukocytes and lymphocytes. CP-SOCS3 protected animals from lethal effects of SEB and LPS by reducing production of inflammatory cytokines and attenuating liver apoptosis and hemorrhagic necrosis. It also reduced ConA-induced liver apoptosis. Thus, replenishing the intracellular stores of SOCS3 with CP-SOCS3 effectively suppresses the devastating effects of acute inflammation.
Asunto(s)
Apoptosis/efectos de los fármacos , Enterotoxinas/toxicidad , Inflamación/tratamiento farmacológico , Proteínas Recombinantes/uso terapéutico , Transducción de Señal/efectos de los fármacos , Infecciones Estafilocócicas/complicaciones , Proteínas Supresoras de la Señalización de Citocinas/uso terapéutico , Animales , Concanavalina A/toxicidad , Citocinas/sangre , Inflamación/etiología , Leucocitos/metabolismo , Lipopolisacáridos/toxicidad , Hígado/patología , Linfocitos/metabolismo , Ratones , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacocinética , Proteínas Recombinantes/farmacología , Factor de Transcripción STAT1/metabolismo , Infecciones Estafilocócicas/metabolismo , Proteína 3 Supresora de la Señalización de Citocinas , Proteínas Supresoras de la Señalización de Citocinas/metabolismo , Proteínas Supresoras de la Señalización de Citocinas/farmacocinética , Proteínas Supresoras de la Señalización de Citocinas/farmacologíaRESUMEN
The mortality rate for acute lung injury (ALI) is reported to be between 35-40%, and there are very few treatment strategies that improve the death rate from this condition. Previous studies have suggested that signaling through the prostaglandin (PG) I(2) receptor may protect against bleomycin-induced ALI in mice. We found that mice that overexpress PGI synthase (PGIS) in the airway epithelium were significantly protected against bleomycin-induced mortality and had reduced parenchymal consolidation, apoptosis of lung tissue, and generation of F(2)-isoprostanes compared with littermate wild-type controls. In addition, we show for the first time in both in vivo and in vitro experiments that PGI(2) induced the expression of NADP (H): quinoneoxidoreductase 1 (Nqo 1), an enzyme that prevents the generation of reactive oxygen species. PGI(2) induction of Nqo 1 provides a possible novel mechanism by which this prostanoid protects against bleomycin-induced mortality and identifies a potential therapeutic target for human ALI.
Asunto(s)
Lesión Pulmonar Aguda/genética , Lesión Pulmonar Aguda/prevención & control , Epoprostenol , Pulmón/metabolismo , NAD(P)H Deshidrogenasa (Quinona) , Prostaglandina-Endoperóxido Sintasas , Mucosa Respiratoria/metabolismo , Transducción de Señal , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/metabolismo , Lesión Pulmonar Aguda/mortalidad , Animales , Apoptosis/genética , Bleomicina/efectos adversos , Líquido del Lavado Bronquioalveolar/química , Epoprostenol/biosíntesis , F2-Isoprostanos/análisis , F2-Isoprostanos/biosíntesis , Femenino , Cromatografía de Gases y Espectrometría de Masas , Expresión Génica , Humanos , Inmunohistoquímica , Pulmón/efectos de los fármacos , Pulmón/patología , Ratones , Ratones Transgénicos , NAD(P)H Deshidrogenasa (Quinona)/biosíntesis , NAD(P)H Deshidrogenasa (Quinona)/genética , Reacción en Cadena de la Polimerasa , Prostaglandina-Endoperóxido Sintasas/biosíntesis , Prostaglandina-Endoperóxido Sintasas/genética , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Especies Reactivas de Oxígeno/metabolismo , Receptores de Epoprostenol/metabolismo , Pruebas de Función Respiratoria , Mucosa Respiratoria/efectos de los fármacos , Mucosa Respiratoria/patología , Tasa de SupervivenciaRESUMEN
IL-4 contributes to immunopathology induced in mice by primary respiratory syncytial virus (RSV) infection. However, the cellular source of IL-4 in RSV infection is unknown. We identified CD3(-)CD49b(+) cells as the predominant source of IL-4 in the lungs of RSV-infected BALB/c mice. We ruled out T cells, NK cells, NKT cells, mast cells, and eosinophils as IL-4 expressors in RSV infection by flow cytometry. Using IL4 GFP reporter mice (4get) mice, we identified the IL-4-expressing cells in RSV infection as basophils (CD3(-)CD49b(+)FcepsilonRI(+)c-kit(-)). Because STAT1(-/-) mice have an enhanced Th2-type response to RSV infection, we also sought to determine the cellular source and role of IL-4 in RSV-infected STAT1(-/-) mice. RSV infection resulted in significantly more IL-4-expressing CD3(-)CD49b(+) cells in the lungs of STAT1(-/-) mice than in BALB/c mice. CD49b(+)IL-4(+) cells sorted from the lungs of RSV-infected STAT1(-/-) mice and stained with Wright-Giemsa had basophil characteristics. As in wild-type BALB/c mice, IL-4 contributed to lung histopathology in RSV-infected STAT1(-/-) mice. Depletion of basophils in RSV-infected STAT1(-/-) mice reduced lung IL-4 expression. Thus, we show for the first time that a respiratory virus (RSV) induced basophil accumulation in vivo. Basophils were the primary source of IL-4 in the lung in RSV infection, and STAT1 was a negative regulator of virus-induced basophil IL-4 expression.
Asunto(s)
Basófilos/virología , Regulación de la Expresión Génica , Interleucina-4/genética , Pulmón/metabolismo , Infecciones por Virus Sincitial Respiratorio/inmunología , Factor de Transcripción STAT1/fisiología , Animales , Basófilos/metabolismo , Basófilos/patología , Citometría de Flujo , Inmunofenotipificación , Pulmón/patología , Ratones , Ratones Endogámicos BALB CRESUMEN
This article describes how Dr Ernest Goodpasture's 1919 study of influenza became the basis for his name being attached to pulmonary-renal syndromes by Stanton and Tange in 1958. Dr Goodpasture was an unwilling participant in this naming, and in retrospect, the patient he described who served as the prototypic case probably had a vasculitic syndrome. Finally, Dr Goodpasture's major contributions to virology are summarized.
Asunto(s)
Enfermedad por Anticuerpos Antimembrana Basal Glomerular/historia , Gripe Humana/historia , Patología/historia , Terminología como Asunto , Historia del Siglo XX , Humanos , Virología/historiaRESUMEN
Histoplasmosis was proven to be a fungal infection 70 years ago by Dr William DeMonbreun, at the time an assistant professor in the Department of Pathology at Vanderbilt Medical School. The significance of his work is analyzed in relationship to the evolution of knowledge about this important fungal infection. His discovery was also central to establishing the legitimacy of the recently reorganized medical school. Vanderbilt Medical School in 1925 was an experiment in building an educational institution essentially from scratch-the outcome of the experiment could be judged in the near term only by research productivity and Dr DeMonbreun's work was one of the 5 major discoveries made at Vanderbilt in the first decade of its existence. Further, his work is the bedrock on which Christie and Peterson later showed that histoplasmosis was endemic in the Ohio River Valley. Their studies plus a host of case reports and reviews up to recent times have contributed significantly to the academic standing of Vanderbilt. Heretofore unpublished illustrations and details about the prototypic cases are included for historical purposes. New light is also shed on the chain of circumstances that led to Vanderbilt's role in the evolution of knowledge about histoplasmosis. Finally, information is provided about Dr DeMonbreun's career after his discovery.
Asunto(s)
Histoplasmosis/patología , Patología/historia , Animales , Histoplasmosis/historia , Histoplasmosis/veterinaria , Historia del Siglo XX , Humanos , Estados UnidosRESUMEN
The paper is in three parts. 1) A description of acute anterior poliomyelitis; 2) A summary of the condition of post-polio syndrome; 3) A description of two small epidemics of poliomyelitis juxtaposed and related to one another.
Asunto(s)
Poliomielitis/historia , Síndrome Pospoliomielitis/historia , Distinciones y Premios , Brotes de Enfermedades/historia , Historia del Siglo XX , Humanos , Poliomielitis/epidemiología , Poliomielitis/patología , Síndrome Pospoliomielitis/epidemiología , Síndrome Pospoliomielitis/patología , Sociedades Médicas , Estados Unidos/epidemiologíaRESUMEN
BACKGROUND: Elevated cholesterol and triglycerides in blood lead to atherosclerosis and fatty liver, contributing to rising cardiovascular and hepatobiliary morbidity and mortality worldwide. METHODS AND RESULTS: A cell-penetrating nuclear transport modifier (NTM) reduced hyperlipidemia, atherosclerosis, and fatty liver in low-density lipoprotein receptor-deficient mice fed a Western diet. NTM treatment led to lower cholesterol and triglyceride levels in blood compared with control animals (36% and 53%, respectively; P<0.005) and liver (41% and 34%, respectively; P<0.05) after 8 weeks. Atherosclerosis was reduced by 63% (P<0.0005), and liver function improved compared with saline-treated controls. In addition, fasting blood glucose levels were reduced from 209 to 138 mg/dL (P<0.005), and body weight gain was ameliorated (P<0.005) in NTM-treated mice, although food intake remained the same as that in control animals. The NTM used in this study, cSN50.1 peptide, is known to modulate nuclear transport of stress-responsive transcription factors such as nuclear factor kappa B, the master regulator of inflammation. This NTM has now been demonstrated to also modulate nuclear transport of sterol regulatory element-binding protein (SREBP) transcription factors, the master regulators of cholesterol, triglyceride, and fatty acid synthesis. NTM-modulated translocation of SREBPs to the nucleus was associated with attenuated transactivation of their cognate genes that contribute to hyperlipidemia. CONCLUSIONS: Two-pronged control of inflammation and dyslipidemia by modulating nuclear transport of their critical regulators offers a new approach to comprehensive amelioration of hyperlipidemia, atherosclerosis, fatty liver, and their potential complications.
Asunto(s)
Aterosclerosis/tratamiento farmacológico , Núcleo Celular/metabolismo , Péptidos de Penetración Celular/uso terapéutico , Hígado Graso/tratamiento farmacológico , Hipercolesterolemia/tratamiento farmacológico , FN-kappa B/metabolismo , Péptidos/uso terapéutico , Proteínas de Unión a los Elementos Reguladores de Esteroles/metabolismo , Transporte Activo de Núcleo Celular/efectos de los fármacos , Animales , Aterosclerosis/metabolismo , Núcleo Celular/efectos de los fármacos , Péptidos de Penetración Celular/farmacología , Colesterol/metabolismo , Grasas de la Dieta/metabolismo , Modelos Animales de Enfermedad , Hígado Graso/metabolismo , Femenino , Hipercolesterolemia/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Ratones , Ratones Transgénicos , FN-kappa B/efectos de los fármacos , Péptidos/farmacología , Proteínas de Unión a los Elementos Reguladores de Esteroles/efectos de los fármacos , Factores de Transcripción/efectos de los fármacos , Factores de Transcripción/metabolismo , Triglicéridos/metabolismoRESUMEN
BACKGROUND: In the last ten years, bioterrorism has become a serious threat and challenge to public health worldwide. Pulmonary anthrax caused by airborne Bacillus anthracis spores is a life-threatening disease often refractory to antimicrobial therapy. Inhaled spores germinate into vegetative forms that elaborate an anti-phagocytic capsule along with potent exotoxins which disrupt the signaling pathways governing the innate and adaptive immune responses and cause endothelial cell dysfunction leading to vascular injury in the lung, hypoxia, hemorrhage, and death. METHODS/PRINCIPAL FINDINGS: Using a murine model of pulmonary anthrax disease, we showed that a nuclear transport modifier restored markers of the innate immune response in spore-infected animals. An 8-day protocol of single-dose ciprofloxacin had no significant effect on mortality (4% survival) of A/J mice lethally infected with B. anthracis Sterne. Strikingly, mice were much more likely to survive infection (52% survival) when treated with ciprofloxacin and a cell-penetrating peptide modifier of host nuclear transport, termed cSN50. In B. anthracis-infected animals treated with antibiotic alone, we detected a muted innate immune response manifested by cytokines, tumor necrosis factor alpha (TNFα), interleukin (IL)-6, and chemokine monocyte chemoattractant protein-1 (MCP-1), while the hypoxia biomarker, erythropoietin (EPO), was greatly elevated. In contrast, cSN50-treated mice receiving ciprofloxacin demonstrated a restored innate immune responsiveness and reduced EPO level. Consistent with this improvement of innate immunity response and suppression of hypoxia biomarker, surviving mice in the combination treatment group displayed minimal histopathologic signs of vascular injury and a marked reduction of anthrax bacilli in the lungs. CONCLUSIONS: We demonstrate, for the first time, that regulating nuclear transport with a cell-penetrating modifier provides a cytoprotective effect, which enables the host's immune system to reduce its susceptibility to lethal B. anthracis infection. Thus, by combining a nuclear transport modifier with antimicrobial therapy we offer a novel adjunctive measure to control florid pulmonary anthrax disease.
Asunto(s)
Carbunco/tratamiento farmacológico , Carbunco/mortalidad , Antiinfecciosos/administración & dosificación , Enfermedades Pulmonares/tratamiento farmacológico , Enfermedades Pulmonares/mortalidad , Transporte Activo de Núcleo Celular/efectos de los fármacos , Animales , Carbunco/complicaciones , Carbunco/patología , Antiinfecciosos/farmacología , Péptidos de Penetración Celular/administración & dosificación , Péptidos de Penetración Celular/farmacología , Ciprofloxacina/administración & dosificación , Ciprofloxacina/farmacología , Citocinas/sangre , Citocinas/metabolismo , Modelos Animales de Enfermedad , Combinación de Medicamentos , Femenino , Enfermedades Pulmonares/etiología , Enfermedades Pulmonares/patología , Proteínas de la Membrana/administración & dosificación , Proteínas de la Membrana/farmacología , Ratones , Péptidos Cíclicos/administración & dosificación , Péptidos Cíclicos/farmacología , Análisis de Supervivencia , Resultado del TratamientoRESUMEN
BACKGROUND: Insulin-dependent Type 1 diabetes (T1D) is a devastating autoimmune disease that destroys beta cells within the pancreatic islets and afflicts over 10 million people worldwide. These patients face life-long risks for blindness, cardiovascular and renal diseases, and complications of insulin treatment. New therapies that protect islets from autoimmune destruction and allow continuing insulin production are needed. Increasing evidence regarding the pathomechanism of T1D indicates that islets are destroyed by the relentless attack by autoreactive immune cells evolving from an aberrant action of the innate, in addition to adaptive, immune system that produces islet-toxic cytokines, chemokines, and other effectors of islet inflammation. We tested the hypothesis that targeting nuclear import of stress-responsive transcription factors evoked by agonist-stimulated innate and adaptive immunity receptors would protect islets from autoimmune destruction. PRINCIPAL FINDINGS: Here we show that a first-in-class inhibitor of nuclear import, cSN50 peptide, affords in vivo islet protection following a 2-day course of intense treatment in NOD mice, which resulted in a diabetes-free state for one year without apparent toxicity. This nuclear import inhibitor precipitously reduces the accumulation of islet-destructive autoreactive lymphocytes while enhancing activation-induced cell death of T and B lymphocytes derived from autoimmune diabetes-prone, non-obese diabetic (NOD) mice that develop T1D. Moreover, in this widely used model of human T1D we noted attenuation of pro-inflammatory cytokine and chemokine production in immune cells. CONCLUSIONS: These results indicate that a novel form of immunotherapy that targets nuclear import can arrest inflammation-driven destruction of insulin-producing beta cells at the site of autoimmune attack within pancreatic islets during the progression of T1D.
Asunto(s)
Núcleo Celular/efectos de los fármacos , Diabetes Mellitus Tipo 1/metabolismo , Modelos Animales de Enfermedad , Islotes Pancreáticos/efectos de los fármacos , Animales , Núcleo Celular/metabolismo , Diabetes Mellitus Tipo 1/inmunología , Inmunidad Innata , Interleucina-11/metabolismo , Interleucina-5/metabolismo , Islotes Pancreáticos/inmunología , Islotes Pancreáticos/metabolismo , Ratones , Ratones Endogámicos NOD , Transducción de SeñalRESUMEN
In this study we performed comparisons of pulmonary responses between two different respiratory syncytial virus (RSV) antigenic subgroup A strains, A2 and Line 19. Line 19 strain induced significant dose-responsive airway hyperreactivity (AHR) in BALB/c mice at days 6 and 9 after infection, whereas the A2 strain induced no AHR at any dose. Histological examination indicated that A2 induced no goblet cell hyper/metaplasia, whereas the Line 19 induced goblet cell expansion and significant increases in gob5 and MUC5AC mRNA and protein levels in vivo. When examining cytokine responses, A2 strain induced significant interleukin (IL)-10 expression, whereas Line 19 strain induced significant IL-13 expression. When IL-13-/- mice were infected with Line 19 RSV, the AHR responses were abrogated along with gob5 gene expression. There was little difference in viral titer throughout the infection between the line 19- and A2-infected mice. However, the A2 strain grew to significantly higher titers than the Line 19 strain in HEp-2 cells in vitro. Thus, RSV Line 19-induced airway dysfunction does not correlate with viral load in vivo. These data demonstrate that different RSV strains of the same antigenic subgroup can elicit differential immune responses that impact the phenotypic expression of RSV-induced illness.
Asunto(s)
Hiperreactividad Bronquial/inmunología , Infecciones por Virus Sincitial Respiratorio/inmunología , Virus Sincitiales Respiratorios/inmunología , Animales , Hiperreactividad Bronquial/patología , Hiperreactividad Bronquial/fisiopatología , Hiperreactividad Bronquial/virología , Línea Celular , Relación Dosis-Respuesta Inmunológica , Regulación de la Expresión Génica/inmunología , Células Caliciformes/inmunología , Células Caliciformes/patología , Células Caliciformes/virología , Hiperplasia/inmunología , Hiperplasia/patología , Hiperplasia/fisiopatología , Hiperplasia/virología , Interleucina-13/deficiencia , Interleucina-13/inmunología , Ratones , Ratones Noqueados , Mucina 5AC , Mucinas/inmunología , Infecciones por Virus Sincitial Respiratorio/patología , Infecciones por Virus Sincitial Respiratorio/fisiopatología , Especificidad de la EspecieRESUMEN
Nonselective cyclooxygenase (COX) inhibition during the development of allergic disease in a murine model causes an increase in type 2 cytokines and lung eosinophilia; however, the mechanisms responsible for this augmented allergen-induced inflammation have not been examined. Ab depletion of CD4 and CD8 cells revealed that the heightened allergic inflammation caused by COX inhibition was CD4, but not CD8, dependent. Allergen sensitization and airway challenge alone led to undetectable levels of IL-5 and IL-13 in the lungs of IL-4, IL-4Ralpha, and STAT6 knockout (KO) mice, but COX inhibition during the development of allergic inflammation resulted in wild-type levels of IL-5 and IL-13 and heightened airway eosinophilia in each of the three KO mice. These results indicate that the effect of COX inhibition was independent of signaling through IL-4, IL-4Ralpha, and STAT6. However, whereas COX inhibition increased IgE levels in allergic wild-type mice, IgE levels were undetectable in IL-4, IL-4Ralpha, and STAT6 KO mice, suggesting that IL-13 alone is not a switch factor for IgE synthesis in this model. These results illustrate the central role played by products derived from the COX pathway in the regulation of allergic immune responses.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Hipersensibilidad/inmunología , Prostaglandina-Endoperóxido Sintasas/inmunología , Transactivadores/inmunología , Animales , Inhibidores de la Ciclooxigenasa/farmacología , Citocinas/análisis , Femenino , Citometría de Flujo , Inmunoglobulina E/biosíntesis , Indometacina/farmacología , Inflamación/inmunología , Interleucina-13/análisis , Interleucina-13/inmunología , Interleucina-5/análisis , Interleucina-5/inmunología , Enfermedades Pulmonares/inmunología , Ratones , Ratones Noqueados , Ovalbúmina/inmunología , Prostaglandina-Endoperóxido Sintasas/metabolismo , Receptores de Interleucina-4/genética , Receptores de Interleucina-4/inmunología , Factor de Transcripción STAT6 , Transactivadores/genéticaRESUMEN
BACKGROUND: Respiratory syncytial virus (RSV) is the leading infectious cause of respiratory failure and wheezing in infants and young children. Prematurity is the greatest risk factor for severe RSV-induced disease, and recent studies suggest that premature children have lower levels of the type I IFNs (alpha/beta), for which signal transducer and activator of transcription (STAT) 1 is a critical intracellular signaling molecule. OBJECTIVE: We hypothesized that RSV infection in STAT 1 knockout (STAT 1 KO) mice would result in both increased airway resistance and airway hyperresponsiveness. METHODS: Wild-type (WT) and STAT 1 KO mice on a BALB/c background were either RSV or mock infected. Phenotypic response to infection was assessed by means of plethysmography, immunohistochemistry, and lung cytokine measurement. RESULTS: We found that STAT 1 KO mice infected with RSV (STAT 1 KO-RSV) had greater baseline lung resistance (P=.05) and airway responsiveness (P<.001) than mock-infected STAT 1 KO (STAT 1 KO-MOCK), RSV-infected wild type (WT-RSV), and mock-infected wild type (WT-MOCK) mice. In addition, the STAT 1 KO-RSV mice showed induction of mucus production and expression of gob-5 and Muc5ac, conditions not present in any of the other 3 groups. IL-17, a cytokine that regulates Muc5ac expression, was expressed in the lungs of the STAT 1 KO-RSV mice, whereas lung levels of IL-17 were undetectable in the remaining groups. Expression of the IL-23-specific p19 subunit was also increased in the STAT 1 KO-RSV mice but not in the WT-RSV mice. CONCLUSION: These results show that STAT 1 has an important regulatory role in RSV-induced alteration of airway function.
Asunto(s)
Proteínas de Unión al ADN/deficiencia , Interleucina-17/biosíntesis , Pulmón/virología , Moco/metabolismo , Infecciones por Virus Sincitial Respiratorio/fisiopatología , Transactivadores/deficiencia , Animales , Western Blotting , Hiperreactividad Bronquial/etiología , Líquido del Lavado Bronquioalveolar/citología , Canales de Cloruro/biosíntesis , Eosinofilia/etiología , Femenino , Inmunohistoquímica , Pulmón/fisiopatología , Masculino , Ratones , Ratones Noqueados , Mucina 5AC , Mucinas/biosíntesis , Mucoproteínas/biosíntesis , Pruebas de Función Respiratoria , Virus Sincitial Respiratorio Humano , Factor de Transcripción STAT1RESUMEN
Recent reports have linked mutations in the surfactant protein C gene (SFTPC) to familial forms of pulmonary fibrosis, but it is uncertain whether deficiency of mature SP-C contributes to disease pathogenesis. In this study, we evaluated bleomycin-induced lung fibrosis in mice with genetic deletion of SFTPC. Compared with wild-type (SFTPC+/+) controls, mice lacking surfactant protein C (SFTPC-/-) had greater lung neutrophil influx at 1 week after intratracheal bleomycin, greater weight loss during the first 2 weeks, and increased mortality. At 3 and 6 weeks after bleomycin, lungs from SFTPC-/- mice had increased fibroblast numbers, augmented collagen accumulation, and greater parenchymal distortion. Furthermore, resolution of fibrosis was delayed. Although remodeling was near complete in SFTPC+/+ mice by 6 weeks, SFTPC-/- mice did not return to baseline until 9 weeks after bleomycin. By terminal dUTP nick-end labeling staining, widespread cell injury was observed in SFTPC-/- and SFTPC+/+ mice 1 week after bleomycin; however, ongoing apoptosis of epithelial and interstitial cells occurred in lungs of SFTPC-/- mice, but not SFTPC+/+ mice, 6 weeks after bleomycin. Thus, SP-C functions to limit lung inflammation, inhibit collagen accumulation, and restore normal lung structure after bleomycin.
Asunto(s)
Fibrosis Pulmonar/patología , Proteína C Asociada a Surfactante Pulmonar/fisiología , Animales , Apoptosis , Bleomicina/toxicidad , Células/patología , Colágeno/análisis , Modelos Animales de Enfermedad , Fibroblastos , Hidroxiprolina/análisis , Etiquetado Corte-Fin in Situ , Recuento de Leucocitos , Pulmón/patología , Ratones , Ratones Noqueados , Neutrófilos , Peroxidasa/análisis , Fibrosis Pulmonar/inducido químicamente , Fibrosis Pulmonar/genética , Proteína C Asociada a Surfactante Pulmonar/genética , Pérdida de PesoRESUMEN
We performed polymerase chain reaction analysis, for Mycobacterium species 16S rRNA, rpoB, and IS6110 sequences, on 25 tissue specimens from patients with sarcoidosis and on 25 control tissue specimens consisting of mediastinal or cervical lymph nodes and lung biopsies. Mycobacterium species 16S rRNA sequences were amplified from 12 (48%) rpoB sequences and from 6 (24%) of the sarcoidosis specimens. In total, 16S rRNA or rpoB sequences were amplified from 15 sarcoidosis specimens (60%) but were not detected in any of the control tissues (p=0.00002, chi square). In three specimens, the sequences resembled Mycobacterium species other than M. tuberculosis. All specimens with sequences consistent with M. tuberculosis were negative for IS6110. We provide evidence that one of a variety of Mycobacterium species, especially organisms resembling M. tuberculosis, is found in most patients with sarcoidosis.
Asunto(s)
ADN Bacteriano/análisis , Mycobacterium/genética , Mycobacterium/aislamiento & purificación , Sarcoidosis/microbiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , ADN Bacteriano/genética , Femenino , Genes Bacterianos/genética , Humanos , Ganglios Linfáticos/microbiología , Masculino , Persona de Mediana Edad , Mycobacterium/clasificación , Mycobacterium tuberculosis/clasificación , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/aislamiento & purificación , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/análisis , ARN Ribosómico 16S/genética , Sensibilidad y Especificidad , Especificidad de la EspecieRESUMEN
Stimulation of macrophages with lipopolysaccharide (LPS) leads to the production of cytokines that elicit massive liver apoptosis. We investigated the in vivo role of stress-responsive transcription factors (SRTFs) in this process focusing on the precipitating events that are sensitive to a cell-permeant peptide inhibitor of SRTF nuclear import (cSN50). In the absence of cSN50, mice challenged with LPS displayed very early bursts of inflammatory cytokines/chemokines, tumor necrosis factor alpha (1 h), interleukin 6 (2 h), interleukin 1 beta (2 h), and monocyte chemoattractant protein 1 (2 h). Activation of both initiator caspases 8 and 9 and effector caspase 3 was noted 4 h later when full-blown DNA fragmentation and chromatin condensation were first observed (6 h). At this time an increase of pro-apoptotic Bax gene expression was observed. It was preceded by a decrease of anti-apoptotic Bcl2 and BclX(L) gene transcripts. Massive apoptosis was accompanied by microvascular injury manifested by hemorrhagic necrosis and a precipitous drop in blood platelets observed at 6 h. An increase in fibrinogen/fibrin degradation products and a rise in plasminogen activator inhibitor 1 occurred between 4 and 6 h. Inhibition of SRTFs nuclear import with the cSN50 peptide abrogated all these changes and increased survival from 7 to 71%. Thus, the nuclear import of SRTFs induced by LPS is a prerequisite for activation of the genetic program that governs cytokines/chemokines production, liver apoptosis, microvascular injury, and death. These results should facilitate the rational design of drugs that protect the liver from inflammation-driven apoptosis.
Asunto(s)
Transporte Activo de Núcleo Celular/fisiología , Apoptosis/efectos de los fármacos , Lipopolisacáridos/farmacología , Hígado , Péptidos/farmacología , Factores de Transcripción/metabolismo , Animales , Apoptosis/fisiología , Caspasas/metabolismo , Línea Celular , Quimiocinas/inmunología , Citocinas/inmunología , Femenino , Hepatocitos/metabolismo , Lipopolisacáridos/administración & dosificación , Lipopolisacáridos/inmunología , Hígado/efectos de los fármacos , Hígado/enzimología , Hígado/inmunología , Hígado/patología , Macrófagos/citología , Macrófagos/inmunología , Ratones , Ratones Endogámicos C57BL , Péptidos/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Tasa de Supervivencia , Proteína X Asociada a bcl-2 , Proteína bcl-XRESUMEN
Nonselective cyclooxygenase (COX) inhibition during allergic sensitization with ovalbumin in a murine model leads to an increase in the Type 2 cytokines interleukin-5 (IL-5) and IL-13; however, the effect of selective COX-1 and COX-2 inhibitors on these cytokines is unknown. We found that COX-1 protein was constitutively expressed in lung tissue. Expression of COX-1 protein did not increase with ovalbumin sensitization, but expression of COX-2 protein did. Ovalbumin-sensitized mice treated with either selective COX-1 inhibitor SC58560 (OVA-COX-1 inhibitor) or selective COX-2 inhibitor SC58236 (OVA-COX-2 inhibitor) had significantly greater airway hyperresponsiveness (p < 0.05) and higher levels of IL-13 (p < 0.05) in lung supernatants than did untreated mice that were ovalbumin sensitized (OVA). Lung mRNA levels for the chemokine receptors CCR1 through CCR5 (expressed on eosinophils, basophils, lymphocytes, and dendritic cells) were increased in the OVA-COX-2 inhibitor and OVA-indomethacin groups. We conclude that in the BALB/c mouse, COX inhibition with either a COX-1 or COX-2 inhibitor during allergen sensitization augments production of IL-13 and increases airway hyperresponsiveness.
Asunto(s)
Hiperreactividad Bronquial/fisiopatología , Inhibidores de la Ciclooxigenasa/farmacología , Isoenzimas/antagonistas & inhibidores , Compuestos Orgánicos , Pirazoles , Sulfonamidas , Animales , Ácido Araquidónico/metabolismo , Hiperreactividad Bronquial/inmunología , Hiperreactividad Bronquial/metabolismo , Pruebas de Provocación Bronquial , Líquido del Lavado Bronquioalveolar/química , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa 2 , Dinoprostona/análisis , Femenino , Inmunización , Interleucinas/metabolismo , Isoenzimas/metabolismo , Pulmón/metabolismo , Proteínas de la Membrana , Cloruro de Metacolina , Ratones , Ratones Endogámicos BALB C , Ovalbúmina/inmunología , Prostaglandina-Endoperóxido Sintasas/metabolismo , Receptores de Quimiocina/metabolismoRESUMEN
Respiratory syncytial virus (RSV) is associated with wheezing and childhood asthma. We previously reported that RSV infection prolongs methacholine-induced airway hyperresponsiveness in ovalbumin (OVA)-sensitized mice. In addition, allergically sensitized RSV-infected (OVA/RSV) mice had more abundant airway epithelial mucus production compared with OVA mice 14 days after infection, whereas there was almost no mucus in mice that were only RSV infected. We hypothesized that this increased mucus was associated with mucosal expression of Muc5ac, a mucus gene expression in airways, and gob-5, a member of the Ca(2)(+)-activated chloride channel family. By histochemical analysis, we found that there was significantly increased staining for gob-5 and Muc5ac in the airways of OVA/RSV mice compared with either OVA mice or allergically sensitized mice that were challenged with inactivated RSV, and virtually no detectable staining in the RSV group. These findings were confirmed by Western blot analysis. The increased mucus expression in the OVA/RSV group was associated with increased lung levels of interleukin-17, a factor known to stimulate airway mucin gene expression. The impact of virus infection combined with allergic inflammation on mucus production may partially explain the more severe disease and airway hyperresponsiveness associated with RSV in the setting of atopy.