RESUMEN
Mast cells (MCs) play critical roles in the establishment of allergic diseases. We recently demonstrated an unexpected, proinflammatory role for IL-10 in regulating MC responses. IL-10 enhanced MC activation and promoted IgE-dependent responses during food allergy. However, whether these effects extend to IgE-independent stimuli is not clear. In this article, we demonstrate that IL-10 plays a critical role in driving IL-33-mediated MC responses. IL-10 stimulation enhanced MC expansion and degranulation, ST2 expression, IL-13 production, and phospho-relA upregulation in IL-33-treated cells while suppressing TNF-α. These effects were partly dependent on endogenous IL-10 and further amplified in MCs coactivated with both IL-33 and IgE/Ag. IL-10's divergent effects also extended in vivo. In a MC-dependent model of IL-33-induced neutrophilia, IL-10 treatment enhanced MC responsiveness, leading to suppression of neutrophils and decreased TNF-α. In contrast, during IL-33-induced type 2 inflammation, IL-10 priming exacerbated MC activity, resulting in MC recruitment to various tissues, enhanced ST2 expression, induction of hypothermia, recruitment of eosinophils, and increased MCPT-1 and IL-13 levels. Our data elucidate an important role for IL-10 as an augmenter of IL-33-mediated MC responses, with implications during both allergic diseases and other MC-dependent disorders. IL-10 induction is routinely used as a prognostic marker of disease improvement. Our data suggest instead that IL-10 can enhance ST2 responsiveness in IL-33-activated MCs, with the potential to both aggravate or suppress disease severity depending on the inflammatory context.
Asunto(s)
Hipersensibilidad a los Alimentos , Mastocitos , Humanos , Mastocitos/metabolismo , Interleucina-10/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Inmunoglobulina E/metabolismo , Interleucina-33/metabolismo , Interleucina-13/metabolismo , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Inflamación/metabolismo , Degranulación de la CélulaAsunto(s)
Modelos Animales de Enfermedad , Hipersensibilidad a los Alimentos , Proteína 1 Similar al Receptor de Interleucina-1 , Interleucina-4 , Mastocitos , Animales , Mastocitos/inmunología , Mastocitos/metabolismo , Ratones , Hipersensibilidad a los Alimentos/inmunología , Interleucina-4/metabolismo , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Transducción de SeñalRESUMEN
IL-10 is a key pleiotropic cytokine that can both promote and curb Th2-dependent allergic responses. In this study, we demonstrate a novel role for IL-10 in promoting mast cell expansion and the development of IgE-mediated food allergy. Oral OVA challenge in sensitized BALB/c mice resulted in a robust intestinal mast cell response accompanied by allergic diarrhea, mast cell activation, and a predominance of Th2 cytokines, including enhanced IL-10 expression. In contrast, the development of intestinal anaphylaxis, including diarrhea, mast cell activation, and Th2 cytokine production, was significantly attenuated in IL-10(-/-) mice compared with wild-type (WT) controls. IL-10 also directly promoted the expansion, survival, and activation of mast cells; increased FcεRI expression on mast cells; and enhanced the production of mast cell cytokines. IL-10(-/-) mast cells had reduced functional capacity, which could be restored by exogenous IL-10. Similarly, attenuated passive anaphylaxis in IL-10(-/-) mice could be restored by IL-10 administration. The adoptive transfer of WT mast cells restored allergic symptoms in IL-10(-/-) mice, suggesting that the attenuated phenotype observed in these animals is due to a deficiency in IL-10-responding mast cells. Lastly, transfer of WT CD4 T cells also restored allergic diarrhea and intestinal mast cell numbers in IL-10(-/-) mice, suggesting that the regulation of IL-10-mediated intestinal mast cell expansion is T cell dependent. Our observations demonstrate a critical role for IL-10 in driving mucosal mast cell expansion and activation, suggesting that, in its absence, mast cell function is impaired, leading to attenuated food allergy symptoms.
Asunto(s)
Hipersensibilidad a los Alimentos/inmunología , Inmunoglobulina E/inmunología , Interleucina-10/inmunología , Mastocitos/inmunología , Traslado Adoptivo , Animales , Citocinas/biosíntesis , Citocinas/inmunología , Diarrea , Interleucina-10/administración & dosificación , Interleucina-10/deficiencia , Intestinos/citología , Intestinos/inmunología , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Ovalbúmina/inmunología , Receptores de IgE/genética , Células Th2/inmunologíaRESUMEN
Asthma is an immune-mediated disease of the airways characterized by reversible airway obstruction, bronchial eosinophilic inflammation, and airway hyperresponsiveness (AHR). The immune dysregulation in asthma has been attributed to the involvement of diverse immune cells that contribute to the immunopathology of the disease. Natural killer (NK) cells play critical roles in host defense against viruses and various cancers. Accumulating evidence demonstrates additional important roles for these cells in T cell priming, dendritic cell maturation, and the development of inflammation, all of which have the potential to enhance or dampen allergic responses. The ability of NK cells to produce Th2-type cytokines and their pivotal role in combating respiratory infections which cause airway dysfunction in asthmatics further suggest that they may directly contribute to the immunopathogenesis of allergic airway disease. In this review, we examine emerging evidence and discuss the putative roles of NK cells in the sensitization, progression, and resolution of asthma.
Asunto(s)
Asma/inmunología , Células Asesinas Naturales/inmunología , Alérgenos/inmunología , Animales , Citocinas/inmunología , Humanos , Fenotipo , Linfocitos T/inmunologíaRESUMEN
OBJECTIVE: To review the use of sofosbuvir for the treatment of chronic hepatitis C virus (HCV). DATA SOURCES: Review and nonreview articles were identified through MEDLINE (1996-April 2014), citations of articles, and meeting abstracts using keywords, including NS5B polymerase inhibitor, GS-7977, sofosbuvir, direct-acting antiviral (DAA), and others. STUDY SELECTION AND DATA EXTRACTION: Phase 1, 2, and 3 studies describing dose-ranging potential, pharmacokinetics, efficacy, safety, and tolerability of sofosbuvir were identified. DATA SYNTHESIS: Sofosbuvir is an NS5B polymerase inhibitor that was approved for use by the Food and Drug Administration in December 2013 for the treatment of chronic HCV in combination with pegylated interferon (peg-IFN) and ribavirin (RBV) for genotype 1. Additionally, it has been evaluated with other oral DAAs, such as simeprevir and others in the pipeline. It is not recommended as monotherapy because of lower sustained virological response (SVR) rates in clinical studies. Most of the treatment regimens are 12 weeks in duration; however, certain populations require a longer duration. Sofosbuvir has activity against all 6 genotypes, although most clinical trials evaluated genotypes 1 to 3. Sofosbuvir has a favorable safety and tolerability profile, making it a recommended first-line agent for chronic HCV infection. CONCLUSION: In clinical trials, 12 weeks of sofosbuvir with concomitant peg-IFN and RBV therapy in treatment-naïve and experienced HCV genotype 1 patients resulted in SVR rates of >90%. An all-oral regimen of sofosbuvir and RBV is highly effective for genotype 2 and 3 patients. Sofosbuvir was found to be tolerable with minimal adverse effects (AEs), and no treatment discontinuations occurred secondary to drug related AEs..
RESUMEN
IgE-mediated mast cell (MC) activation is a critical component of allergic responses to oral Ags. Several T cell-derived cytokines have been shown to promote MC reactivity, and we recently demonstrated a critical role for the cytokine IL-10 in mediating MC responses during food allergy. In this study, we further validate the role of IL-10 using Ab-mediated IL-10 depletion. IL-10 neutralization significantly attenuated MC responses, leading to decreased MC accumulation and activation, as well as inhibition of MC-mediated symptoms such as allergic diarrhea. This was accompanied by decreased Th2 cytokine gene expression, attenuated systemic T cell responses, and fewer CD4 T cells, B cells, and MCs in the spleen. Our data further confirm the role of IL-10 in driving MC responses and suggest that IL-10-responsive MCs may constitute an important player in allergic responses.
Asunto(s)
Modelos Animales de Enfermedad , Hipersensibilidad a los Alimentos , Interleucina-10 , Mastocitos , Animales , Femenino , Ratones , Anticuerpos Neutralizantes/inmunología , Linfocitos B/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Hipersensibilidad a los Alimentos/inmunología , Inmunoglobulina E/inmunología , Interleucina-10/metabolismo , Mastocitos/inmunología , Mastocitos/metabolismo , Ratones Endogámicos BALB C , Bazo/inmunología , Bazo/citología , Células Th2/inmunología , MasculinoRESUMEN
Atopic dermatitis (AD) is a common allergic inflammatory skin disease caused by a combination of intense pruritus, scratching, and epicutaneous (e.c.) sensitization with allergens. To explore the roles of IL-21 and IL-21 receptor (IL-21R) in AD, we examined skin lesions from patients with AD and used a mouse model of allergic skin inflammation. IL-21 and IL-21R expression was upregulated in acute skin lesions of AD patients and in mouse skin subjected to tape stripping, a surrogate for scratching. The importance of this finding was highlighted by the fact that both Il21r-/- mice and WT mice treated with soluble IL-21R-IgG2aFc fusion protein failed to develop skin inflammation after e.c. sensitization of tape-stripped skin. Adoptively transferred OVA-specific WT CD4+ T cells accumulated poorly in draining LNs (DLNs) of e.c. sensitized Il21r-/- mice. This was likely caused by both DC-intrinsic and nonintrinsic effects, because trafficking of skin DCs to DLNs was defective in Il21r-/- mice and, to a lesser extent, in WT mice reconstituted with Il21r-/- BM. More insight into this defect was provided by the observation that skin DCs from tape-stripped WT mice, but not Il21r-/- mice, upregulated CCR7 and migrated toward CCR7 ligands. Treatment of epidermal and dermal cells with IL-21 activated MMP2, which has been implicated in trafficking of skin DCs. These results suggest an important role for IL-21R in the mobilization of skin DCs to DLNs and the subsequent allergic response to e.c. introduced antigen.
Asunto(s)
Dermatitis Atópica , Inmunización , Inflamación/inmunología , Receptores de Interleucina-21/inmunología , Piel , Traslado Adoptivo , Animales , Células Cultivadas , Dermatitis Atópica/inmunología , Dermatitis Atópica/patología , Modelos Animales de Enfermedad , Activación Enzimática , Femenino , Humanos , Inflamación/inducido químicamente , Inflamación/patología , Interleucinas/genética , Interleucinas/inmunología , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 2 de la Matriz/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Ovalbúmina/inmunología , Ovalbúmina/farmacología , Receptores CCR7/genética , Receptores CCR7/inmunología , Receptores de Interleucina-21/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Piel/inmunología , Piel/patología , Bazo/citología , Bazo/inmunologíaRESUMEN
Individuals with atopic dermatitis immunized with the small pox vaccine, vaccinia virus (VV), are susceptible to eczema vaccinatum (EV), a potentially fatal disseminated infection. Dysfunction of Forkhead box P3 (FoxP3)-positive regulatory T cells (Treg) has been implicated in the pathogenesis of atopic dermatitis. To test whether Treg deficiency predisposes to EV, we percutaneously VV infected FoxP3-deficient (FoxP3(KO)) mice, which completely lack FoxP3(+) Treg. These animals generated both fewer VV-specific CD8(+) effector T cells and IFN-gamma-producing CD8(+) T cells than controls, had higher viral loads, and exhibited abnormal Th2-polarized responses to the virus. To focus on the consequences of Treg deficiency confined to the skin, we generated mixed CCR4(KO) FoxP3(KO) bone marrow (CCR4/FoxP3) chimeras in which skin, but not other tissues or central lymphoid organs, lack Treg. Like FoxP3(KO) mice, the chimeras had impaired VV-specific effector T cell responses and higher viral loads. Skin cytokine expression was significantly altered in infected chimeras compared with controls. Levels of the antiviral cytokines, type I and II IFNs and IL-12, were reduced, whereas expression of the proinflammatory cytokines, IL-6, IL-10, TGF-beta, and IL-23, was increased. Importantly, infection of CCR4/FoxP3 chimeras by a noncutaneous route (i.p.) induced immune responses comparable to controls. Our findings implicate allergic skin inflammation resulting from local Treg deficiency in the pathogenesis of EV.
Asunto(s)
Dermatitis Atópica/inmunología , Erupción Variceliforme de Kaposi/inmunología , Linfocitos T Reguladores/inmunología , Virus Vaccinia/inmunología , Animales , Trasplante de Médula Ósea , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Línea Celular , Citometría de Flujo , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/inmunología , Genoma Viral/genética , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/inmunología , Interferón gamma/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Reacción en Cadena de la Polimerasa , Receptores CCR4/genética , Receptores CCR4/inmunología , Piel/inmunología , Piel/patología , Piel/virología , Bazo/inmunología , Bazo/metabolismo , Bazo/patología , Linfocitos T Reguladores/metabolismo , Quimera por Trasplante/sangre , Quimera por Trasplante/inmunología , Vaccinia/inmunología , Virus Vaccinia/genéticaRESUMEN
BACKGROUND: In atopic subjects food ingestion drives the production of IgE antibodies that can trigger hypersensitivity reactions. The IL-4 pathway plays a critical role in this response, and genetic polymorphisms in its components have been linked to allergy. OBJECTIVE: We sought to test whether an activating mutation in the IL-4 receptor (IL-4R) α chain enhances allergic responses to a food antigen. METHODS: F709 mice, in which the IL-4Rα immunoreceptor tyrosine-based inhibitory motif is inactivated, were gavage fed with ovalbumin (OVA). Reactions to OVA challenge and immune responses, including antibody production and T(H)2 responses, were assessed. RESULTS: F709 mice, but not wild-type control animals, sensitized by means of gavage with OVA and either cholera toxin or staphylococcal enterotoxin B, displayed mast cell activation and systemic anaphylaxis on enteral challenge. Anaphylaxis was elicited even in F709 mice enterally sensitized with OVA alone. Bone marrow chimera experiments established that the increased sensitivity conferred by the F709 genotype was mediated mostly by hematopoietic cells but that nonhematopoietic cells also contributed. F709 mice exhibited increased intestinal permeability to macromolecules. The F709 genotype conferred increased OVA-specific IgE but not IgG1 responses, local and systemic T(H)2 responses, and intestinal mast cell hyperplasia compared with wild-type mice. Anaphylaxis was abrogated in F709 mice lacking IgE or the high-affinity receptor for IgE (FcεRI). CONCLUSION: Augmented IL-4Rα signaling confers increased intestinal permeability and dramatically enhanced sensitivity to food allergens. Unlike anaphylaxis to injected antigens, which in rodents can be mediated by either IgE or IgG antibodies, the food-induced response in F709 mice is solely IgE dependent.
Asunto(s)
Anafilaxia/inmunología , Hipersensibilidad a los Alimentos/inmunología , Inmunoglobulina E/inmunología , Receptores de Interleucina-4/inmunología , Transducción de Señal/inmunología , Anafilaxia/etiología , Animales , Ensayo de Inmunoadsorción Enzimática , Hipersensibilidad a los Alimentos/etiología , Ratones , Ratones Endogámicos BALB C , Células Th2/inmunología , Regulación hacia ArribaRESUMEN
Studies performed using cultured cells indicate that IgE functions not only to trigger degranulation of mast cells following allergen exposure, but also to enhance their survival. Such an influence of IgE on mast cell homeostasis during allergic responses in vivo has not been established. In this study, we show that inhalation of Aspergillus fumigatus extract in mice induced a dramatic rise in IgE accompanied by an increase in airway mast cells. These had an activated phenotype with high levels of FcepsilonRI. Plasma mast cell protease-1 was also increased, indicating an elevated systemic mast cell load. In addition, enhanced levels of IL-5 and eosinophils were observed in the airway. Both mast cell expansion and activation were markedly attenuated in IgE(-/-) animals that are incapable of producing IgE in response to A. fumigatus. The recruitment of eosinophils to the airways was also reduced in IgE(-/-) mice. Analyses of potential cellular targets of IgE revealed that IgE Abs are not required for the induction of mast cell progenitors in response to allergen, but rather act by sustaining the survival of mature mast cells. Our results identify an important role for IgE Abs in promoting mast cell expansion during allergic responses in vivo.
Asunto(s)
Hipersensibilidad/inmunología , Inmunoglobulina E/inmunología , Mastocitos/inmunología , Neumonía/inmunología , Células Madre/inmunología , Traslado Adoptivo , Animales , Aspergillus fumigatus/inmunología , Quimiotaxis de Leucocito/inmunología , Quimasas/inmunología , Quimasas/metabolismo , Ensayo de Inmunoadsorción Enzimática , Eosinófilos/inmunología , Citometría de Flujo , Homeostasis/inmunología , Hipersensibilidad/metabolismo , Interleucina-5/inmunología , Interleucina-5/metabolismo , Mastocitos/metabolismo , Ratones , Neumonía/metabolismo , Reacción en Cadena de la Polimerasa , Receptores de IgE/inmunología , Receptores de IgE/metabolismo , Células Madre/metabolismoRESUMEN
BACKGROUND: Signaling by IL-4 and IL-13 through the IL-4 receptor alpha chain (IL-4Ralpha) plays a critical role in the pathology of allergic diseases. The IL-4Ralpha is endowed with an immunoreceptor tyrosine-based inhibitory motif (ITIM) centered on tyrosine 709 (Y709) in the cytoplasmic domain that binds a number of regulatory phosphatases. The function of the ITIM in the in vivo regulation of IL-4 receptor signaling remains unknown. OBJECTIVE: We sought to determine the in vivo function of the IL-4Ralpha ITIM by using mice in which the ITIM was inactivated by mutagenesis of the tyrosine Y709 residue into phenylalanine (F709). METHODS: F709 ITIM mutant mice were derived by means of knock-in mutagenesis. Activation of intracellular signaling cascades by IL-4 and IL-13 was assessed by means of intracellular staining of phosphorylated signaling intermediates and gene expression analysis. In vivo responses to allergic sensitization were assessed by using models of allergic airway inflammation. RESULTS: The F709 mutation increased signal transducer and activator of transcription 6 phosphorylation by IL-4 and, disproportionately, by IL-13. This was associated with exaggerated T(H)2 polarization, enhanced alternative macrophage activation by IL-13, augmented basal and antigen-induced IgE responses, and intensified allergen-induced eosinophilic airway inflammation and hyperreactivity. CONCLUSIONS: These results point to a physiologic negative regulatory role for the Y709 ITIM in signaling through IL-4Ralpha, especially by IL-13.
Asunto(s)
Secuencias de Aminoácidos/inmunología , Inflamación/inmunología , Receptores de Superficie Celular , Hipersensibilidad Respiratoria/inmunología , Tirosina/química , Secuencias de Aminoácidos/genética , Animales , Humanos , Inflamación/etiología , Interleucina-13/genética , Interleucina-13/metabolismo , Interleucina-4/metabolismo , Ratones , Mutación , Fosforilación , Receptores de Superficie Celular/química , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Hipersensibilidad Respiratoria/etiología , Factor de Transcripción STAT6/genética , Factor de Transcripción STAT6/metabolismo , Transducción de SeñalRESUMEN
BACKGROUND: Atopic dermatitis and allergic contact dermatitis are skin disorders triggered by epicutaneous sensitization with protein antigens and contact sensitization with haptens, respectively. Skin is colonized with bacteria, which are a source of Toll-like receptor (TLR) 2 ligands. OBJECTIVE: We sought to examine the role of TLR2 in murine models of atopic dermatitis and allergic contact dermatitis. METHODS: TLR2(-/-) mice and wild-type littermates were epicutaneously sensitized with ovalbumin (OVA) or contact sensitized with oxazolone (OX). Skin histology was assessed by means of hematoxylin and eosin staining and immunohistochemistry. Ear swelling was measured with a micrometer. Cytokine mRNA expression was examined by means of quantitative RT-PCR. Antibody levels and splenocyte secretion of cytokines in response to OVA stimulation were measured by means of ELISA. Dendritic cells were examined for their ability to polarize T-cell receptor/OVA transgenic naive T cells to T(H)1 and T(H)2. RESULTS: In response to OVA sensitization, TLR2(-/-) mice experienced skin infiltration with eosinophils and CD4(+) cells, as well as upregulation of T(H)2 cytokine mRNAs that was comparable with that seen in wild-type littermates. In contrast, epidermal thickening, IFN-gamma expression in the skin, IFN-gamma production by splenocytes, and IgG2a anti-OVA antibody levels were impaired in TLR2(-/-) mice. After OX ear challenge, contact sensitized TLR2(-/-) mice exhibited defective ear swelling with impaired cellular infiltration, decreased epidermal thickening and local IFN-gamma expression, and impaired OX-specific IgG2a responses. Dendritic cells from TLR2(-/-) mice induced significantly lower production of IFN-gamma but normal IL-4 and IL-13 production in naive T cells. CONCLUSIONS: These results indicate that TLR2 promotes the IFN-gamma response to cutaneously introduced antigens.
Asunto(s)
Dermatitis Atópica/inmunología , Células TH1/inmunología , Receptor Toll-Like 2/fisiología , Animales , Femenino , Regulación de la Expresión Génica , Inmunoglobulina G/sangre , Interferón gamma/biosíntesis , Ratones , Ratones Endogámicos C57BL , Ovalbúmina/inmunología , Oxazolona/inmunología , Piel/inmunología , Piel/patología , Receptor Toll-Like 2/genéticaRESUMEN
Ellagic acid, a naturally occurring phenol found in a variety of fruits and nuts has been shown to possess anti-inflammatory properties. However, the mechanism of action behind its anti-inflammatory action is unclear. Using human Jurkat T cells, our study examined the effects of ellagic acid (EA) on Ca2+ handling, in particular, store-operated Ca2+ entry (SOCE), a process critical to proper T cell function. We observed that the acute addition of EA-induced Ca2+ release with an EC50 of 63 µM. The Ca2+ release was significantly attenuated by Xestospongin C, a known inhibitor of the Inositol 1,4,5-trisphosphate receptor (IP3R) channel and was unaffected by the phospholipase C (PLC) inhibitor, U73122. Furthermore, chronic incubation of Jurkat T cells with EA not only decreased the ATP-induced Ca2+ release but also diminished the SOCE-mediated Ca2+ influx in a dose-dependent manner. This inhibition was confirmed by reduced Mn2+ entry rates in the EA-treated cells. The ATP-induced Ca2+ entry was also attenuated in EA-treated HEK293 cells transiently transfected with SOCE channel Orai1-myc and ER-sensor stromal interaction molecule (STIM1) (HEKSTIM/Orai). Moreover, EA treatment interfered with the Orai1 and STIM1 coupling by disrupting STIM1 puncta formation in the HEKSTIM/Orai cells. We observed that EA treatment reduced cytokine secretion and nuclear factor of activated T-cell transcriptional activity in stimulated T cells. Hence, by inhibiting SOCE mediated Ca2+ influx, EA decreased downstream activation of pro-inflammatory mediators. These results suggest a novel target for EA-mediated effects and provide insight into the mechanisms underlying EA-mediated anti-inflammatory effects.
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Antiinflamatorios/farmacología , Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Ácido Elágico/farmacología , Señalización del Calcio/inmunología , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Estrenos/farmacología , Células HEK293 , Humanos , Mediadores de Inflamación/metabolismo , Receptores de Inositol 1,4,5-Trifosfato/metabolismo , Células Jurkat , Compuestos Macrocíclicos/farmacología , Proteínas de Neoplasias/metabolismo , Proteína ORAI1/metabolismo , Oxazoles/farmacología , Pirrolidinonas/farmacología , Molécula de Interacción Estromal 1/metabolismoRESUMEN
The thiol isomerase, protein disulfide isomerase (PDI), plays important intracellular roles during protein folding, maintaining cellular function and viability. Recent studies suggest novel roles for extracellular cell surface PDI in enhancing cellular activation and promoting their function. Moreover, a number of food-derived substances have been shown to regulate cellular PDI activity and alter disease progression. We hypothesized that PDI may have similar roles during mast cell-mediated allergic responses and examined its effects on IgE-induced mast cell activity during cell culture and food allergy. Mast cells were activated via IgE and antigen and the effects of PDI inhibition on mast cell activation were assessed. The effects of PDI blockade in vivo were examined by treating mice with the irreversible PDI inhibitor, PACMA-31, in an ovalbumin-induced model of food allergy. The role of dietary PDI modulators was investigated using various dietary compounds including curcumin and quercetin-3-rutinoside (rutin). PDI expression was observed on resting mast cell surfaces, intracellularly, and in the intestines of allergic mice. Furthermore, enhanced secretion of extracellular PDI was observed on mast cell membranes during IgE and antigen activation. Insulin turbidimetric assays demonstrated that curcumin is a potent PDI inhibitor and pre-treatment of mast cells with curcumin or established PDI inhibitors such as bacitracin, rutin or PACMA-31, resulted in the suppression of IgE-mediated activation and the secretion of various cytokines. This was accompanied by decreased mast cell proliferation, FcεRI expression, and mast cell degranulation. Similarly, treatment of allergic BALB/c mice with PACMA-31 attenuated the development of food allergy resulting in decreased allergic diarrhea, mast cell activation, and fewer intestinal mast cells. The production of TH2-specific cytokines was also suppressed. Our observations suggest that PDI catalytic activity is essential in the regulation of mast cell activation, and that its blockade may benefit patients with allergic inflammation.
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Antialérgicos/farmacología , Inhibidores Enzimáticos/farmacología , Hipersensibilidad a los Alimentos/prevención & control , Inmunoglobulina E/metabolismo , Intestinos/efectos de los fármacos , Mastocitos/efectos de los fármacos , Proteína Disulfuro Isomerasas/antagonistas & inhibidores , Animales , Bacitracina/farmacología , Degranulación de la Célula/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Curcumina/farmacología , Citocinas/metabolismo , Diarrea/enzimología , Diarrea/inmunología , Diarrea/prevención & control , Modelos Animales de Enfermedad , Hipersensibilidad a los Alimentos/enzimología , Hipersensibilidad a los Alimentos/inmunología , Intestinos/enzimología , Intestinos/inmunología , Mastocitos/enzimología , Mastocitos/inmunología , Ratones Endogámicos BALB C , Ovalbúmina , Proteína Disulfuro Isomerasas/metabolismo , Rutina/farmacología , Transducción de SeñalRESUMEN
Histamine, signaling via the type 1 receptor (H1R), has been shown to suppress Th2 cytokine production by in vitro cultured T cells. We examined the role of H1R in allergic inflammation in vivo using a murine asthma model. Allergen-stimulated splenic T cells from sensitized H1R-/- mice exhibited enhanced Th2 cytokine production. Despite this Th2 bias, allergen-challenged H1R-/- mice exhibited diminished lung Th2 cytokine mRNA levels, airway inflammation, goblet cell metaplasia, and airway hyperresponsiveness (AHR). Restoration of pulmonary Th2 cytokines in H1R-/- mice by intranasal IL-4 or IL-13 restored inflammatory lung responses and AHR. Further investigation revealed that histamine acts as a T cell chemotactic factor and defective T cell trafficking was responsible for the absence of lung inflammation. Cultured T cells migrated in response to histamine in vitro, but this was ablated by blockade of H1R but not H2R. In vivo, allergen-specific WT but not H1R-/- CD4+ T cells were recruited to the lungs of naive recipients following inhaled allergen challenge. H1R-/- T cells failed to confer airway inflammation or AHR observed after transfer of WT T cells. Our data establish a role for histamine and H1R in promoting the migration of Th2 cells into sites of allergen exposure.
Asunto(s)
Histamina/inmunología , Pulmón/inmunología , Receptores Histamínicos H1/inmunología , Hipersensibilidad Respiratoria/inmunología , Administración Intranasal , Animales , Movimiento Celular , Citocinas/inmunología , Modelos Animales de Enfermedad , Interleucina-13/administración & dosificación , Interleucina-13/inmunología , Interleucina-4/administración & dosificación , Interleucina-4/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ovalbúmina/inmunología , Receptores Histamínicos H1/genética , Linfocitos T/inmunología , Células Th2/inmunologíaRESUMEN
Mast cells are highly versatile cells that perform a variety of functions depending on the immune trigger, context of activation, and cytokine stimulus. Antigen-mediated mast cell responses are regulated by transcriptional processes that result in the induction of numerous genes contributing to mast cell function. Recently, we also showed that exposure to dietary agents with known epigenetic actions such as curcumin can suppress mast cell-mediated food allergy, suggesting that mast cell responses in vivo may be epigenetically regulated. To further assess the effects of epigenetic modifications on mast cell function, we examined the behavior of bone marrow-derived mast cells (BMMCs) in response to trichostatin A (TSA) treatment, a well-studied histone deacetylase inhibitor. IgE-mediated BMMC activation resulted in enhanced expression and secretion of IL-4, IL-6, TNF-α, and IL-13. In contrast, pretreatment with TSA resulted in altered cytokine secretion. This was accompanied by decreased expression of FcεRI and mast cell degranulation. Interestingly, exposure to non-IgE stimuli such as IL-33, was also affected by TSA treatment. Furthermore, continuous TSA exposure contributed to mast cell apoptosis and a decrease in survival. Further examination revealed an increase in I-κBα and a decrease in phospho-relA levels in TSA-treated BMMCs, suggesting that TSA alters transcriptional processes, resulting in enhancement of I-κBα transcription and decreased NF-κB activation. Lastly, treatment of wild-type mice with TSA in a model of ovalbumin-induced food allergy resulted in a significant attenuation in the development of food allergy symptoms including decreases in allergic diarrhea and mast cell activation. These data therefore suggest that the epigenetic regulation of mast cell activation during immune responses may occur via altered histone acetylation, and that exposure to dietary substances may induce epigenetic modifications that modulate mast cell function.
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Hipersensibilidad a los Alimentos/inmunología , Histonas/metabolismo , Mastocitos/inmunología , Acetilación , Animales , Apoptosis , Células de la Médula Ósea/citología , Degranulación de la Célula , Supervivencia Celular , Células Cultivadas , Citocinas/metabolismo , Modelos Animales de Enfermedad , Epigénesis Genética , Hipersensibilidad a los Alimentos/genética , Regulación de la Expresión Génica , Inhibidores de Histona Desacetilasas/metabolismo , Humanos , Ácidos Hidroxámicos/metabolismo , Inmunoglobulina E/metabolismo , Ratones , FN-kappa B/metabolismoRESUMEN
Histamine and its receptors (H1R-H4R) play a crucial and significant role in the development of various allergic diseases. Mast cells are multifunctional bone marrow-derived tissue-dwelling cells that are the major producer of histamine in the body. H1R are expressed in many cells, including mast cells, and are involved in Type 1 hypersensitivity reactions. H2R are involved in Th1 lymphocyte cytokine production. H3R are mainly involved in blood-brain barrier function. H4R are highly expressed on mast cells where their stimulation exacerbates histamine and cytokine generation. Both H1R and H4R have important roles in the progression and modulation of histamine-mediated allergic diseases. Antihistamines that target H1R alone are not entirely effective in the treatment of acute pruritus, atopic dermatitis, allergic asthma, and other allergic diseases. However, antagonists that target H4R have shown promising effects in preclinical and clinical studies in the treatment of several allergic diseases. In the present review, we examine the accumulating evidence suggesting novel therapeutic approaches that explore both H1R and H4R as therapeutic targets for histamine-mediated allergic diseases.
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Histamina/metabolismo , Hipersensibilidad/metabolismo , Inflamación/metabolismo , Mastocitos/inmunología , Receptores Histamínicos/metabolismo , Animales , Barrera Hematoencefálica , Citocinas/metabolismo , Antagonistas de los Receptores Histamínicos/uso terapéutico , Antagonistas de los Receptores Histamínicos H1/uso terapéutico , Humanos , Hipersensibilidad/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Terapia Molecular Dirigida , Receptores Histamínicos H4/antagonistas & inhibidores , Células TH1/inmunologíaRESUMEN
Objective. To implement a learner-led, discussion-based course aimed at exposing second-year pharmacy learners to the study of emerging infectious diseases from a global health perspective and to assess the role and importance of pharmacists in the management of disease outbreaks. Design. Learners examined literature pertinent to an emerging infectious disease in a 3-credit, discussion-based course and participated in peer discussion led by a designated learner. Instructional materials included journal articles, audio-visual presentations, documentaries, book chapters, movies, newspaper/magazine articles, and other materials. Learning outcomes were measured based on the ability of learners to perform critical thinking and analysis, communicate with their peers, and participate in class discussions. Assessment. The course was offered to 2 consecutive cohorts consisting of 14 and 16 learners, respectively. Overall, every learner in the first cohort achieved a final grade of A for the course. In the second cohort, the overall grade distribution consisted of grades of A, B, and C for the course. Learner evaluations indicated that the active-learning, discussion-based environment significantly enhanced interest in the topic and overall performance in the course. Conclusion. The elective course on emerging infectious diseases provided in-depth exposure to disease topics normally not encountered in the pharmacy curriculum. Learners found the material and format valuable, and the course enhanced their appreciation of infectious diseases, research methodology, critical thinking and analysis, and their roles as pharmacists.
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Enfermedades Transmisibles Emergentes , Curriculum , Educación en Farmacia/métodos , Aprendizaje Basado en Problemas/métodos , Estudiantes de Farmacia , Enfermedades Transmisibles Emergentes/diagnóstico , Enfermedades Transmisibles Emergentes/terapia , Curriculum/tendencias , Educación en Farmacia/tendencias , Humanos , Aprendizaje Basado en Problemas/tendenciasRESUMEN
IgE antibodies and mast cells play critical roles in the establishment of allergic responses to food antigens. Curcumin, the active ingredient of the curry spice turmeric, has anti-inflammatory properties, and thus may have the capacity to regulate Th2 cells and mucosal mast cell function during allergic responses. We assessed whether curcumin ingestion during oral allergen exposure can modulate the development of food allergy using a murine model of ovalbumin (OVA)-induced intestinal anaphylaxis. Herein, we demonstrate that frequent ingestion of curcumin during oral OVA exposure inhibits the development of mastocytosis and intestinal anaphylaxis in OVA-challenged allergic mice. Intragastric (i.g.) exposure to OVA in sensitized BALB/c mice induced a robust IgE-mediated response accompanied by enhanced OVA-IgE levels, intestinal mastocytosis, elevated serum mMCP-1, and acute diarrhea. In contrast, mice exposed to oral curcumin throughout the experimental regimen appeared to be normal and did not exhibit intense allergic diarrhea or a significant enhancement of OVA-IgE and intestinal mast cell expansion and activation. Furthermore, allergic diarrhea, mast cell activation and expansion, and Th2 responses were also suppressed in mice exposed to curcumin during the OVA-challenge phase alone, despite the presence of elevated levels of OVA-IgE, suggesting that curcumin may have a direct suppressive effect on intestinal mast cell activation and reverse food allergy symptoms in allergen-sensitized individuals. This was confirmed by observations that curcumin attenuated the expansion of both adoptively transferred bone marrow-derived mast cells (BMMCs), and inhibited their survival and activation during cell culture. Finally, the suppression of intestinal anaphylaxis by curcumin was directly linked with the inhibition of NF-κB activation in curcumin-treated allergic mice, and curcumin inhibited the phosphorylation of the p65 subunit of NF-κB in BMMCs. In summary, our data demonstrates a protective role for curcumin during allergic responses to food antigens, suggesting that frequent ingestion of this spice may modulate the outcome of disease in susceptible individuals.
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Anafilaxia/tratamiento farmacológico , Curcumina/uso terapéutico , Hipersensibilidad a los Alimentos/tratamiento farmacológico , Mastocitosis/tratamiento farmacológico , Anafilaxia/inmunología , Anafilaxia/metabolismo , Animales , Curcumina/farmacología , Modelos Animales de Enfermedad , Hipersensibilidad a los Alimentos/inmunología , Hipersensibilidad a los Alimentos/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/efectos de los fármacos , Intestinos/inmunología , Mastocitos/efectos de los fármacos , Mastocitos/inmunología , Mastocitos/metabolismo , Mastocitosis/inmunología , Mastocitosis/metabolismo , Ratones , FN-kappa B/metabolismo , Ovalbúmina , Fosforilación/efectos de los fármacos , Transducción de Señal/efectos de los fármacosRESUMEN
Polymorphisms in the interleukin-4 receptor alpha chain (IL-4R alpha) have been linked to asthma incidence and severity, but a causal relationship has remained uncertain. In particular, a glutamine to arginine substitution at position 576 (Q576R) of IL-4R alpha has been associated with severe asthma, especially in African Americans. We show that mice carrying the Q576R polymorphism exhibited intense allergen-induced airway inflammation and remodeling. The Q576R polymorphism did not affect proximal signal transducer and activator of transcription (STAT) 6 activation, but synergized with STAT6 in a gene target- and tissue-specific manner to mediate heightened expression of a subset of IL-4- and IL-13-responsive genes involved in allergic inflammation. Our findings indicate that the Q576R polymorphism directly promotes asthma in carrier populations by selectively augmenting IL-4R alpha-dependent signaling.