RESUMEN
Interleukin (IL)-17A is a well-described mediator of bone resorption in inflammatory diseases, and postmenopausal osteoporosis is associated with increased serum levels of IL-17A. Ovariectomy (OVX) can be used as a model to study bone loss induced by estrogen deficiency and the role of IL-17A in osteoporosis development has previously been investigated using various methods to inhibit IL-17A signaling in this model. However, the studies show opposing results. While some publications reported IL-17A as a mediator of OVX-induced osteoporosis, others found a bone-protective role for IL-17 receptor signaling. In this study, we provide an explanation for the discrepancies in previous literature and show for the first time that loss of IL-17A has differential effects on OVX-induced osteoporosis; with IL-17A being important for cortical but not trabecular bone loss. Interestingly, the decrease in trabecular bone after OVX in IL-17A knock-out mice, was accompanied by increased adipogenesis depicted by elevated leptin levels. Additionally, the bone marrow adipose tissue expanded, and the bone-turnover decreased in ovariectomized mice lacking IL-17A compared to ovariectomized WT mice. Our results increase the understanding of how IL-17A signaling influences bone remodeling in the different bone compartments, which is of importance for the development of new treatments of post-menopausal osteoporosis.
Asunto(s)
Interleucina-17/fisiología , Osteoporosis/fisiopatología , Absorciometría de Fotón , Animales , Hueso Esponjoso/patología , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Fémur/diagnóstico por imagen , Fémur/patología , Humanos , Ratones , Ratones Noqueados , Osteogénesis/efectos de los fármacos , Osteoporosis/diagnóstico por imagen , Osteoporosis/etiología , Osteoporosis/patología , Osteoporosis Posmenopáusica/etiología , Osteoporosis Posmenopáusica/fisiopatología , Ovariectomía/efectos adversos , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Interleucina-17/fisiología , Microtomografía por Rayos XRESUMEN
Dectin-1 (gene symbol: Clec7a) is a receptor for ß-glucans that play an important role for the host defense against fungi. Recently, we showed that Clec7a-/- mice are resistant against dextran sodium sulfate (DSS)-induced colitis because of regulatory T-cell population expansion in the colon. The regulatory T-cell expansion is caused by expansion of commensal Lactobacillus murinus whose growth is suppressed by an antimicrobial protein, calprotectin S100A8/A9. In this report, we showed that S100A8 was mainly produced by mouse colonic epithelial cells. S100A8 was not induced directly by Dectin-1 but by Dectin-1-induced cytokines, especially interleukin-17F (IL-17F), that were produced by several types of innate immune cells including CD11c+/CD11b+ myeloid cells in colonic lamina propria. S100A8/A9 heterodimer preferentially suppressed the growth of L. murinus that was increased in both Clec7a-/- and Il17f-/- mice. Furthermore, similar expansion of L. murinus and DSS-colitis resistance were observed in mice fed with ß-glucan-free food. These observations suggest that food-derived ß-glucans control the specific commensal microbiota via the Dectin-1-IL-17F-calprotectin axis to maintain the intestinal homeostasis.
Asunto(s)
Colitis/inmunología , Colon/inmunología , Microbioma Gastrointestinal/fisiología , Mucosa Intestinal/fisiología , Lactobacillus/fisiología , Complejo de Antígeno L1 de Leucocito/metabolismo , Células Mieloides/fisiología , beta-Glucanos/administración & dosificación , Animales , Antiinfecciosos/metabolismo , Calgranulina A/metabolismo , Colitis/inducido químicamente , Colitis/genética , Alimentos , Interacciones Microbiota-Huesped , Interleucina-17/genética , Interleucina-17/metabolismo , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Complejo de Antígeno L1 de Leucocito/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , beta-Glucanos/metabolismoRESUMEN
Uveitis (intraocular inflammation) is a leading cause of loss of vision. Although its aetiology is largely speculative, it is thought to arise from complex genetic-environmental interactions that break immune tolerance to generate eye-specific autoreactive T cells. Experimental autoimmune uveitis (EAU), induced by immunization with the ocular antigen, interphotoreceptor retinoid binding protein (IRBP), in combination with mycobacteria-containing complete Freund's adjuvant (CFA), has many clinical and histopathological features of human posterior uveitis. Studies in EAU have focused on defining pathogenic CD4+ T cell effector responses, such as those of T helper type 17 (Th17) cells, but the innate receptor pathways precipitating development of autoreactive, eye-specific T cells remain poorly defined. In this study, we found that fungal-derived antigens possess autoimmune uveitis-promoting function akin to CFA in conventional EAU. The capacity of commensal fungi such as Candida albicans or Saccharomyces cerevisae to promote IRBP-triggered EAU was mediated by Card9. Because Card9 is an essential signalling molecule of a subgroup of C-type lectin receptors (CLRs) important in host defence, we evaluated further the proximal Card9-activating CLRs. Using single receptor-deficient mice we identified Dectin-2, but not Mincle or Dectin-1, as a predominant mediator of fungal-promoted uveitis. Conversely, Dectin-2 activation by α-mannan reproduced the uveitic phenotype of EAU sufficiently, in a process mediated by the Card9-coupled signalling axis and interleukin (IL)-17 production. Taken together, this report relates the potential of the Dectin-2/Card9-coupled pathway in ocular autoimmunity. Not only does it contribute to understanding of how innate immune receptors orchestrate T cell-mediated autoimmunity, it also reveals a previously unappreciated ability of fungal-derived signals to promote autoimmunity.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Proteínas Adaptadoras de Señalización CARD/inmunología , Candida albicans/inmunología , Candidiasis/inmunología , Lectinas Tipo C/inmunología , Saccharomyces cerevisiae/inmunología , Uveítis/inmunología , Animales , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/patología , Proteínas Adaptadoras de Señalización CARD/genética , Candidiasis/inducido químicamente , Candidiasis/patología , Proteínas del Ojo/toxicidad , Lectinas Tipo C/genética , Ratones , Ratones Mutantes , Proteínas de Unión al Retinol/toxicidad , Células Th17/inmunología , Células Th17/patología , Uveítis/inducido químicamente , Uveítis/genética , Uveítis/patologíaRESUMEN
The role of interleukin-17A (IL-17A) in host defense against Legionella pneumophila remains elusive. To address this issue, we used Il17a(-/-), Il17f(-/-), and Il17a/Il17f(-/-) mice on a C57Bl/6 (non-permissive) background and IL-17 neutralizing Abs in mice on an A/J (permissive) background. Higher bacterial (L. pneumophila) counts in the lung and blood along with reduced neutrophil recruitment were detected in Il17a(-/-), but not Il17f(-/-), mice. We found that neutrophils produce IL-17A homodimer (IL-17A) during L. pneumophila infection, and hematopoietic cell-derived IL-17A is known to be important for bacterial clearance. Thus, intratracheal administration of wild-type neutrophils or recombinant IL-17A restored bacterial clearance and neutrophil recruitment in Il17a(-/-) mice. Furthermore, neutrophil-depleted Rag2(-/-) and Rag2/Il-2rγ(-/-) mice exhibited increased bacterial burden, reduced neutrophil influx and IL-17A production in the lung. Recombinant IFN-γ administration in Il17a(-/-) mice augmented bacterial elimination, whereas IL-17A administration in Ifnγ(-/-) mice did not augment bacterial clearance. IFN-γ is produced by T cells, but not neutrophils or macrophages, suggesting that neutrophil-derived IL-17A induces IFN-γ in a paracrine fashion. Human pneumonic lungs and human neutrophils challenged with L. pneumophila exhibited increased numbers of IL-17A producing cells. These findings display a novel function of neutrophil-derived IL-17A in antibacterial defense via the induction of IFN-γ in a paracrine manner.
Asunto(s)
Interferón gamma/metabolismo , Interleucina-17/metabolismo , Legionella pneumophila/inmunología , Enfermedad de los Legionarios/inmunología , Pulmón/inmunología , Neutrófilos/inmunología , Animales , Anticuerpos Neutralizantes/administración & dosificación , Carga Bacteriana , Células Cultivadas , Dimerización , Femenino , Humanos , Interferón gamma/genética , Interleucina-17/genética , Interleucina-17/inmunología , Pulmón/microbiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Infiltración Neutrófila , Neutrófilos/microbiología , Comunicación ParacrinaRESUMEN
Hematopoietic stem cell transplantation (HSCT) efficacy is limited by numerous pulmonary complications. We developed a model of syngeneic bone marrow transplantion (BMT) followed by infection with murine gamma herpesvirus-68 that results in pneumonitis and fibrosis and mimics human "noninfectious" HSCT complications. BMT mice experience increased early lytic replication, but establish viral latency by 21 days post infection. CD4 T cells in BMT mice are skewed toward interleukin (IL)-17A rather than interferon (IFN)-γ production. Transplantation of bone marrow from Il-17a(-/-) donors or treatment with anti-IL-17A neutralization antibodies at late stages attenuates pneumonitis and fibrosis in infected BMT mice, suggesting that hematopoietic-derived IL-17A is essential for development of pathology. IL-17A directly influences activation and extracellular matrix production by lung mesenchymal cells. Lung CD11c+ cells of BMT mice secrete more transforming growth factor beta-ß1, and pro-TH17 mRNAs for IL-23 and IL-6, and less TH1-promoting cytokine mRNA for IFN-γ but slightly more IL-12 mRNA in response to viral infection. Adoptive transfer of non-BMT lung CD11c-enriched cells restores robust TH1 response and suppresses aberrant TH17 response in BMT mice to improve lung pathology. Our data suggest that "noninfectious" HSCT lung complications may reflect preceding viral infections and demonstrate that IL-17A neutralization may offer therapeutic advantage even after disease onset.
Asunto(s)
Células Presentadoras de Antígenos/inmunología , Trasplante de Médula Ósea , Infecciones por Herpesviridae/inmunología , Pulmón/patología , Neumonía/inmunología , Complicaciones Posoperatorias/inmunología , Rhadinovirus/fisiología , Células Th17/inmunología , Animales , Anticuerpos Neutralizantes/administración & dosificación , Células Cultivadas , Modelos Animales de Enfermedad , Fibrosis , Humanos , Interleucina-17/genética , Interleucina-17/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neumonía/etiología , Neumonía/prevención & control , Complicaciones Posoperatorias/prevención & control , Células Th17/virología , Latencia del Virus , Replicación ViralRESUMEN
Primary aldosteronism due to unilateral aldosterone-producing adenoma (APA) is a surgically curable form of hypertension. Bilateral APA can also be surgically curable in theory but few successful cases can be found in the literature. It has been reported that even using successful adrenal venous sampling (AVS) via bilateral adrenal central veins, it is extremely difficult to differentiate bilateral APA from bilateral idiopathic hyperaldosteronism (IHA) harbouring computed tomography (CT)-detectable bilateral adrenocortical nodules. We report a case of bilateral APA diagnosed by segmental AVS (S-AVS) and blood sampling via intra-adrenal first-degree tributary veins to localize the sites of intra-adrenal hormone production. A 36-year-old man with marked long-standing hypertension was referred to us with a clinical diagnosis of bilateral APA. He had typical clinical and laboratory profiles of marked hypertension, hypokalaemia, elevated plasma aldosterone concentration (PAC) of 45.1 ng dl(-1) and aldosterone renin activity ratio of 90.2 (ng dl(-1) per ng ml(-1 )h(-1)), which was still high after 50 mg-captopril loading. CT revealed bilateral adrenocortical tumours of 10 and 12 mm in diameter on the right and left sides, respectively. S-AVS confirmed excess aldosterone secretion from a tumour segment vein and suppressed secretion from a non-tumour segment vein bilaterally, leading to the diagnosis of bilateral APA. The patient underwent simultaneous bilateral sparing adrenalectomy. Histopathological analysis of the resected adrenals together with decreased blood pressure and PAC of 5.2 ng dl(-1) confirmed the removal of bilateral APA. S-AVS was reliable to differentiate bilateral APA from IHA by direct evaluation of intra-adrenal hormone production.
Asunto(s)
Neoplasias de la Corteza Suprarrenal/diagnóstico , Neoplasias de la Corteza Suprarrenal/cirugía , Adrenalectomía/métodos , Adenoma Corticosuprarrenal/diagnóstico , Adenoma Corticosuprarrenal/cirugía , Aldosterona/sangre , Biomarcadores de Tumor/sangre , Recolección de Muestras de Sangre/métodos , Tratamientos Conservadores del Órgano , Neoplasias de la Corteza Suprarrenal/sangre , Neoplasias de la Corteza Suprarrenal/metabolismo , Adenoma Corticosuprarrenal/sangre , Adenoma Corticosuprarrenal/metabolismo , Adulto , Aldosterona/metabolismo , Biomarcadores de Tumor/metabolismo , Biopsia , Diagnóstico Diferencial , Humanos , Inmunohistoquímica , Masculino , Valor Predictivo de las Pruebas , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , VenasRESUMEN
Approximately 10% of cases of hypertension in Japan are caused by primary aldosteronism (PA), amounting to about 4 million patients in total. Primary aldosteronism due to unilateral aldosterone hypersecretion is potentially curable by adrenalectomy. The clinical benefits of identifying and treating PA have been reported internationally, but its cost-effectiveness is unclear. We examined whether diagnosing and treating hidden PA in hypertensive population was cost-effective compared with suboptimal treatment. Our hypothetical patient was a 50-year-old man diagnosed with stage I-III hypertension. We established a Markov decision model based on plausible clinical pathways and prognoses of PA. We applied cost-effectiveness analysis comparing a comprehensive diagnostic strategy for PA (measurement of plasma aldosterone/renin ratio, 2 loading tests, imaging, and selective adrenal venous sampling) with a suboptimal strategy to manage hypertension by medication unless the typical signs of PA or other complication were manifest. Outcome measures were expected costs, expected effectiveness, and incremental cost-effectiveness ratio. The robustness of the findings was established by one-way and scenario sensitivity analyses. The comprehensive PA diagnostic strategy increased the expected costs by 64 004 JPY and expected life-years by 0.013 compared with standard treatment. The incremental cost-effectiveness ratio for the diagnosis of PA was 4 923 385 JPY per year. Our findings were sensitive to the outcomes of screening and treatment, and the costs of continuous or periodic medication for hypertension and the treatment of stroke and its complications.
Asunto(s)
Análisis Costo-Beneficio , Hiperaldosteronismo/diagnóstico , Hiperaldosteronismo/terapia , Humanos , Hiperaldosteronismo/economía , Japón , Masculino , Cadenas de Markov , Persona de Mediana EdadRESUMEN
Neuregulin-1 (NRG1) is a well-recognized risk gene for schizophrenia and is often implicated in the neurodevelopmental hypothesis of this illness. Alternative splicing and proteolytic processing of the NRG1 gene produce more than 30 structural variants; however, the neuropathological roles of individual variants remain to be characterized. On the basis of the neurodevelopmental hypothesis of schizophrenia, we administered eNRG1 (0.1~1.0 µg/g), a core epidermal growth factor-like (EGF) domain common for all splicing NRG1 variants, to neonatal mice and compared their behavioral performance with mice challenged with a full mature form of type 1 NRG1 variant. During the neonatal stage, recombinant eNRG1 protein administrated from the periphery passed the blood-brain barrier and activated its receptor (ErbB4) in the brain. In adults, the mice receiving the highest dose exhibited lower locomotor activity and deficits in prepulse inhibition and tonedependent fear learning, although the hearing reduction of the eNRG1-treated mice may explain these behavioral deficits. Neonatal eNRG1 treatment also significantly potentiated MK-801-driven locomotor activity in an eNRG1 dose-dependent manner. In parallel eNRG1 treatment enhanced MK-801-driven c-Fos induction and decreased immunoreactivity for NMDA receptor subunits in adult brain. In contrast, mice that had been treated with the same molar dose of a full mature form of type 1 NRG1 as neonates did not exhibit hypersensitivity to MK-801. However, both animal models exhibited similar hypersensitivity to methamphetamine. Collectively, our findings suggest that aberrant peripheral NRG1 signals during neurodevelopment alter later behavioral traits and auditory functions in the NRG1 subtype-dependent manner.
Asunto(s)
Maleato de Dizocilpina/farmacología , Neurregulina-1/uso terapéutico , Esquizofrenia/tratamiento farmacológico , Animales , Animales Recién Nacidos , Encéfalo/metabolismo , Maleato de Dizocilpina/uso terapéutico , Evaluación Preclínica de Medicamentos , Femenino , Masculino , Ratones Endogámicos C57BL , Actividad Motora/efectos de los fármacos , Neurregulina-1/farmacocinética , Subunidades de Proteína/antagonistas & inhibidores , Subunidades de Proteína/metabolismo , Proteínas Proto-Oncogénicas c-fos/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Receptores de N-Metil-D-Aspartato/metabolismo , Distribución TisularRESUMEN
Induction of mucosal immunoglobulin-A (IgA) capable of providing a first line of defense against bacterial and viral pathogens remains a major goal of needle-free vaccines given via mucosal routes. Innate immune cells are known to play a central role in induction of IgA responses by mucosal vaccines, but the relative contribution of myeloid cell subsets to these responses has not firmly been established. Using an in vivo model of sublingual vaccination with Bacillus anthracis edema toxin (EdTx) as adjuvant, we examined the role of myeloid cell subsets for mucosal secretory IgA responses. Sublingual immunization of wild-type mice resulted in a transient increase of neutrophils in sublingual tissues and cervical lymph nodes. These mice later developed Ag-specific serum IgG responses, but not serum or mucosal IgA. Interestingly, EdTx failed to increase neutrophils in sublingual tissues and cervical lymph nodes of IKKß(ΔMye) mice, and these mice developed IgA responses. Partial depletion of neutrophils before immunization of wild-type mice allowed the development of both mucosal and serum IgA responses. Finally, co-culture of B cells with neutrophils from either wild-type or IKKß(ΔMye) mice suppressed secretion of IgA, but not IgM or IgG. These results identify a new role for neutrophils as negative regulators of IgA responses.
Asunto(s)
Inmunidad Mucosa , Inmunoglobulina A Secretora/inmunología , Membrana Mucosa/inmunología , Neutrófilos/inmunología , Administración Sublingual , Animales , Formación de Anticuerpos , Antígenos Bacterianos/inmunología , Linfocitos B/inmunología , Linfocitos B/metabolismo , Toxinas Bacterianas/inmunología , Quinasa I-kappa B/deficiencia , Quinasa I-kappa B/metabolismo , Inmunización , Recuento de Leucocitos , Ganglios Linfáticos/inmunología , Ratones , Ratones Transgénicos , Membrana Mucosa/metabolismo , Células Mieloides/inmunología , Células Mieloides/metabolismo , Infiltración Neutrófila/inmunología , Neutrófilos/metabolismo , Transducción de SeñalRESUMEN
AIMS/HYPOTHESIS: T helper type (Th) 17 cells have been shown to play important roles in mouse models of several autoimmune diseases that have been classified as Th1 diseases. In the NOD mouse, the relevance of Th1 and Th17 is controversial, because single-cytokine-deficient NOD mice develop diabetes similarly to wild-type NOD mice. METHODS: We studied the impact of IL-17/IFN-γ receptor double deficiency in NOD mice on the development of insulitis/diabetes compared with IL-17 single-deficient mice and wild-type mice by monitoring diabetes-related phenotypes. The lymphocyte phenotypes were determined by flow cytometric analysis. RESULTS: IL-17 single-deficient NOD mice showed delayed onset of diabetes and reduced severity of insulitis, but the cumulative incidence of longstanding diabetes in the IL-17-deficient mice was similar to that in wild-type mice. The IL-17/IFN-γ receptor double-deficient NOD mice showed an apparent decline in longstanding diabetes onset, but not in insulitis compared with that in the IL-17 single-deficient mice. We also found that double-deficient NOD mice had a severe lymphopenic phenotype and preferential increase in regulatory T cells among CD4(+) T cells compared with the IL-17 single-deficient mice and wild-type NOD mice. An adoptive transfer study with CD4(+)CD25(-) T cells from young non-diabetic IL-17 single-deficient NOD mice, but not those from older mice, showed significantly delayed disease onset in immune-deficient hosts compared with the corresponding wild-type mice. CONCLUSIONS/INTERPRETATION: These results indicate that IL-17/Th17 participates in the development of insulitis and that both IL-17 and IFN-γ signalling may synergistically contribute to the development of diabetes in NOD mice.
Asunto(s)
Diabetes Mellitus Tipo 1/genética , Interferón gamma/deficiencia , Interleucina-17/deficiencia , Traslado Adoptivo , Animales , Autoanticuerpos/genética , Autoanticuerpos/inmunología , Diabetes Mellitus Tipo 1/inmunología , Femenino , Citometría de Flujo , Interferón gamma/genética , Interleucina-17/genética , Masculino , Ratones , Ratones Endogámicos NOD , Ratones Mutantes , Ratones SCIDRESUMEN
Ligands for ErbB receptors, including epidermal growth factor (EGF) and neuregulin-1, have a neurotrophic activity on midbrain dopaminergic neurons and are implicated in the pathophysiology of schizophrenia. Although ErbB kinase inhibitors ameliorate behavioral deficits of the schizophrenia model that was established by hippocampal lesioning of rat pups, the antipsychotic action of ErbB kinase inhibitors and its general applicability to other models are not fully characterized. Using a different animal model, here, we examined whether and how ErbB kinase inhibitors ameliorate the behavioral endophenotypes relevant to schizophrenia. The animal model for schizophrenia was prepared by exposing neonatal rats to the cytokine EGF. Intraventricular infusion of the ErbB1 inhibitors ZD1839 and PD153035 in these animals ameliorated the deficits in startle response and prepulse inhibition in a dose-dependent manner. The deficits of latent inhibition of fear learning were also alleviated by ZD1839 with its limited effects on body weight gain or locomotor activity. ZD1839 infusion also decreased the busting activity of nigral dopamine (DA) neurons and reduced pallidal DA metabolism, a result that mimics the anti-dopaminergic profile of risperidone and haloperidol in this brain region. ErbB inhibitors appear to have anti-dopaminergic actions to alleviate some of the behavioral deficits common to animal models for schizophrenia.
Asunto(s)
Antipsicóticos/uso terapéutico , Neuronas Dopaminérgicas/efectos de los fármacos , Receptores ErbB/antagonistas & inhibidores , Quinazolinas/uso terapéutico , Esquizofrenia/tratamiento farmacológico , Animales , Química Encefálica/efectos de los fármacos , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Factor de Crecimiento Epidérmico/análisis , Factor de Crecimiento Epidérmico/farmacología , Femenino , Gefitinib , Masculino , Actividad Motora/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Reflejo de Sobresalto/efectos de los fármacos , Psicología del EsquizofrénicoRESUMEN
Calcineurin-inhibitor refractory bronchiolitis obliterans (BO) represents the leading cause of late graft failure after lung transplantation. T helper (Th)2 and Th17 lymphocytes have been associated with BO development. Taking advantage of a fully allogeneic trachea transplantation model in mice, we addressed the pathogenicity of Th cells in obliterative airway disease (OAD) occurring in cyclosporine A (CsA)-treated recipients. We found that CsA prevented CD8(+) T cell infiltration into the graft and downregulated the Th1 response but affected neither Th2 nor Th17 responses in vivo. In secondary mixed lymphocyte cultures, CsA dramatically decreased donor-specific IFN-γ production, enhanced IL-17 production and did not affect IL-13. As CD4(+) depletion efficiently prevented OAD in CsA-treated recipients, we further explored the role of Th2 and Th17 immunity in vivo. Although IL-4 and IL-17 deficient untreated mice developed an OAD comparable to wild-type recipients, a single cytokine deficiency afforded significant protection in CsA-treated recipients. In conclusion, CsA treatment unbalances T helper alloreactivity and favors Th2 and Th17 as coexisting pathways mediating chronic rejection of heterotopic tracheal allografts.
Asunto(s)
Bronquiolitis Obliterante/inducido químicamente , Ciclosporina/toxicidad , Rechazo de Injerto/inducido químicamente , Interleucina-17/fisiología , Trasplante de Pulmón/efectos adversos , Células Th2/inmunología , Tráquea/trasplante , Animales , Western Blotting , Bronquiolitis Obliterante/inmunología , Bronquiolitis Obliterante/patología , Citocinas/metabolismo , Citometría de Flujo , Rechazo de Injerto/inmunología , Rechazo de Injerto/patología , Técnicas para Inmunoenzimas , Inmunosupresores/toxicidad , Interferón gamma/fisiología , Interleucina-4/fisiología , Prueba de Cultivo Mixto de Linfocitos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tráquea/efectos de los fármacos , Tráquea/inmunología , Trasplante Heterotópico , Trasplante HomólogoRESUMEN
We determined whether a potential probiotic bacterium, Bifidobacterium bifidum OLB6378 (BB6378), exerts beneficial effects on the mucosal immune system in a mouse intestinal explant model. The addition of heat-inactivated BB6378 to intestinal explants prepared from embryonic day 18 BALB/c mice increased the expression of polymeric immunoglobulin receptor (pIgR) mRNA by two- to fivefold. These effects were observed on ileal and colonic explants but not on jejunal explants, suggesting that the BB6378-induced pIgR upregulation is site-specific within the mouse intestine. The upregulation of pIgR protein expression in colonic explants was also detected after 24 h of culture. The results of DNA microarray analysis of ileal and colonic samples indicated that BB6378 increased the gene expression of interleukin (IL)-1α and IL-1ß, and IL-1α content in colonic explants was significantly increased after 20 h of culture with BB6378. We then examined the involvement of endogenously induced IL-1α in pIgR mRNA upregulation by using IL-1α knockout (KO) mice. Contrary to our expectations, pIgR mRNA expression was equally upregulated by BB6378 in colonic explants from BALB/c and IL-1α KO mice. Conversely, we examined the involvement of Toll-like receptors in pIgR mRNA upregulation by using MyD88 KO mice. The upregulation of pIgR was completely suppressed in the explants derived from MyD88 KO mice. Taken together, we conclude that in a mouse intestinal explant model, the heat-inactivated potential probiotic BB6378 increases intestinal pIgR expression in a site-specific manner and that the upregulation of pIgR could be explained by a direct microbial effect on the epithelium via Toll-like receptors.
Asunto(s)
Bifidobacterium/inmunología , Mucosa Intestinal/efectos de los fármacos , Factor 88 de Diferenciación Mieloide/genética , Probióticos/farmacología , ARN Mensajero/genética , Receptores de Inmunoglobulina Polimérica/genética , Animales , Colon/efectos de los fármacos , Colon/inmunología , Colon/metabolismo , Embrión de Mamíferos , Calor , Íleon/efectos de los fármacos , Íleon/inmunología , Íleon/metabolismo , Inmunidad Mucosa/efectos de los fármacos , Interleucina-1alfa/genética , Interleucina-1alfa/inmunología , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Yeyuno/efectos de los fármacos , Yeyuno/inmunología , Yeyuno/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Factor 88 de Diferenciación Mieloide/deficiencia , Factor 88 de Diferenciación Mieloide/inmunología , Especificidad de Órganos , ARN Mensajero/inmunología , Receptores de Inmunoglobulina Polimérica/inmunología , Técnicas de Cultivo de Tejidos , Regulación hacia Arriba/inmunologíaRESUMEN
Interleukin-1 (IL-1) plays a crucial role in stress responses and its mRNA is induced in the brain by stress load; however, the precise role of IL-1 in higher brain functions and their abnormalities is largely unknown. Here, we report that IL-1 receptor antagonist (IL-1Ra) knockout (KO) mice, which lack IL-1Ra molecules that antagonize the IL-1 receptor, displayed anti-depression-like phenotypes in the tail suspension test (TST) and forced-swim test (FST) only at a young stage (8 weeks), whereas the phenotypes disappeared at later stages (20 and 32 weeks). These anti-depression-like phenotypes were reversed by administration of adrenergic receptor (AR) antagonists against the ARα(1), ARα(2), and ARß subtypes. Although the contents of 5-HT, norepinephrine (NE), and dopamine (DA), which are known to be associated with major symptoms of psychiatric disorders, were not significantly different in the hippocampus or cerebral cortex between IL-1Ra KO and their wild-type (WT) littermate mice, the mRNA expression level of the ARα(1A) subtype was significantly changed in the cerebral cortex. Interestingly, the change in expression of the ARα(1A) subtype was correlated with an age-dependent alteration in the TST and FST in IL-1Ra KO mice. Furthermore, mild immobilization stress loaded on C57BL/6J male mice caused similar anti-depression-like phenotypes in the TST and FST to those observed in mutant mice. These results suggest that sustained activation of IL-1 signaling induced by gene manipulation in mutant mice affects the expression of the ARα(1A) subtype and that modification of adrenergic signaling by the IL-1 system may ultimately cause significant psychiatric abnormalities such as depression, and this mutant mouse could be regarded as a model animal of depression that specifically appears in children and adolescents.
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Depresión/metabolismo , Proteína Antagonista del Receptor de Interleucina 1/metabolismo , Interleucina-1/metabolismo , Receptores Adrenérgicos alfa 1/metabolismo , Transducción de Señal/fisiología , Animales , Cromatografía Líquida de Alta Presión , Depresión/genética , Modelos Animales de Enfermedad , Proteína Antagonista del Receptor de Interleucina 1/genética , Interleucina-1/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores Adrenérgicos alfa 1/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: Inflammasome activation with the production of IL-1ß received substantial attention recently in inflammatory diseases. However, the role of inflammasome in the pathogenesis of asthma is not clear. Using an adjuvant-free model of allergic lung inflammation induced by ovalbumin (OVA), we investigated the role of NLRP3 inflammasome and related it to IL-1R1 signaling pathway. METHODS: Allergic lung inflammation induced by OVA was evaluated in vivo in mice deficient in NLRP3 inflammasome, IL-1R1, IL-1ß or IL-1α. Eosinophil recruitment, Th2 cytokine, and chemokine levels were determined in bronchoalveolar lavage fluid, lung homogenates, and mediastinal lymph node cells ex vivo. RESULTS: Allergic airway inflammation depends on NLRP3 inflammasome activation. Dendritic cell recruitment into lymph nodes, Th2 lymphocyte activation in the lung and secretion of Th2 cytokines and chemokines are reduced in the absence of NLRP3. Absence of NLRP3 and IL-1ß is associated with reduced expression of other proinflammatory cytokines such as IL-5, IL-13, IL-33, and thymic stromal lymphopoietin. Furthermore, the critical role of IL-1R1 signaling in allergic inflammation is confirmed in IL-1R1-, IL-1ß-, and IL-1α-deficient mice. CONCLUSION: NLRP3 inflammasome activation leading to IL-1 production is critical for the induction of a Th2 inflammatory allergic response.
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Asma/etiología , Proteínas Portadoras/inmunología , Inflamasomas/inmunología , Adyuvantes Inmunológicos , Aluminio , Animales , Asma/patología , Interleucina-1/biosíntesis , Interleucina-1beta , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR , Ovalbúmina , Neumonía , Receptores de Interleucina-1/metabolismo , Transducción de Señal/inmunología , Células Th2/inmunologíaRESUMEN
We have developed an animal model of learning and memory impairment associated with activation of microglia in the mouse brain. Injection of lipopolysaccharide into the CA1 region of the mouse hippocampus resulted in an increased production of inflammatory cytokines, such as interleukin-1ß. Immunostaining for interleukin-1ß revealed an increase in the signal at 6 hr after lipopolysaccharide injection. Immunopositive cells for interleukin-1ß were colocalized with those immunopositive for CD11b. When subacute lipopolysaccharide treatment (20 µg/2 µl/injection, bilaterally for 5 consecutive days) was performed, long-term activation of microglia and learning and memory deficits as evaluated using a step-through passive avoidance test were observed in the wild-type mice. Gene expression of the N-methyl-D-aspartate receptor NR1 and NR2A subunits was also decreased by the lipopolysaccharide treatment. In contrast, activation of microglia and the associated behavioral deficits were not observed in mice lacking interleukin-1α and -1ß following the subacute lipopolysaccharide treatment, together with little change in the gene expression of NR1 and NR2A subunits. However, the subacute lipopolysaccharide treatment produced almost similar changes in those parameters in the tumor necrosis factor-α knockout mice as in the wild-type animals. The injection of interleukin-1ß neutralizing antibody with lipopolysaccharide for 5 consecutive days resulted in the improvement of lipopolysaccharide-induced learning and memory deficits. These findings suggest that the expression of interleukin-1 plays an important role in lipopolysaccharide-induced activation of microglia and the associated functional deficits in learning and memory.
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Interleucina-1/metabolismo , Discapacidades para el Aprendizaje/metabolismo , Lipopolisacáridos/toxicidad , Trastornos de la Memoria/metabolismo , Microglía/metabolismo , Animales , Modelos Animales de Enfermedad , Expresión Génica , Hipocampo/efectos de los fármacos , Hipocampo/inmunología , Hipocampo/metabolismo , Inmunohistoquímica , Inyecciones Intraventriculares , Interleucina-1/inmunología , Discapacidades para el Aprendizaje/inmunología , Lipopolisacáridos/administración & dosificación , Lipopolisacáridos/inmunología , Masculino , Trastornos de la Memoria/inmunología , Ratones , Ratones Noqueados , Microglía/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
AIMS/HYPOTHESIS: Proinflammatory cytokines, including IL-1, exert pleiotropic effects on the neuro-immuno-endocrine system. Previously, we showed that mice with knockout of the gene encoding IL-1 receptor antagonist (Il1ra (-/-), also known as Il1rn (-/-)) have a lean phenotype. The present study was designed to analyse the mechanisms leading to this lean phenotype. METHODS: Il1ra (-/-) mice were fed a high-fat diet following weaning. Energy expenditure, body temperature, heart rate, blood parameters, urinary catecholamines and adipose tissue were analysed. RESULTS: Il1ra (-/-) mice exhibited resistance to obesity induced by a high-fat diet; this resistance was associated with increased energy expenditure and a decreased respiratory quotient, indicating that the ratio of fat:carbohydrate metabolism in Il1ra (-/-) mice is greater than in controls. Activity level in Il1ra (-/-) mice was significantly decreased and body temperature was significantly increased, compared with wild-type (WT) mice. Inguinal white adipose tissues in Il1ra (-/-) mice express increased levels of Ucp1 and mitochondrial respiratory chain genes compared with WT mice. Histological analysis of adipose tissue in Il1ra (-/-) mice revealed that brown adipose tissue is hyperactive and inguinal white adipose tissue contains smaller cells, which exhibit the distinctive multilocular appearance of brown adipocytes. Urinary epinephrine and norepinephrine excretion in Il1ra (-/-) mice was significantly increased compared with WT mice, suggesting that Il1ra (-/-) mice have increased sympathetic tone. Consistent with this, heart rate in Il1ra (-/-) mice was also significantly increased. CONCLUSIONS/INTERPRETATION: Our results show that Il1ra (-/-) mice have increased energy expenditure, fat:carbohydrate oxidation ratio, body temperature, heart rate and catecholamine production. All of these observations are consistent with an enhanced sympathetic tone.
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Carbohidratos de la Dieta/metabolismo , Grasas de la Dieta/metabolismo , Proteína Antagonista del Receptor de Interleucina 1/deficiencia , Tejido Adiposo Pardo/patología , Tejido Adiposo Pardo/fisiopatología , Animales , Metabolismo Energético , Epinefrina/orina , Inflamación/patología , Inflamación/fisiopatología , Proteína Antagonista del Receptor de Interleucina 1/genética , Proteína Antagonista del Receptor de Interleucina 1/metabolismo , Ratones , Ratones Noqueados , Norepinefrina/orina , Sistema Nervioso Simpático/fisiopatologíaRESUMEN
BACKGROUND: Few adequate murine models exist for metal allergies, it being especially difficult to induce Ni allergy in mice. OBJECTIVE: We examined the effect of lipopolysaccharide (LPS) on allergies to Ni and other metals in mice. METHODS: Ten days after sensitization with a metal salt and LPS, the ears were challenged with the same metal salt. RESULTS: LPS+NiCl(2) (1 mM) was effective at sensitizing mice to Ni, LPS being effective at very low concentrations whether injected intradermally or intraperitoneally. The ear-swelling response to Ni was more severe and more rapid in C57BL/6 mice than in BALB/c mice. In mast-cell-deficient mice, TNF-alpha-deficient mice, and interestingly even in nude (T cell deficient) mice, NiCl(2)+LPS induced a Ni allergy similar in degree to that in the respective control mice, but it induced Ni allergy only weakly in TLR4-mutant mice, macrophage-depleted mice, and IL-1-deficient mice. The activity of the histamine-forming enzyme histidine decarboxylase (HDC) in the ears increased in parallel with ear swelling, and HDC-deficient mice were resistant to ear swelling. Challenge with NiCl(2)+LPS augmented ear swelling (vs. NiCl(2) alone). LPS induced effective sensitization to other metals (Cr, Co, Pd, or Ag). CONCLUSIONS: These results indicate that in mice, LPS is a very important inducer of metal allergies, and potently promotes them (dependent on both innate immunity and HDC induction in cells other than mast cells). We discussed the idea that the bacterial environment is important for the establishment of metal allergies and for their provocation, and that the current thinking (including the contribution of T cells) should be reappraised in future studies.
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Dermatitis Alérgica por Contacto/etiología , Histidina Descarboxilasa/fisiología , Lipopolisacáridos/inmunología , Metales/inmunología , Adyuvantes Inmunológicos , Animales , Dermatitis Alérgica por Contacto/enzimología , Dermatitis Alérgica por Contacto/inmunología , Modelos Animales de Enfermedad , Peróxido de Hidrógeno/inmunología , Inmunidad Innata , Mastocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Níquel/inmunología , Especificidad de la EspecieRESUMEN
Cytokines may be crucially involved in the pathogenesis of inflammatory bowel diseases (IBD), but it remains controversial whether interferon (IFN)-gamma, a typical proinflammatory cytokine, is an essential mediator to cause the disorders. In the present study, IFN-gamma(-/-) and wild-type (WT) C57BL/6 mice were fed 2.5% dextran sodium sulphate (DSS) in drinking water for 7 days, in order to investigate DSS-induced intestinal inflammation. The DSS-treated WT mice exhibited a robust production of IFN-gamma in the gut, a remarkable loss of body weight, as well as high rate of mortality (60%). In striking contrast, IFN-gamma deficient mice did not develop DSS-induced colitis, as indicated by the maintenance of body weight and survival rate of 100%. Severe intestinal inflammation was demonstrated exclusively in WT animals in terms of the shortening of the bowel as well as the elevation of the disease activity index, myeloperoxidase (MPO) activity and serum haptoglobin level. Histological study of DSS-treated WT intestine revealed disruption of mucosal epithelium and massive infiltration of inflammatory cells, while the organ from IFN-gamma(-/-) mice remained virtually normal in appearance. Enzyme-linked immunosorbent assay (ELISA) analyses indicated abundant production of three chemokines, i.e. monokine induced by interferon-gamma (MIG), interferon-inducible protein 10 (IP-10) and monocyte chemoattractant protein-1 (MCP-1), in the DSS-irritated intestine of WT but not of IFN-gamma(-/-) mice. The present results demonstrate clearly that IFN-gamma plays indispensable roles in the initiation of DSS colitis, and some chemokines are produced in an IFN-gamma-dependent fashion.