RESUMEN
The healing phase of contact hypersensitivity reactions is critically dependent on regulatory T cells (Tregs), but even the early inflammatory phase, that is, 6-24 hours after induction of a contact hypersensitivity reaction, is susceptible to Treg-mediated suppression. To investigate the underlying mechanisms, we injected Tregs before the challenge and analyzed the skin-infiltrating cells as early as 6 hours later. Early on, we found mainly neutrophils in the challenged skin, but only a few T cells. This influx of neutrophils was blocked by the injection of Tregs, indicating that they were able to prevent the first wave of leukocytes, which are responsible for starting an immune reaction. As an underlying mechanism, we identified that Tregs can tighten endothelial junctions by inducing intracellular cAMP, leading to protein kinase A-RhoAâdependent signaling. This eventually reorganizes endothelial junction proteins, such as Notch3, Nectin 2, Filamin B, and VE-cadherin, all of which contribute to the tightening of the endothelial barrier. In summary, Tregs prevent the leakage of proinflammatory cells from and into the tissue, which establishes a mechanism to downregulate immune reactions.
Asunto(s)
Dermatitis Alérgica por Contacto/inmunología , Endotelio Vascular/patología , Neutrófilos/inmunología , Linfocitos T Reguladores/inmunología , Animales , Comunicación Celular/inmunología , Quimiotaxis/inmunología , Dermatitis Alérgica por Contacto/patología , Modelos Animales de Enfermedad , Endotelio Vascular/inmunología , Humanos , Ratones , Cloruro de Picrilo/administración & dosificación , Cloruro de Picrilo/inmunología , Piel/irrigación sanguínea , Piel/inmunología , Piel/patologíaAsunto(s)
Dermatitis Atópica/tratamiento farmacológico , Fármacos Dermatológicos/administración & dosificación , Receptor Toll-Like 3/antagonistas & inhibidores , Administración Cutánea , Animales , Dermatitis Atópica/inmunología , Dermatitis Atópica/patología , Modelos Animales de Enfermedad , Humanos , Mediadores de Inflamación/metabolismo , Queratinocitos/efectos de los fármacos , Queratinocitos/inmunología , Queratinocitos/metabolismo , Ratones , Cloruro de Picrilo/administración & dosificación , Poli I-C/administración & dosificación , Poli I-C/inmunología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/inmunología , Piel/citología , Piel/efectos de los fármacos , Piel/inmunología , Piel/patología , Receptor Toll-Like 3/metabolismoRESUMEN
In the present study, we demonstrated that there is a direct relationship between scratching behaviors induced by itch and functional changes in the brain reward system. Using a conditional place preference test, the rewarding effect was clearly evoked by scratching under both acute and chronic itch stimuli. The induction of ΔFosB, a member of the Fos family of transcription factors, was observed in dopamine transporter (DAT)-positive dopamine neurons in the ventral tegmental area (VTA) of mice suffering from a chronic itch sensation. Based on a cellular analysis of scratching-activated neurons, these neurons highly expressed tyrosine hydroxylase (TH) and DAT genes in the VTA. Furthermore, in an in vivo microdialysis study, the levels of extracellular dopamine in the nucleus accumbens (NAcc) were significantly increased by transient scratching behaviors. To specifically suppress the mesolimbic dopaminergic pathway using pharmacogenetics, we used the TH-cre/hM4Di mice. Pharmacogenetic suppression of mesolimbic dopaminergic neurons significantly decreased scratching behaviors. Under the itch condition with scratching behaviors restricted by an Elizabethan collar, the induction of ΔFosB was found mostly in corticotropin-releasing hormone (CRH)-containing neurons of the hypothalamic paraventricular nucleus (PVN). These findings suggest that repetitive abnormal scratching behaviors under acute and chronic itch stimuli may activate mesolimbic dopamine neurons along with pleasant emotions, while the restriction of such scratching behaviors may initially induce the activation of PVN-CRH neurons associated with stress.
Asunto(s)
Prurito/fisiopatología , Prurito/psicología , Recompensa , Área Tegmental Ventral/fisiopatología , Enfermedad Aguda , Animales , Conducta Animal/fisiología , Enfermedad Crónica , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/genética , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/metabolismo , Neuronas Dopaminérgicas/metabolismo , Expresión Génica , Histamina/administración & dosificación , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Núcleo Accumbens/fisiopatología , Pruebas de Farmacogenómica , Cloruro de Picrilo/administración & dosificación , Prurito/genética , Tirosina 3-Monooxigenasa/genéticaRESUMEN
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by pruritic and eczematous skin lesions. The skin of AD patients is generally in a dried condition. Therefore, it is important for AD patients to manage skin moisturization. In this study, we examined the effects of orally administered fermented barley extract P (FBEP), which is prepared from a supernatant of barley shochu distillery by-product, on stratum corneum (SC) hydration and transepidermal water loss (TEWL) in AD-like lesions induced in hairless mice using 2,4,6-trinitrochlorobenzene. Oral administration of FBEP increased SC hydration and decreased TEWL in the dorsal skin of this mouse model. Further fractionation of FBEP showed that a pyroglutamyl pentapeptide, pEQPFP comprising all -L-form amino acids, is responsible for these activities. These results suggested that this pyroglutamyl pentapeptide may serve as a modality for the treatment of AD.
Asunto(s)
Mezclas Complejas/farmacología , Dermatitis Atópica/tratamiento farmacológico , Epidermis/efectos de los fármacos , Hordeum/química , Hipodermoclisis/métodos , Oligopéptidos/farmacología , Ácido Pirrolidona Carboxílico/análogos & derivados , Animales , Mezclas Complejas/aislamiento & purificación , Dermatitis Atópica/inducido químicamente , Dermatitis Atópica/patología , Modelos Animales de Enfermedad , Epidermis/patología , Fermentación , Masculino , Ratones , Ratones Pelados , Oligopéptidos/aislamiento & purificación , Cloruro de Picrilo/administración & dosificación , Ácido Pirrolidona Carboxílico/aislamiento & purificación , Ácido Pirrolidona Carboxílico/farmacología , Resultado del TratamientoRESUMEN
Allergic disease is one of the most important and common health problems worldwide. We have previously demonstrated that a fig leaf-derived lactic acid bacterium Lactobacillus (Lb.) paracasei IJH-SONE68 produces a novel exopolysaccharide (EPS). Furthermore, we have shown that the EPS inhibits the catalytic activity of hyaluronidase (EC 3.2.1.36) promoting inflammatory reactions. To evaluate the anti-allergy and anti-inflammatory effects of the EPS, in the present study, we employed the picryl-chloride-induced delayed-type (type IV) allergy model mice, which is used to evaluate the contact dermatitis. Oral administration of the EPS was observed to reduce the ear swelling in the model mice. We also observed that the overexpression of ear interleukin-4 (T helper (Th) 2 cytokine) mRNA and the increase in serum immunoglobulin E (IgE) are repressed. However, the expression of interferon-γ (Th1 cytokine) was not accelerated in all of the allergen-challenged model mice. The improvement may be responsible for the Th2 downregulation rather than the Th1 upregulation. In addition, the symptom of immediate-type (type I) allergy model mice was improved by oral administration of the IJH-SONE68 cell (data not shown). We can conclude that the IJH-SONE68-derived EPS is useful to improve the type I and IV allergies including atopic dermatitis.
Asunto(s)
Antialérgicos/farmacología , Antiinflamatorios/farmacología , Dermatitis por Contacto/prevención & control , Fármacos Dermatológicos/farmacología , Lacticaseibacillus paracasei/química , Polisacáridos Bacterianos/farmacología , Administración Oral , Alérgenos/inmunología , Alérgenos/metabolismo , Animales , Antialérgicos/aislamiento & purificación , Antiinflamatorios/aislamiento & purificación , Dermatitis por Contacto/etiología , Dermatitis por Contacto/inmunología , Dermatitis por Contacto/patología , Fármacos Dermatológicos/aislamiento & purificación , Modelos Animales de Enfermedad , Oído , Expresión Génica/efectos de los fármacos , Hialuronoglucosaminidasa/antagonistas & inhibidores , Hialuronoglucosaminidasa/inmunología , Hialuronoglucosaminidasa/metabolismo , Inmunoglobulina E/sangre , Inmunoglobulina E/genética , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-4/genética , Interleucina-4/inmunología , Lacticaseibacillus paracasei/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Cloruro de Picrilo/administración & dosificación , Polisacáridos Bacterianos/aislamiento & purificación , Células TH1/efectos de los fármacos , Células TH1/inmunología , Células Th2/efectos de los fármacos , Células Th2/inmunologíaRESUMEN
BACKGROUND: Application of haptens to the skin induces release of immune stimulatory ATP into the extracellular space. This "danger" signal can be converted to immunosuppressive adenosine (ADO) by the action of the ectonucleotidases CD39 and CD73, expressed by skin and immune cells. Thus, the expression and regulation of CD73 by skin derived cells may have crucial influence on the outcome of contact hypersensitivity (CHS) reactions. OBJECTIVE: To investigate the role of CD73 expression during 2,4,6-trinitrochlorobenzene (TNCB) induced CHS reactions. METHODS: Wild type (wt) and CD73 deficient mice were subjected to TNCB induced CHS. In the different mouse strains the resulting ear swelling reaction was recorded along with a detailed phenotypic analysis of the skin migrating subsets of dendritic cells (DC). RESULTS: In CD73 deficient animals the motility of DC was higher as compared to wt animals and in particular after sensitization we found increased migration of Langerin+ DC from skin to draining lymph nodes (LN). In the TNCB model this led to a stronger sensitization as indicated by increased frequency of interferon-γ producing T cells in the LN and an increased ear thickness after challenge. CONCLUSION: CD73 derived ADO production slows down migration of Langerin+ DC from skin to LN. This may be a crucial mechanism to avoid over boarding immune reactions against haptens.
Asunto(s)
5'-Nucleotidasa/metabolismo , Movimiento Celular/inmunología , Células Dendríticas/inmunología , Dermatitis Alérgica por Contacto/inmunología , Piel/citología , 5'-Nucleotidasa/genética , 5'-Nucleotidasa/inmunología , Adenosina/inmunología , Adenosina/metabolismo , Adenosina Trifosfato/inmunología , Adenosina Trifosfato/metabolismo , Animales , Antígenos de Superficie/metabolismo , Células Cultivadas , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Proteínas Ligadas a GPI/genética , Proteínas Ligadas a GPI/inmunología , Proteínas Ligadas a GPI/metabolismo , Haptenos/administración & dosificación , Haptenos/inmunología , Interferón gamma/metabolismo , Lectinas Tipo C/metabolismo , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Lectinas de Unión a Manosa/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Cloruro de Picrilo/administración & dosificación , Cloruro de Picrilo/inmunología , Piel/inmunología , Piel/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismoAsunto(s)
Movimiento Celular/efectos de los fármacos , Células Dendríticas/efectos de los fármacos , Dermatitis Alérgica por Contacto/prevención & control , Dinitrofluorobenceno/administración & dosificación , Haptenos , Inmunosupresores/administración & dosificación , Oxazolona/administración & dosificación , Cloruro de Picrilo/administración & dosificación , Administración Cutánea , Animales , Células Dendríticas/inmunología , Dermatitis Alérgica por Contacto/inmunología , Dinitrofluorobenceno/inmunología , Modelos Animales de Enfermedad , Inmunosupresores/inmunología , Ratones Endogámicos C57BL , Oxazolona/inmunología , Cloruro de Picrilo/inmunología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunologíaRESUMEN
BACKGROUND: Contact hypersensitivity (CHS) reaction induced by a topical application of hapten is a cell-mediated antigen-specific type of skin inflammation mediated by interaction of several subtypes of T cell subpopulations. Recently, it has been shown that antidepressant drugs inhibit CHS reaction, although the mechanism of this effect remains unknown. The aim of the present study was to investigate the effect of 2-week desipramine or fluoxetine administration on the CHS reaction induced by picryl chloride (PCL) application in B10.PL mice and in knock-out mice established on B10.PL background: TCRδ(-/-) mice lacking TCRγδ T lymphocytes; ß2m(-/-) mice lacking CD8(+) T lymphocytes and CD1d(-/-) mice lacking CD1d dependent natural killer T (NKT) lymphocytes. METHODS: B10.PL, TCRδ(-/-), ß2m(-/-) and CD1d(-/-) mice were divided into six groups: 1) vehicle-treated negative control group; 2) desipramine-treated negative control group; 3) fluoxetine-treated negative control group; 4) vehicle and PCL-treated group (positive control group); 5) desipramine and PCL-treated group; and 6) fluoxetine and PCL-treated group. CHS to PCL was tested by evaluation of ear swelling. Metabolic activity of spleen and lymph node cells were estimated by MTT test. RESULTS: The antidepressants significantly suppressed the CHS reaction in B10.PL mice: desipramine by 55% and fluoxetine by 42% compared to the positive control. This effect was even stronger in TCRδ(-/-) mice, in which fluoxetine reduced the ear swelling by 73% in comparison with the vehicle-treated positive control group. On the other hand, desipramine and fluoxetine did not inhibit CHS reaction in ß2m(-/-) and CD1d(-/-) mice. Moreover, PCL increased metabolic and/or proliferative activity of splenocytes in all four strains of mice whereas the antidepressants decreased this activity of splenocytes in B10.PL, TCRδ(-/-) and CD1d(-/-) mice. CONCLUSION: The results of the present study show that lack of CD8(+) T cells or NKT cells abolishes the immunosuppressive effect of antidepressant drugs on PCL-induced CHS reaction in mice. These results suggest that antidepressant drug-induced inhibition of CHS reaction is connected with their inhibitory effect on ability of CD8(+) T cells and NKT cells to induce and/or escalate CHS reaction. TCRγδ cells seem not to be involved in antidepressant-induced suppression of CHS.
Asunto(s)
Antidepresivos/administración & dosificación , Linfocitos T CD8-positivos/efectos de los fármacos , Dermatitis por Contacto/tratamiento farmacológico , Desipramina/administración & dosificación , Fluoxetina/administración & dosificación , Células T Asesinas Naturales/efectos de los fármacos , Animales , Antidepresivos/efectos adversos , Antígenos CD1d/genética , Linfocitos T CD8-positivos/fisiología , Dermatitis por Contacto/inmunología , Desipramina/efectos adversos , Fluoxetina/efectos adversos , Terapia de Inmunosupresión , Ratones , Ratones Endogámicos , Ratones Noqueados , Células T Asesinas Naturales/fisiología , Cloruro de Picrilo/administración & dosificación , Receptores de Antígenos de Linfocitos T gamma-delta/genética , Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Microglobulina beta-2/genéticaRESUMEN
The EphB/ephrinB receptor-ligand system is pivotal for the development of the embryonic vasculature and for angiogenesis in the adult organism. We observed that (i) the expression of ephrinB2 and ephrinB1 is up-regulated in capillaries during inflammation, that (ii) these ligands are localised on the luminal endothelial surface, and that (iii) they interact with the ephrinB-receptor EphB2 on monocyte/macrophages. This study delineates the impact of ephrinB-mediated reverse signalling on the integrity and proinflammatory differentiation of the endothelium. To this end, in vitro analyses with human cultured endothelial cells reveal that knockdown of ephrinB2 or ephrinB1 impairs monocyte transmigration through the endothelium. While ephrinB2 but not ephrinB1 interacts with PECAM-1 (CD31) in this context, reverse signalling by ephrinB1 but not ephrinB2 elicits a c-Jun N-terminal kinase (JNK)-dependent up-regulation of E-selectin expression. Furthermore, treatment of endothelial cells with soluble EphB2 receptor bodies or EphB2-overexpressing mouse myeloma cells links ephrinB2 to PECAM-1 and induces its Src-dependent phosphorylation while diminishing Src homology phosphotyrosyl phosphatase-2 (SHP-2) activity and increasing endothelial cell permeability. We conclude that extravasation of EphB2 positive leukocyte populations is facilitated by lowering the integrity of endothelial cell junctions and enhancing the pro-inflammatory phenotype of the endothelium through activation of ephrinB ligands.
Asunto(s)
Dermatitis Atópica/inmunología , Endotelio Vascular/fisiología , Efrina-B1/metabolismo , Efrina-B2/metabolismo , Monocitos/fisiología , Mieloma Múltiple/inmunología , Adulto , Animales , Permeabilidad Capilar , Diferenciación Celular/genética , Células Cultivadas , Dermatitis Atópica/inducido químicamente , Selectina E/genética , Selectina E/metabolismo , Efrina-B1/genética , Efrina-B2/genética , Regulación de la Expresión Génica , Humanos , Inflamación/inmunología , Uniones Intercelulares/genética , Ratones , Ratones Endogámicos , Ratones Transgénicos , Neoplasias Experimentales , Técnicas de Cultivo de Órganos , Cloruro de Picrilo/administración & dosificación , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/genética , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , ARN Interferente Pequeño/genética , Receptores de la Familia Eph/metabolismo , Transducción de Señal/genética , Migración Transendotelial y Transepitelial/genéticaRESUMEN
BACKGROUND: Thymic stromal lymphopoietin (TSLP) plays critical roles in the induction and exacerbation of allergic diseases. We tested various chemicals in the environment and found that xylene and 1,2,4-trimethylbenzene induced the production of TSLP in vivo. These findings prompted us to search for additional chemicals that induce TSLP production. In this study, we examined whether fatty acids could induce the production of TSLP in vivo and exacerbate allergic inflammation. METHODS: Various fatty acids and related compounds were painted on the ear lobes of mice and the amount of TSLP in the homogenate of ear lobe tissue was determined. The effects of nonanoic acid on allergic inflammation were also examined. RESULTS: Octanoic acid, nonanoic acid, and decanoic acid markedly induced TSLP production, while a medium-chain aldehyde and alcohol showed only weak activity. Nonanoic acid induced the production of TSLP with a maximum at 24 h. TSLP production was even observed in nonanoic acid-treated C3H/HeJ mice that lacked functional toll-like receptor 4. The aryl hydrocarbon receptor agonist ß-naphthoflavone did not induce TSLP production. Nonanoic acid promoted sensitization to ovalbumin, resulting in an enhancement in the cutaneous anaphylactic response. In addition, painting of nonanoic acid after the sensitization augmented picryl chloride-induced thickening of the ear, which was reversed in TSLP receptor-deficient mice. CONCLUSIONS: Nonanoic acid and certain fatty acids induced TSLP production, resulting in the exacerbation of allergic inflammation. We propose that TSLP-inducing chemical compounds such as nonanoic acid be recognized as chemical allergo-accelerators.
Asunto(s)
Citocinas/metabolismo , Ácidos Grasos/efectos adversos , Hipersensibilidad/inmunología , Inflamación/inmunología , Piel/patología , Alcoholes/administración & dosificación , Aldehídos/administración & dosificación , Animales , Citocinas/genética , Progresión de la Enfermedad , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Inmunoglobulinas/metabolismo , Inflamación/inducido químicamente , Ratones , Ratones Endogámicos C3H , Ratones Noqueados , Cloruro de Picrilo/administración & dosificación , Receptores de Hidrocarburo de Aril/agonistas , Receptores de Citocinas/genética , Receptores de Citocinas/metabolismo , Piel/efectos de los fármacos , Receptor Toll-Like 4/genética , beta-naftoflavona/administración & dosificación , Linfopoyetina del Estroma TímicoRESUMEN
We aimed to define whether vitamin E improves biochemical indices associated with symptoms of atopic dermatitis-like inflammation in NC/Nga mice. After picryl chloride (PC) application to their backs, changes in the content of thiobarbituric acid reactive substances (TBARS) and vitamin E, as well as the activity of antioxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase) were analyzed in the serum and skin of NC/Nga mice during a symptomatic cycle. The levels of inflammatory factors were also assessed, including IgE, cyclooxigenase-2 (COX-2), tumor necrosis factor (TNF-α) and nuclear factor-κB (NF-κB). When allergic dermatitis was induced by the application of PC to the skin of the mice, skin inflammation appeared 2 wk after PC application, with the peak severity of inflammation observed 5 wk after PC application. Subsequently, the animals recovered from the inflammation by 9 wk after PC application. The TBARS content in the skin and serum increased markedly when the symptoms were the most severe, and decreased to levels near those in control mice by 9 wk after PC application. The activities of SOD and GSHPx in the skin and serum were also positively correlated with symptomatic changes; however, no change in catalase activity was observed 5 wk after PC application. Conversely, vitamin E content decreased at the stage of peak severity. The levels of all inflammatory factors analyzed in this study were altered in a manner similar to other indices. Additionally, vitamin E treatment markedly inhibited these PC-induced alterations. On the basis of these results, it is expected that the observed alterations in biochemical indices, which reflect the symptomatic cycle, may be applicable to objective diagnosis and treatment for atopic dermatitis, and that vitamin E may improve the symptoms of AD.
Asunto(s)
Dermatitis Atópica/tratamiento farmacológico , Dermatitis Atópica/fisiopatología , Vitamina E/administración & dosificación , Animales , Catalasa/análisis , Catalasa/sangre , Ciclooxigenasa 2/análisis , Ciclooxigenasa 2/sangre , Dermatitis Atópica/inducido químicamente , Glutatión Peroxidasa/análisis , Glutatión Peroxidasa/sangre , Inmunoglobulina E/análisis , Inmunoglobulina E/sangre , Masculino , Ratones , FN-kappa B/análisis , FN-kappa B/sangre , Estrés Oxidativo , Cloruro de Picrilo/administración & dosificación , Piel/química , Piel/patología , Superóxido Dismutasa/análisis , Superóxido Dismutasa/sangre , Sustancias Reactivas al Ácido Tiobarbitúrico/análisis , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/sangre , Vitamina E/análisis , Vitamina E/sangreRESUMEN
Cytohesin-interacting protein (Cytip) is induced during dendritic cell (DC) maturation and in T cells upon activation. It has also been shown to be involved in the regulation of immune responses. Here, we evaluated the functional consequences of Cytip deficiency in DCs using Cytip knockout (KO) mice. No difference in DC subpopulations in the skin draining lymph nodes (LNs) was found between Cytip KO mice and their wild-type counterparts, excluding a role in DC development. To investigate the function of Cytip in DCs in vivo, we used 2,4,6-trinitrochlorobenzene (TNCB)-induced contact hypersensitivity (CHS) as a model system. In the sensitization as well as in the elicitation phase, DCs derived from Cytip KO mice induced an increased inflammatory reaction indicated by more pronounced ear swelling. Furthermore, IL-12 production was increased in Cytip KO bone marrow-derived DCs (BMDCs) after CpG stimulation. Additionally, Cytip-deficient DCs loaded with ovalbumin induced stronger proliferation of antigen-specific CD4(+) and CD8(+) T cells in vitro. Finally, migration of skin DCs was not altered after TNCB application due to Cytip deficiency. Taken together, these data suggest a suppressive function for Cytip in mouse DCs in limiting immune responses.
Asunto(s)
Proteínas Portadoras/inmunología , Células Dendríticas/inmunología , Dermatitis por Contacto/inmunología , Proteínas de la Membrana/inmunología , Linfocitos T/inmunología , Animales , Proteínas Portadoras/genética , Procesos de Crecimiento Celular/inmunología , Células Dendríticas/citología , Dermatitis por Contacto/patología , Citometría de Flujo , Antígenos de Histocompatibilidad Clase II/inmunología , Inflamación/inducido químicamente , Inflamación/inmunología , Inflamación/patología , Interleucina-12/genética , Interleucina-12/inmunología , Activación de Linfocitos/inmunología , Proteínas de la Membrana/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Cloruro de Picrilo/administración & dosificación , ARN Mensajero/química , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Piel/inmunología , Piel/patología , Linfocitos T/patologíaRESUMEN
Contact hypersensitivity (CHS) is thought to be associated mainly with the activation of T helper (Th) type 1 cells. However, evidence also suggests that Th type 2 cells (Th2) and cytokines play roles in the development of CHS in humans. The present study examines the Th2 response during the development of CHS in response to 2,4,6-trinitrochlorobenzene (TNCB) in GATA-3-transgenic (GATA-3 Tg) mice. GATA-3 Tg mice were immunized with 7% TNCB applied to abdominal shaved skin. Seven days later, the mice were challenged with 1% TNCB applied to the left ear. Ear swelling, cytokine production in the skin of the ear, and the levels of IgE, IgG1 and IgG2a were measured. Furthermore, we examined the effects of medical treatment on TNCB-induced contact dermatitis using this model. The ear-swelling responses of TNCB-sensitized/challenged GATA-3 Tg mice were significantly greater than those of similarly treated wild-type (WT) mice. The expression of both IL-5 and IL-13 in TNCB sensitized/challenged skin tissues and the IgE response after challenge were obviously increased in the GATA-3 Tg mice, whereas the expression of IFN-γ was identical in the challenged skin tissues of GATA-3-Tg and WT mice. When TNCB-sensitized GATA-3 Tg mice were treated with a high dose of tacrolimus, ear swelling was not significantly decreased, compared with the results in WT mice. These results suggest that GATA-3-induced Th2-dominant responses play a critical role in the pathogenesis of allergic types of dermatitis, such as atopic dermatitis, and may lead to useful new drug development in the future.
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Dermatitis Alérgica por Contacto/inmunología , Factor de Transcripción GATA3/metabolismo , Piel/metabolismo , Animales , Separación Celular , Células Cultivadas , Dermatitis Alérgica por Contacto/sangre , Dermatitis Alérgica por Contacto/tratamiento farmacológico , Modelos Animales de Enfermedad , Citometría de Flujo , Factor de Transcripción GATA3/genética , Factor de Transcripción GATA3/inmunología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Inmunización , Inmunoglobulina E/genética , Inmunoglobulina E/inmunología , Inmunoglobulina E/metabolismo , Interleucina-13/genética , Interleucina-13/inmunología , Interleucina-13/metabolismo , Interleucina-5/genética , Interleucina-5/inmunología , Interleucina-5/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Cloruro de Picrilo/administración & dosificación , Piel/efectos de los fármacos , Piel/inmunología , Piel/patología , Tacrolimus/administración & dosificación , Tacrolimus/efectos adversos , Células Th2/inmunología , Células Th2/trasplanteRESUMEN
A chymase inhibitor SUN13834 has been shown to improve skin condition in animal models for atopic dermatitis. In the present study, effective dosages of SUN13834 for atopic dermatitis patients were predicted by pharmacokinetic/pharmacodynamic (PK/PD) analyses of SUN13834 in NC/Nga mice, which spontaneously develop atopic dermatitis-like skin lesions. For the PK/PD analyses, we utilized the minimum effective plasma concentration of unbound SUN13834 in late-phase reaction of trinitrochlorobenzene (TNCB)-induced biphasic dermatitis in mice, based on the assumption that the minimum effective plasma concentrations are the same among the two animal models. In late-phase reaction of biphasic dermatitis, SUN13834 was most effective when its plasma concentration was highest at the elicitation, and the minimum effective plasma concentration of unbound SUN13834 at the elicitation was calculated to be 0.13-0.2 ng/mL. Oral administration of SUN13834 improved dermatitis in NC/Nga mice at 15 mg/kg (twice a day; bid) and 30 mg/kg (once a day; qd), but not at 60 mg/kg (every other day; eod). At the three dosages, the duration times over the plasma level of 0.13-0.2 ng/mL were 16.1-20.3, 10.7-12.2 and 7.8-8.8h, respectively, suggesting an importance of maintenance of the minimum effective plasma concentration for at least about 10-12h. The clinical effective dosage predicted in this paper is also discussed in relation to a recently conducted Phase 2a study.
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Azepinas/administración & dosificación , Quimasas/metabolismo , Dermatitis Atópica/tratamiento farmacológico , Inhibidores Enzimáticos/administración & dosificación , Piel/efectos de los fármacos , Administración Oral , Animales , Azepinas/farmacocinética , Ensayos Clínicos Fase II como Asunto , Dermatitis Atópica/inducido químicamente , Dermatitis Atópica/enzimología , Susceptibilidad a Enfermedades , Cálculo de Dosificación de Drogas , Inhibidores Enzimáticos/farmacocinética , Humanos , Ratones , Ratones Endogámicos , Cloruro de Picrilo/administración & dosificación , Piel/patologíaRESUMEN
We investigated the synergetic effects of glucocorticoid and histamine H1 receptor antagonists on an atopic dermatitis model. Hairless mice were used in this study and an atopic dermatitis model was made by repeated application of 2,4,6-trinitrochlorobenzene. The effects of glucocorticoid, histamine H1 receptor antagonists, and the simultaneous use of these drugs were investigated by measuring scratching behavior, skin symptoms and nerve growth factor (NGF) in the skin. Topical application of prednisolone significantly inhibited scratching behavior, skin symptoms and NGF contents in the skin by repeated application. Olopatadine also showed a significant effect on scratching behavior and NGF contents in the skin, whereas chlorpheniramine showed no significant inhibitory effect on these indices. Furthermore, the combined use of prednisolone and olopatadine potentiated the inhibition of scratching behavior, skin symptoms, and NGF in the skin. From these findings, olopatadine potentiated the inhibitory effect of prednisolone on the symptoms of atopic dermatitis by inhibiting NGF.
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Dermatitis Atópica/tratamiento farmacológico , Dibenzoxepinas/uso terapéutico , Glucocorticoides/uso terapéutico , Antagonistas de los Receptores Histamínicos H1/uso terapéutico , Prednisolona/uso terapéutico , Alérgenos/administración & dosificación , Alérgenos/inmunología , Animales , Antipruriginosos/uso terapéutico , Conducta Animal/efectos de los fármacos , Dermatitis Atópica/inmunología , Dermatitis Atópica/metabolismo , Dermatitis Atópica/patología , Dibenzoxepinas/administración & dosificación , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Sinergismo Farmacológico , Glucocorticoides/administración & dosificación , Antagonistas de los Receptores Histamínicos H1/administración & dosificación , Masculino , Ratones , Ratones Pelados , Factor de Crecimiento Nervioso/metabolismo , Clorhidrato de Olopatadina , Cloruro de Picrilo/administración & dosificación , Cloruro de Picrilo/inmunología , Prednisolona/administración & dosificación , Prurito/tratamiento farmacológico , Prurito/inmunología , Índice de Severidad de la Enfermedad , Piel/efectos de los fármacos , Piel/inmunología , Piel/metabolismo , Piel/patología , Factores de TiempoRESUMEN
We examined the effects single and combined administration of fermented barley extract P (FBEP), prepared from barley-shochu distillery by-products, and gamma-aminobutyric acid (GABA) on the development of atopic dermatitis (AD)-like skin lesions in NC/Nga mice. Single administration of FBEP and GABA dose-dependently reduced the development of AD-like skin lesions in mice. GABA reduced the development of AD-like skin lesions by suppressing serum immunoglobulin E (IgE) and splenocyte interleukin (IL)-4 production, while FBEP reduced skin lesions without affecting the IgE or cytokine production. However, in mice with induced AD-like skin lesions, combined administration of FBEP and GABA decreased serum IgE levels and splenic cell IL-4 production, and increased splenic cell interferon-gamma production. These results suggest that combined administration of FBEP and GABA alleviated AD-like skin lesions in the NC/Nga mice by adjusting the Th1/Th2 balance to a Th1-predominant immune response.
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Dermatitis Atópica/prevención & control , Fermentación , Hordeum/química , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología , Ácido gamma-Aminobutírico/administración & dosificación , Ácido gamma-Aminobutírico/farmacología , Animales , Peso Corporal/efectos de los fármacos , Citocinas/metabolismo , Dermatitis Atópica/sangre , Dermatitis Atópica/inducido químicamente , Dermatitis Atópica/fisiopatología , Femenino , Hordeum/metabolismo , Inmunoglobulina E/sangre , Ratones , Cloruro de Picrilo/administración & dosificación , Cloruro de Picrilo/farmacología , Extractos Vegetales/metabolismoRESUMEN
Previously, we indicated that athymic BALB/c-nu/nu (nude) mice that had been repeatedly treated with 2,4,6-trinitrochlorobenzene (TNCB) failed to exhibit chronic scratching behavior in spite of the accumulation of dermal mast cells in the lesion. The mice also failed to produce specific IgE or potent dermatitis. In the present study, therefore, we aimed to examine the role of IgE and repeated hapten treatment in the induction of scratching behavior and dermatitis using nude mice and trinitrophenol (TNP)-specific IgE-transgenic mice. The ears of nude mice were treated with TNCB 6 times at intervals of 48 h, and TNP-specific IgE was administered to the mice intravenously before the sixth TNCB treatment. The nude mice that had been supplemented with IgE exhibited a persistent increase in scratching behavior and continuous degranulation of mast cells. Furthermore, a potent immediate ear swelling was induced, although no biphasic dermatitis pattern was observed. On the other hand, the IgE-transgenic mice failed to exhibit persistent increases in scratching behavior after a single TNCB treatment, although biphasic ear swelling was observed. These results indicate that specific IgE plays an essential role in the induction of persistent increases in scratching behavior and continuous degranulation of mast cells. Furthermore, repeated challenge with the hapten also plays an important role in persistent increases in scratching behavior through accumulation and continuous activation of mast cells.
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Dermatitis Alérgica por Contacto/inmunología , Inmunoglobulina E/inmunología , Prurito/inmunología , Animales , Dermatitis Alérgica por Contacto/fisiopatología , Modelos Animales de Enfermedad , Femenino , Haptenos/inmunología , Inmunoglobulina E/administración & dosificación , Mastocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Ratones Transgénicos , Picratos/inmunología , Cloruro de Picrilo/administración & dosificación , Cloruro de Picrilo/inmunología , Prurito/etiologíaRESUMEN
The recently described cytokines IL-19, IL-20, and IL-24 share structural homology with IL-10 and are therefore classified as members of the IL-10 family of cytokines. Although it has long been speculated that signaling by their heterodimeric receptor complexes (IL-20R1/IL-20R2 and IL-22R/IL-20R2) influences immunological processes, the target cells for this group of cytokines are still unclear. By generating a knockout mouse strain deficient for the common IL-20R beta-chain (IL-20R2), we show that IFN-gamma and IL-2 secretion is significantly elevated after stimulation of IL-20R2-/--deficient CD8 and CD4 T cells with Con A or anti-CD3/CD28 in vitro. IL-10 secretion by activated IL-20R2-/- CD4 cells was diminished. Consistent with our in vitro results, significantly more Ag-specific CD8 IFN-gamma+ and CD4 IFN-gamma+ T cells developed to locally applied DNA vaccines in IL-20R2-deficient mice. In a T cell-dependent model of contact hypersensitivity, IL-20R2 knockout mice were more sensitive to the contact allergen trinitro-chloro-benzene. Thus, IL-20R2 signaling directly regulates CD8 and CD4 T cell answers in vitro and in vivo. For the first time, we provide evidence that IL-19, IL-20, and IL-24 are part of a signaling network that normally down-modulates T cell responses in mice.
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Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Regulación hacia Abajo/inmunología , Epítopos de Linfocito T/inmunología , Receptores de Interleucina/fisiología , Transducción de Señal/inmunología , Alérgenos/administración & dosificación , Alérgenos/inmunología , Animales , Células Cultivadas , Técnicas de Cocultivo , Dermatitis por Contacto/genética , Dermatitis por Contacto/inmunología , Regulación hacia Abajo/genética , Femenino , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Cloruro de Picrilo/administración & dosificación , Cloruro de Picrilo/inmunología , Receptores de Interleucina/deficiencia , Receptores de Interleucina/genética , Transducción de Señal/genética , Vacunas de ADN/administración & dosificación , Vacunas de ADN/inmunologíaRESUMEN
In the present paper, the effect of roxithromycin on delayed-type hypersensitivity (DTH) was evaluated. Roxithromycin had no effect on sheep red blood cells (SRBC)-induced food pad swelling when orally administered in induction phase, whereas it suppressed the SRBC-induced DTH reaction and 2,4,6-Trinitrochlorobenzene (TNCB)-induced contact hypersensitivity (CHS) significantly when administered to mice in effector phase. For the sustained-CHS model induced by multi-challenge with TNCB, roxithromycin also inhibited the ear swelling when exposed to mice in three effector phases while showed no inhibitory effect on CHS by continuous treatment. Administration of this antibiotic in effector phase also down-regulated the MMP-9 activity and the higher in vitro survival of splenocytes from SRBC-challenged mice. Furthermore, this drug inhibited the gene expression of T-helper type 1 (Th1) cytokines such as IL-2 and IFN-gamma of lymph node cells from mice immuned by TNCB or of Con A-stimulated spleen cells. In addition, roxithromycin administered in vivo decreased the concanavalin A (Con A)-induced splenocyte proliferation without affecting the cell survival in vitro. These results suggest that roxithromycin might alleviate DTH reaction at least by suppressing the function and survival of effector T cells.
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Hipersensibilidad Tardía/tratamiento farmacológico , Hipersensibilidad Tardía/inmunología , Inmunosupresores/administración & dosificación , Roxitromicina/administración & dosificación , Administración Oral , Animales , Dermatitis por Contacto/tratamiento farmacológico , Dermatitis por Contacto/inmunología , Oído Externo , Edema/inducido químicamente , Edema/inmunología , Eritrocitos/inmunología , Femenino , Miembro Posterior , Inmunidad Celular/efectos de los fármacos , Inmunosupresores/uso terapéutico , Ratones , Ratones Endogámicos ICR , Cloruro de Picrilo/administración & dosificación , Roxitromicina/uso terapéutico , OvinosRESUMEN
In cutaneous contact sensitivity there is an early elicited innate cascade of complement, mast cells, and platelets activated via IgM Abs. This response is required to initiate the elicitation of acquired classical contact sensitivity by leading to local recruitment of effector T cells. We recently performed in vivo experiments showing that collaboration is required between innate-like invariant Valpha14+ NKT cells (iNKT) and the innate-like B-1 B cell subset to induce this initiation process. Contact sensitization triggers iNKT cells to produce IL-4 to coactivate the B-1 cells along with specific Ag for production of the initiating IgM Abs. We now describe in vitro collaboration of iNKT and B-1 cells. Normal peritoneal B-1 cells, incubated in vitro with soluble Ag, and with 1-h in vivo immune iNKT cells producing IL-4, are activated to mediate the contact sensitivity-initiation cascade. The three components of this process can be activated by different Ag. Thus, 1-h iNKT cell activation, B-1 cell stimulation, and generation of immune effector T cells can be induced by sensitization with three different Ag to respectively generate IL-4 and Ag-specific IgM Abs, to recruit the Ag-specific effector T cells. These findings have relevance to allergic and autoimmune diseases in which infections can trigger exacerbation of T cell responses to allergens or to autoantigens.