RESUMEN
BACKGROUND: The clinical effectiveness of ultraviolet light (UV) disinfection remains unclear. This study aimed to investigate the effect of adding pulsed xenon UV (PX-UV) disinfection to the terminal cleaning protocol on the rate of methicillin-resistant Staphylococcus aureus (MRSA) acquisition at a Japanese hospital. METHODS: The use of a PX-UV disinfection device was added to the manual terminal cleaning protocol applied after the discharge or transfer of patients treated in the intensive and high care units. We used a Poisson regression model to examine the incidence of MRSA acquisition, based on the study period, PX-UV intervention status, unit type, and the rate of consumption of alcohol-based hand rub (ABHR). RESULTS: Approximately 86% of the rooms in the intervention units were terminally disinfected with the PX-UV device. In the intervention units, the incidence of MRSA acquisition decreased from 3.56 per 1,000 patient-days in the nonintervention period to 2.21 per 1,000 patient-days in the intervention period. Moreover, the use of PX-UV disinfection decreased the risk of MRSA acquisition (incident rate ratio: 0.556; 95% confidence interval, 0.309-0.999; Pâ¯=â¯.0497). ABHR consumption did not affect the risk of MRSA acquisition. CONCLUSIONS: Adding PX-UV disinfection to terminal manual cleaning reduced the rate of MRSA acquisition.
Asunto(s)
Infección Hospitalaria , Staphylococcus aureus Resistente a Meticilina , Infección Hospitalaria/prevención & control , Desinfección , Hospitales , Humanos , Rayos Ultravioleta , XenónRESUMEN
BACKGROUND: The hospital environment is an important source of multidrug-resistant organisms such as methicillin-resistant Staphylococcus aureus (MRSA). Here, we evaluated the efficacy of pulsed xenon ultraviolet (PX-UV) disinfection in addition to manual cleaning in a Japanese hospital. METHODS: Environmental samples were collected from inpatient rooms that had been occupied for at least 48 hours by patients infected or colonized with MRSA. High-touch surfaces from 11 rooms were sampled before and after manual cleaning and then after PX-UV disinfection. Changes in bacterial counts and in the number of aerobic bacteria (AB)- and MRSA-positive samples between sampling points were assessed. The time taken to complete PX-UV treatment of patient rooms was also recorded. RESULTS: A total of 306 samples were collected. PX-UV disinfection resulted in a significant decrease in abundance of AB and MRSA (mean colony-forming units 14.4 ± 38.7 to 1.7 ± 6.1, P < .001 and 1.1 ± 3.9 to 0.3 ± 2.0, P < .001, respectively) and in the number of AB- and MRSA-positive samples (58.8%-28.4%, Pâ¯=â¯.001 and 19.6%-3.9%, P < .001, respectively) compared with manual cleaning. The median time of in-room use of the PX-UV device was 20 minutes. CONCLUSIONS: The addition of PX-UV disinfection to the manual cleaning process significantly reduced AB and MRSA contamination of high-touch surfaces in hospital inpatient rooms.
Asunto(s)
Desinfección/métodos , Fómites/microbiología , Staphylococcus aureus Resistente a Meticilina/efectos de la radiación , Rayos Ultravioleta , Xenón , Carga Bacteriana , Hospitales , Humanos , Japón , Habitaciones de PacientesRESUMEN
CD11c, a member of the ß(2) integrin family of adhesion molecule, is expressed on the surface of myeloid lineages and activated lymphoid cells and forms a heterodimeric receptor with CD18. We analyzed the mouse CD11c promoter structure to elucidate the transcriptional regulation in dendritic cells (DCs). By reporter assay, the -84/-65 region was identified to be essential for activity of the mouse CD11c promoter in the mouse bone marrow-derived (BM) DCs and monocyte cell line RAW264.7. An electrophoretic mobility shift assay using a number of antibodies against transcription factors revealed that the target region was recognized by a complex including JunD and Fra2, which are transcription factors belonging to the AP-1 family. The direct interaction of JunD and Fra2 with the CD11c promoter was further confirmed by a chromatin immunoprecipitation assay using CD11c-positive cells purified from BMDCs. Finally, mouse JunD and/or Fra2 siRNA was introduced into BMDCs to evaluate the involvement of these factors against CD11c transcription and found that Fra2 siRNA reduced cell surface expression level of CD11c. These results indicate that AP-1 composed with JunD and Fra2 protein plays a primary role in enhancing the transcription level of the CD11c gene in DC.
Asunto(s)
Antígeno CD11c/genética , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Antígeno 2 Relacionado con Fos/metabolismo , Proteínas Proto-Oncogénicas c-jun/metabolismo , Factor de Transcripción AP-1/metabolismo , Animales , Secuencia de Bases , Sitios de Unión/genética , Línea Celular , Mapeo Cromosómico , Cartilla de ADN/genética , Elementos de Facilitación Genéticos , Antígeno 2 Relacionado con Fos/antagonistas & inhibidores , Antígeno 2 Relacionado con Fos/química , Antígeno 2 Relacionado con Fos/genética , Regulación de la Expresión Génica , Ratones , Datos de Secuencia Molecular , Complejos Multiproteicos/química , Complejos Multiproteicos/metabolismo , Regiones Promotoras Genéticas , Proteínas Proto-Oncogénicas c-jun/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-jun/química , Proteínas Proto-Oncogénicas c-jun/genética , ARN Interferente Pequeño/genética , Factor de Transcripción AP-1/químicaRESUMEN
BACKGROUND: IgE-mediated immediate-type skin reaction shows a diurnal rhythm, although the precise mechanisms remain uncertain. Period2 (Per2) is a key circadian gene that is essential for endogenous clockworks in mammals. OBJECTIVE: This study investigated whether Per2 regulates a time-of-day-dependent variation in IgE-mediated immediate-type skin reaction. METHODS: The kinetics of a passive cutaneous anaphylactic reaction were compared between wild-type mice and mice with a loss-of-function mutation of Per2 (mPer2(m/m) mice). The effects of adrenalectomy, aging, and dexamethasone on the kinetics of a passive cutaneous anaphylactic reaction were also examined. In addition, the extent of IgE-mediated degranulation in bone marrow-derived mast cells (BMMCs) was compared between wild-type and mPer2(m/m) mice. RESULTS: A time-of-day-dependent variation in a passive cutaneous anaphylactic reaction observed in wild-type mice was absent in mPer2(m/m) mice and in adrenalectomized and aged mice associated with the loss of rhythmic secretion of corticosterone. In addition, mPer2(m/m) mice showed decreased sensitivity to the inhibitory effects of dexamethasone on the passive cutaneous anaphylactic reactions. IgE-mediated degranulation in BMMCs was comparable between wild-type and mPer2(m/m) mice, but Per2 mutation decreased sensitivity to the inhibitory effects of dexamethasone on IgE-mediated degranulation in BMMCs. CONCLUSION: A circadian oscillator, Per2, regulates a time-of-day-dependent variation in a passive cutaneous anaphylactic reaction in mice. Per2 may do so by controlling the rhythmic secretion of glucocorticoid from adrenal glands and/or by gating the glucocorticoid responses of mast cells to certain times of the day (possibly when Per2 levels are high in mast cells).
Asunto(s)
Anafilaxia/genética , Ritmo Circadiano/genética , Ritmo Circadiano/inmunología , Proteínas Circadianas Period/genética , Piel/inmunología , Anafilaxia/inmunología , Animales , Separación Celular , Corticosterona/sangre , Citometría de Flujo , Inmunoglobulina E/sangre , Ratones , Ratones Endogámicos ICR , Ratones Mutantes , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Increasing evidence suggests that the aryl hydrocarbon receptor (AhR) pathway has an important role in the regulation of inflammatory responses. Most recently, we have shown that the activation of the AhR pathway by a potent AhR agonist inhibits the development of dextran sodium sulfate (DSS)-induced colitis, a model of human ulcerative colitis, by the induction of prostaglandin E2 (PGE2) in the large intestine. Because several strains of probiotic lactic acid bacteria have been reported to inhibit DSS-induced colitis by unidentified mechanisms, we hypothesized that particular strains of lactic acid bacterium might have the potential to activate the AhR pathway, thereby inhibiting DSS-induced colitis. This study investigated whether there are specific lactic acid bacterial strains that can activate the AhR pathway, and if so, whether this AhR-activating potential is associated with suppression of DSS-induced colitis. By using AhR signaling reporter cells, we found that Lactobacillus bulgaricus OLL1181 had the potential to activate the AhR pathway. OLL1181 also induced the mRNA expression of cytochrome P450 family 1A1 (CYP1A1), a target gene of the AhR pathway, in human colon cells, which was inhibited by the addition of an AhR antagonist, α-naphthoflavon (αNF). In addition, mice treated orally with OLL1181 showed an increase in CYP1A1 mRNA expression in the large intestine and amelioration of DSS-induced colitis. Thus, OLL1181 can induce activation of the intestinal AhR pathway and inhibit DSS-induced colitis in mice. This strain of lactic acid bacterium has therefore the potential to activate the AhR pathway, which may be able to suppress colitis.
Asunto(s)
Colitis Ulcerosa/prevención & control , Citocromo P-450 CYP1A1/metabolismo , Lactobacillus/fisiología , Probióticos , Receptores de Hidrocarburo de Aril/metabolismo , Animales , Benzoflavonas/farmacología , Línea Celular Tumoral , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/metabolismo , Colon/metabolismo , Colon/microbiología , Citocromo P-450 CYP1A1/genética , Sulfato de Dextran/farmacología , Femenino , Humanos , Ratones , Ratones Endogámicos C57BL , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de SeñalRESUMEN
To evaluate the effects of the transcription factor GATA-1 on determining cell fate between dendritic cell (DC) and mast cell (MC) lineages, GATA-1 was exogenously expressed in bone marrow-derived (BM) DCs. Exogenous expression of GATA-1 by a retrovirus in BMDCs inhibited expression of CD11c, CD80, CD86, and major histocompatibility complex class II with suppression of antigen-presenting ability and morphological changes toward granulocyte-like cells. Transcription of MC proteases and c-kit was markedly upregulated by GATA-1. Expression of IRF-4 and -8 was markedly suppressed, whereas PU.1 mRNA level was not affected by GATA-1. Chromatin immunoprecipitation assay showed that recruitment of PU.1 on the IRF-8 promoter was reduced in GATA-1-expressing DCs. These results indicate that GATA-1 suppresses PU.1 function but not PU.1 transcription. Thus, GATA-1 appears to determine cell fate by regulating several cell-specific transcription factors.
Asunto(s)
Células de la Médula Ósea/metabolismo , Células Dendríticas/metabolismo , Factor de Transcripción GATA1/genética , Regulación de la Expresión Génica/fisiología , Animales , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Western Blotting , Diferenciación Celular , Linaje de la Célula , Células Cultivadas , Inmunoprecipitación de Cromatina , Citometría de Flujo , Factores Reguladores del Interferón/genética , Factores Reguladores del Interferón/metabolismo , Mastocitos/metabolismo , Ratones , Ratones Endogámicos BALB C , Regiones Promotoras Genéticas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Mast cells (MCs) are activated upon stimulation via TLRs or FcepsilonRI, contributing to immune protection and/or leading to allergic diseases. In the present study, the effects of Trichostatin A (TSA) on the activation of MCs were analyzed with bone marrow-derived (BM) MCs. TSA increased the transcription and protein secretion of IL-6 in case of LPS-stimulation, in contrast to the suppressive effect on IgE-mediated activation of BMMCs. Chromatin immunoprecipitation assay showed IgE-mediated signaling-specific suppression of transcription factors recruitment to the IL-6 promoter. TSA-treatment inhibited nuclear translocation of NF-kappaB following IgE-mediated, but not LPS-induced activation in MCs.
Asunto(s)
Ácidos Hidroxámicos/farmacología , Interleucina-6/genética , Interleucina-6/metabolismo , Mastocitos/metabolismo , FN-kappa B/metabolismo , Lipopolisacáridos/farmacología , Mastocitos/efectos de los fármacos , FN-kappa B/genética , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The aryl hydrocarbon receptor (AhR) recognizes numerous small xenobiotic and natural molecules, such as dioxin and natural chemicals, and is involved in the metabolism of these compounds. AhR also has a regulatory role in inflammatory responses. This study investigated whether the activation of the AhR pathway affects dextran sodium sulfate (DSS)-induced colitis, an ulcerative colitis-like model, in mice. DSS-induced colitis was ameliorated by pretreatment with a potent AhR activator, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), in mice. In addition, the mice pretreated with TCDD showed increased prostaglandin E2 (PGE2) production in the colon, and inhibition of PGE2 production by indomethacin abrogated the inhibitory effects of TCDD on DSS-induced colitis. Collectively, the activation of the AhR pathway by TCDD may ameliorate DSS-induced colitis, at least in part, through PGE2 production.
Asunto(s)
Colitis/inmunología , Colon/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Dinoprostona/metabolismo , Dibenzodioxinas Policloradas/administración & dosificación , Receptores de Hidrocarburo de Aril/genética , Animales , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/fisiopatología , Colitis Ulcerosa/inmunología , Colon/efectos de los fármacos , Colon/inmunología , Colon/patología , Citocromo P-450 CYP1A1/genética , Sulfato de Dextran/administración & dosificación , Dinoprostona/genética , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Humanos , Indometacina/administración & dosificación , Ratones , Ratones Endogámicos C57BL , Receptores de Hidrocarburo de Aril/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/inmunologíaRESUMEN
Both thymic stromal lymphopoietin (TSLP) and IL-13 are essential cytokines for the development of allergic inflammation. However, a causal link between TSLP and IL-13 has not yet been fully elucidated. This study aimed to investigate whether IL-13 induces TSLP expression and whether the induction contributes to the development of allergic inflammation. We found that IL-13 induced TSLP expression in mouse nasal tissue specimens in a Stat6-dependent manner. In addition, intranasal challenge of mice with IL-13 induced TSLP expression in the nasal epithelium. Importantly, intranasal IL-13 challenge induced eosinophilia and goblet cell hyperplasia in the nasal mucosa in mice, which was inhibited by the blockade of TSLP activity with anti-TSLP Ab. These findings suggest that TSLP is an important mediator of IL-13-driven allergic inflammation in the nasal mucosa. Taken together with the recent findings that IL-13 is a critical downstream element for TSLP-driven allergic inflammation, TSLP may function both upstream and downstream of IL-13, thus providing an additional rationale as to why TSLP plays such a central role in the development of allergic inflammation.
Asunto(s)
Citocinas/inmunología , Hipersensibilidad/inmunología , Inflamación/inmunología , Mucosa Nasal/inmunología , Animales , Ensayo de Inmunoadsorción Enzimática , Femenino , Interleucina-13/inmunología , Ratones , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfopoyetina del Estroma TímicoRESUMEN
Transforming growth factor-beta (TGFbeta) is abundant in mammalian milk in a latent form. However, whether the latent form of TGFbeta in human milk is converted to the active form in vivo remains uncertain. To address this issue, we first investigated whether latent TGFbeta or human milk-borne latent TGFbeta was activated in an in vitro assay, simulating the effects of gastric acid. We then tested whether gastric acid was necessary for the activation of orally administered latent TGFbeta or human milk-borne latent TGFbeta in mice by inhibiting gastric acidity with cimetidine, an antagonist of H2-receptors. Latent TGFbeta or human milk-borne latent TGFbeta increased Smad-responsive promoter activity in MFB-F11 reporter cells at pH 1.2, but not at pH 7.0, regardless of the presence or absence of the gastric protease pepsin. In mice treated orally with latent TGFbeta (5 microg/mouse), the phosphorylation of Smad2 and TGFbeta target gene mRNA expression (TGFbeta and Smad7) was increased in the small intestine (P < 0.05) and this effect was inhibited by cimetidine (100 mg/kg, intraperitoneally). Similarly, mice treated orally with 1200 microL/d of human milk containing latent TGFbeta (3 microg/L) for 2 wk had increased TGFbeta and Smad7 mRNA expression in the small intestine (P < 0.05) and this was inhibited by the antiacid treatment. Therefore, the latent form of TGFbeta, such as TGFbeta in human milk, can be activated by gastric acid following oral administration in mice. This process may be involved in the conversion of human milk-borne latent TGFbeta to the active form in vivo.
Asunto(s)
Ácido Gástrico/metabolismo , Leche Humana/metabolismo , Proteína Smad2/metabolismo , Proteína smad7/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Administración Oral , Adulto , Animales , Antiulcerosos/farmacología , Cimetidina/farmacología , Femenino , Genes Reporteros , Humanos , Concentración de Iones de Hidrógeno , Intestino Delgado/efectos de los fármacos , Intestino Delgado/metabolismo , Ratones , Ratones Endogámicos BALB C , Fosforilación/fisiología , ARN Mensajero/metabolismo , Transducción de Señal/efectos de los fármacos , Proteína Smad2/genética , Proteína smad7/genética , Factor de Crecimiento Transformador beta/genética , Adulto JovenRESUMEN
Subtilase cytotoxin (SubAB) is the prototype of a newly identified family of AB(5) cytotoxins produced by Shiga toxigenic Escherichia coli. SubAB specifically cleaves the essential endoplasmic reticulum (ER) chaperone BiP (GRP78), resulting in the activation of ER stress-induced unfolded protein response (UPR). We have recently shown that the UPR following ER stress can suppress cellular responses to inflammatory stimuli during the later phase, in association with inhibition of NF-kappaB activation. These findings prompted us to hypothesize that SubAB, as a selective UPR inducer, might have beneficial effects on inflammation-associated pathology via a UPR-dependent inhibition of NF-kappaB activation. The pretreatment of a mouse macrophage cell line, RAW264.7, with a subcytotoxic dose of SubAB-triggered UPR and inhibited LPS-induced MCP-1 and TNF-alpha production associated with inhibition of NF-kappaB activation. SubA(A272)B, a SubAB active site mutant that cannot induce UPR, did not show such effects. In addition, pretreatment with a sublethal dose of SubAB, but not SubA(A272)B, protected the mice from LPS-induced endotoxic lethality associated with reduced serum MCP-1 and TNF-alpha levels and also prevented the development of experimental arthritis induced by LPS in mice. Collectively, although SubAB has been identified originally as a toxin associated with the pathogenesis of hemolytic uremic syndrome, the unique ability of SubAB to selectively induce the UPR may have the potential to prevent LPS-associated inflammatory pathology under subcytotoxic conditions.
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Retículo Endoplásmico/patología , Proteínas de Escherichia coli/farmacología , Inflamación/prevención & control , Chaperonas Moleculares/efectos de los fármacos , Pliegue de Proteína/efectos de los fármacos , Subtilisinas/farmacología , Animales , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/prevención & control , Línea Celular , Citotoxinas , Chaperón BiP del Retículo Endoplásmico , Proteínas de Escherichia coli/administración & dosificación , Inflamación/inducido químicamente , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Chaperonas Moleculares/metabolismo , FN-kappa B/antagonistas & inhibidores , Subtilisinas/administración & dosificaciónRESUMEN
A major house dust mite allergen Der f 1 belongs to the papain-like cysteine protease family. This study investigated whether Der f 1 can cleave the latency-associated peptide (LAP) of transforming growth factor (TGF)-beta via its proteolytic activity and activate latent TGF-beta. We found that Der f 1 can cleave LAP and induce the activation of latent TGF-beta, leading to functional Smad signaling. Importantly, these actions of Der f 1 were inhibited by cysteine protease inhibitor E64 or inactivation of the protease activity by heat. Thus, latent TGF-beta may be a direct target of Der f 1 protease activity.
Asunto(s)
Alérgenos/metabolismo , Antígenos Dermatofagoides/metabolismo , Cisteína Endopeptidasas/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Alérgenos/efectos de los fármacos , Animales , Antígenos Dermatofagoides/administración & dosificación , Antígenos Dermatofagoides/efectos de los fármacos , Proteínas de Artrópodos , Cisteína Endopeptidasas/inmunología , Inhibidores de Cisteína Proteinasa/farmacología , Expresión Génica , Leucina/análogos & derivados , Leucina/farmacología , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Ratones , Ratones Endogámicos BALB C , Pyroglyphidae/enzimología , Pyroglyphidae/inmunología , Transducción de Señal , Proteínas Smad/genética , Proteínas Smad/metabolismoRESUMEN
OBJECTIVE: Intraoperative auditory brainstem response (ABR)-monitoring is useful for hearing preservation in patients undergoing cerebellopontine angle surgery. Prolongation of the latency of wave V, for example, is observed under surgical stress such as cerebellar retraction. We analyzed intraoperative ABR findings to study the neurophysiological mechanism(s) underlying latency prolongation. METHODS: The ABR recorded during microvascular decompression surgery was studied in 18 patients with hemifacial spasm. We measured each trace of the ABR records, both the latency of each wave and some interpeak latencies. We also analyzed their waveforms especially in the early component, to assess changes during surgery. RESULTS: The latency of wave V varied with cerebellar retraction. The delayed latency of wave V was correlated with the prolonged interpeak latency of waves I-III. An additional wave (designated wave I') between waves I and II was appeared; it was accompanied by a prolongation in the latency of wave V. Wave I' contributed to prolongation of the interpeak latency of waves I-III, resulting in a delay in the latency of wave V. Chronological analysis revealed that the minimum latency of wave I' was the same as wave IN, suggesting that wave I' arose near the porus acusticus internus (PAI). CONCLUSION: Our study showed that cerebellar retraction may result in conduction impairment of the auditory nerve near the PAI, suggesting that the Obersteiner-Redlich zone is an electrophysiologically vulnerable site and wave I' is derived from the change in the vector of wave IN. SIGNIFICANCE: Our findings may provide neurophysiological evidence to support the theoretical model of ABR generators by Scherg and von Cramon.
Asunto(s)
Ángulo Pontocerebeloso/cirugía , Potenciales Evocados Auditivos del Tronco Encefálico/fisiología , Espasmo Hemifacial/fisiopatología , Espasmo Hemifacial/cirugía , Monitoreo Intraoperatorio , Estimulación Acústica/métodos , Adulto , Anciano , Descompresión Quirúrgica , Electroencefalografía/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Modelos Biológicos , Tiempo de Reacción/fisiología , Estudios Retrospectivos , Adulto JovenRESUMEN
Cow milk contains a large amount of an immunoregulatory cytokine, transforming growth factor-beta (TGFbeta). The present study investigated whether commercially available pasteurized cow milk retains TGFbeta activity both in vitro and in vivo. Some commercial cow milk increased TGFbeta/Smad-responsive reporter activity and induced Smad2 phosphorylation and the transcription of the TGFbeta/Smad target genes TGFbeta itself and Smad7 in vitro. Mice treated orally with 500 microL of cow milk containing TGFbeta (3 microg/L) daily for 2 wk had increased phosphorylation of Smad2 and TGFbeta and Smad7 mRNA expression in the intestine. These mice also had significantly greater serum TGFbeta concentrations than the mice treated orally with PBS. Furthermore, oral administration of 500 microL of cow milk containing TGFbeta (3 microg/L) daily for 2 wk before the induction of dextran sodium sulfate colitis and lipopolysaccharide-induced endotoxemia ameliorated tissue damage and mortality, respectively, in mice. These in vivo effects of cow milk were abrogated by the simultaneous administration of TGFbeta type I receptor kinase inhibitor with the cow milk, and they were not observed after the oral administration of cow's milk containing little TGFbeta. In humans, 1 oral challenge of 10 mL/kg cow milk containing TGFbeta (3 microg/L) increased the plasma TGFbeta concentrations at 4 h after the challenge. Thus, some commercially available pasteurized cow milk retains TGFbeta activity, which may be able to provide protection against experimental colitis and endotoxemia associated with increased intestinal and circulating TGFbeta levels.
Asunto(s)
Inflamación/prevención & control , Leche/química , Factor de Crecimiento Transformador beta/análisis , Factor de Crecimiento Transformador beta/farmacología , Adulto , Animales , Bovinos , Línea Celular Tumoral , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Sulfato de Dextran/toxicidad , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad , Factor de Crecimiento Transformador beta/sangre , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
BACKGROUND: Both active and passive smoking are considered to be risk factors for asthma development. However, the precise mechanisms involved remain elusive. Recently, thymic stromal lymphopoietin (TSLP) has been shown to play a key role in the development of T(H)2-type allergic inflammation in patients with asthma. OBJECTIVE: The aim of this study was to investigate whether there was a causal relationship between cigarette smoke exposure and TSLP expression in the lung. METHODS: We examined the effects of repeated intranasal exposure of cigarette smoke extract (CSE) on TSLP mRNA and protein expression in the mouse lung by means of real-time PCR, Western blotting, and immunohistochemistry. We also examined the effects of intranasal exposure of CSE plus ovalbumin (OVA) on T(H)2-type immune responses and lung pathology. RESULTS: Repeated exposure of CSE induced TSLP mRNA and protein expression, which was inhibited by treatment with antioxidative N-acetylcysteine and by TNF-alpha receptor I deficiency. In addition, the intranasal exposure of CSE simultaneously with OVA induced OVA-specific T(H)2-type immune responses and airway inflammation, which were inhibited by the blockade of the TSLP activity. CONCLUSION: CSE induced TSLP expression in the mouse lung in an oxidative stress-dependent and TNF-alpha receptor I-dependent manner, and when challenged simultaneously with an antigen, CSE promoted the development of airway inflammation in association with T(H)2-type immune responses.
Asunto(s)
Asma/inmunología , Mezclas Complejas/toxicidad , Citocinas/inmunología , Regulación de la Expresión Génica/inmunología , Células Th2/inmunología , Contaminación por Humo de Tabaco/efectos adversos , Acetilcisteína/farmacología , Administración Intranasal , Animales , Asma/inducido químicamente , Asma/genética , Asma/patología , Citocinas/genética , Femenino , Depuradores de Radicales Libres/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Inflamación/inducido químicamente , Inflamación/genética , Inflamación/inmunología , Inflamación/patología , Ratones , Ratones Endogámicos BALB C , Ratones Mutantes , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/genética , Estrés Oxidativo/inmunología , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Receptores Tipo I de Factores de Necrosis Tumoral/inmunología , Factores de Riesgo , Células Th2/patología , Linfopoyetina del Estroma TímicoRESUMEN
The high-affinity receptor for immunoglobulin E (IgE), FcepsilonRI, is specifically expressed in mast cells and basophils and plays a key role in IgE-mediated allergic reactions. The transcription factor Elf-1 has been previously identified to bind to the promoter of the human FcepsilonRI alpha-chain, which is essential for the function and expression of FcepsilonRI. In the present study, Elf-1 siRNA was conducted to evaluate the effects of Elf-1 on FcepsilonRI alpha-chain expression in the primary mouse mast cells, bone marrow-derived mast cells (BMMC). Introduction of Elf-1 siRNA effectively reduced expression levels of Elf-1 mRNA and protein in BMMC. Transient reporter assay showed that the knockdown of Elf-1 by siRNA resulted in increased FcepsilonRI alpha-chain promoter activity, while overexpression of Elf-1 suppressed alpha-chain promoter activity in BMMC. Elf-1 siRNA-treated BMMC exhibited marked upregulation of FcepsilonRI alpha-chain transcription, whereas beta-chain mRNA was not affected by Elf-1 siRNA. Chromatin immunoprecipitation assay showed that the amount of transcription factor PU.1, recognizing the cis-element close to the Elf-1-site on the FcepsilonRI alpha-chain promoter, was significantly increased by introduction of Elf-1 siRNA. These results indicate that Elf-1 negatively regulates FcepsilonRI alpha-chain expression by suppressing PU.1-mediated transcription of the alpha-chain in BMMC.
Asunto(s)
Efrina-A2/antagonistas & inhibidores , Regulación de la Expresión Génica , Mastocitos/metabolismo , ARN Interferente Pequeño/farmacología , Receptores de IgE/genética , Animales , Western Blotting , Médula Ósea/metabolismo , Inmunoprecipitación de Cromatina , Humanos , Luciferasas/metabolismo , Ratones , Regiones Promotoras Genéticas/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de IgE/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transactivadores/genética , Transactivadores/metabolismo , Transcripción Genética , Regulación hacia ArribaRESUMEN
The alpha-chain is a specific component of FcepsilonRI, which is essential for the cell surface expression of FcepsilonRI and the binding of IgE. Recently, two single nucleotide polymorphisms (SNPs) in the alpha-chain promoter, -315C>T and -66T>C, have been shown by statistic studies to associate with allergic diseases. The effect of -66 SNP on GATA-1-mediated promoter activity has been already indicated. In the present study, to investigate roles of the -315 SNP on the alpha-chain promoter functions, the transcription activity was evaluated by reporter assay. The alpha-chain promoter carrying -315T (minor allele) possessed significantly higher transcriptional activity than that of -315C (major allele). EMSA indicated that the transcription factor Sp1, but not Myc-associated zinc finger protein (MAZ), was bound to the -315C allele probe and that a transcription factor belonging to a high mobility group-family bound to the -315T allele probe. The chromatin immunoprecipitation assay suggested that high mobility group 1, 2, and Sp1 bound around -315 of FcepsilonRIalpha genomic DNA in vivo in the human basophil cell line KU812 with -315C/T and in human peripheral blood basophils with -315C/C, respectively. When cell surface expression level of FcepsilonRI on basophils was analyzed by flow cytometry, basophils from individuals carrying -315T allele expressed significantly higher amount of FcepsilonRI compared with those of -315C/C. The findings demonstrate that a -315 SNP significantly affects human FcepsilonRI alpha-chain promoter activity and expression level of FcepsilonRI on basophils by binding different transcription factors to the SNP site.
Asunto(s)
Proteínas del Grupo de Alta Movilidad/metabolismo , Polimorfismo de Nucleótido Simple/inmunología , Subunidades de Proteína/genética , Subunidades de Proteína/metabolismo , Receptores de IgE/genética , Receptores de IgE/metabolismo , Factor de Transcripción Sp1/metabolismo , Alelos , Animales , Basófilos/inmunología , Basófilos/metabolismo , Línea Celular Tumoral , Membrana Celular/genética , Membrana Celular/inmunología , Membrana Celular/metabolismo , Nucleótidos de Citosina/genética , Nucleótidos de Citosina/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Proteínas del Grupo de Alta Movilidad/genética , Humanos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Regiones Promotoras Genéticas/inmunología , Unión Proteica/genética , Unión Proteica/inmunología , Subunidades de Proteína/biosíntesis , Ratas , Receptores de IgE/biosíntesis , Factor de Transcripción Sp1/genética , Nucleótidos de Timina/genética , Nucleótidos de Timina/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Epithelial cell-derived thymic stromal lymphopoietin (TSLP) is a master switch for asthma or atopic dermatitis by inducing a dendritic cell-mediated Th2-type allergic inflammation. Allergic rhinitis is also pathologically characterized by Th2-type allergic inflammation. This study demonstrates that mast cells regulate the epithelial TSLP expression in allergic rhinitis. TSLP expression was found to be up-regulated predominantly in the nasal epithelium in the ovalbumin (OVA)-sensitized and -nasally challenged mouse model of allergic rhinitis, which was abolished in mast cell-deficient WBB6F1-W/W(v) in comparison with control WBB6F1-+/+ mice. Similarly, the epithelial TSLP expression was reduced in Fc receptor gamma chain (FcgammaR)-deficient mice, where the high-affinity IgE receptor (FcepsilonRI) is not expressed on mast cells, in comparison with control C57BL/6 mice. Furthermore, the administration of neutralizing TSLP antibody during the challenge phase of OVA inhibited the development of allergic rhinitis. These results suggest that the direct stimulation of epithelial cells by antigens alone may not be sufficient to induce TSLP expression in the nasal epithelium, and that mast cell regulation of epithelial TSLP expression, possibly via FcepsilonRI, plays an important role in the development of allergic rhinitis.
Asunto(s)
Citocinas/metabolismo , Mastocitos/inmunología , Mucosa Nasal/metabolismo , Hipersensibilidad Respiratoria/metabolismo , Rinitis/metabolismo , Animales , Anticuerpos/farmacología , Conducta Animal/efectos de los fármacos , Recuento de Células , Citocinas/genética , Citocinas/fisiología , Modelos Animales de Enfermedad , Células Epiteliales/metabolismo , Células Epiteliales/patología , Femenino , Células Caliciformes/patología , Mastocitos/trasplante , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mucosa Nasal/efectos de los fármacos , Mucosa Nasal/patología , Ovalbúmina/inmunología , Proteínas Proto-Oncogénicas c-kit/genética , Proteínas Proto-Oncogénicas c-kit/metabolismo , Receptores de IgG/deficiencia , Receptores de IgG/genética , Receptores de IgG/fisiología , Hipersensibilidad Respiratoria/patología , Rinitis/patología , Linfopoyetina del Estroma TímicoRESUMEN
Interleukin-12 (IL-12), a heterodimeric cytokine (p35/p40) produced mainly from macrophages and dendritic cells, is an important regulator of T-helper 1 cell responses and for host defense. We found that interferon (IFN) consensus sequence binding protein (ICSBP), which is a transcription factor essential for the expression of p40, was expressed in mouse bone marrow-derived mast cells (BMMCs). The transcription levels of p35 and p40 were increased by stimulation of BMMCs with IFN-gamma/lipopolysaccharide (LPS). IL-12 was secreted from BMMCs in response to LPS but not by FcepsilonRI cross-linking. The p40 levels in the peritoneal cavity of mast cell-deficient W/W(v) and W/W(v) reconstituted with p40(-/-) BMMCs were significantly lower than those of WBB6F(1)(+/+) and wild-type (WT) BMMC-reconstituted W/W(v) in the acute septic peritonitis model. The survival rate of W/W(v) reconstituted with p40(-/-) BMMCs was significantly decreased compared to those of WBB6F(1)(+/+) and WT-BMMC-reconstituted W/W(v), which was due to reduced production of IFN-gamma and subsequent impaired activation of neutrophils in the peritoneal cavity. Survival rate of p40(-/-) mice was also restored by adoptive transfer of WT-BMMCs. These results demonstrate that mast cells play a significant role in the production of IL-12 required for host defense. This is the first report to demonstrate that mast cells are a crucial source of functional IL-12.
Asunto(s)
Infecciones Bacterianas/metabolismo , Células Dendríticas/inmunología , Subunidad p40 de la Interleucina-12/biosíntesis , Interleucina-12/biosíntesis , Mastocitos/metabolismo , Neutrófilos/microbiología , Peritonitis/microbiología , Sepsis/sangre , Animales , Infecciones Bacterianas/patología , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Neutrófilos/metabolismoRESUMEN
The SecYEG heterotrimeric membrane protein complex functions as a channel for protein translocation across the Escherichia coli cytoplasmic membrane. SecY is the central subunit of the SecYEG complex and contains 10 transmembrane segments (TM1 to TM10). Previous mutation studies suggested that TM3 and TM4 are particularly important for SecY function. To further characterize TM3 and TM4, we introduced a series of cysteine-scanning mutations into these segments. With one exception (an unstable product), all the mutant proteins complemented the cold-sensitive growth defect of the secY39 mutant. A combination of this secY mutation and the secG deletion resulted in synthetic lethality, and the TM3 and TM4 SecY cysteine substitution mutations were examined for their ability to complement this lethality. Although they were all positive for complementation, some of the complemented cells exhibited significant retardation of protein export. The substitution-sensitive residues in TM3 can be aligned to one side of the alpha-helix, and those in TM4 revealed a tendency for residues closer to the cytosolic side of the membrane to be more severely affected. Disulfide cross-linking experiments identified a specific contact point for TM3 and SecG TM2 as well as for TM4 and SecG TM1. Thus, although TM3 and TM4 do not contain any single residue that is absolutely required, they include functionally important helix surfaces and specific contact points with SecG. These results are discussed in light of the structural information available for the SecY complex.