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1.
Eur J Drug Metab Pharmacokinet ; 44(3): 379-387, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30411300

RESUMO

BACKGROUND AND OBJECTIVES: Mast cell-mediated allergic diseases are a significant global health problem. Nitric oxide (NO) produced by acute type 1 allergies greatly suppresses hepatic cytochrome P450 (CYP) metabolism. A recent in vitro study demonstrated that repeated FcεRI-mediated activation intrinsically modulates mast cell function. We investigated the effect of ovalbumin (OVA) challenges on CYP activity and NO production under real immune responses. METHODS: After repeated sensitization with OVA once a week, serum nitrate plus nitrite (NOx) and total plasma immunoglobulin E concentrations were measured using commercially available kits. Hepatic microsomal CYP-specific activities and protein expression were determined using typical substrates and by western blot, respectively. In the liver, the levels of inducible NO synthase (iNOS), F4/80, and c-kit mRNA were determined by real-time polymerase chain reaction. Hepatic total NOS activity was measured using a colorimetric assay kit. RESULTS: When mice received multiple OVA challenges, the 11th sensitization elevated NOx concentrations in serum and suppressed the activities of five major CYPs without altering protein expression levels. After the 7th, 11th, and 15th sensitizations, F4/80-positive Kupffer cell and hepatic c-kit-dependent mast cell mRNA levels were similar to those of the control. The 7th and 11th sensitizations increased hepatic iNOS mRNA expression to 15-fold and threefold above control levels, respectively, but did not enhance the total NOS activity in the liver. CONCLUSIONS: Multiple OVA challenges, unlike acute sensitization, greatly reduced serum NOx levels. The challenge-suppressed hepatic CYP metabolism was likely related to the increased serum NOx. Serum NOx may be an endogenous marker for CYP metabolism inhibition in type 1 allergic diseases.


Assuntos
Sistema Enzimático do Citocromo P-450/metabolismo , Hipersensibilidade/enzimologia , Fígado/efeitos dos fármacos , Óxido Nítrico/biossíntese , Ovalbumina/imunologia , Animais , Sistema Enzimático do Citocromo P-450/imunologia , Feminino , Hipersensibilidade/sangue , Hipersensibilidade/imunologia , Imunoglobulina E/sangue , Fígado/enzimologia , Fígado/imunologia , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , Camundongos Endogâmicos ICR , Óxido Nítrico/sangue , Óxido Nítrico/imunologia , Óxido Nítrico Sintase Tipo II/metabolismo , Ovalbumina/administração & dosagem , Reação em Cadeia da Polimerase em Tempo Real
2.
Mol Pain ; 14: 1744806918767508, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29592783

RESUMO

Background Intense nociceptive signaling arising from ongoing injury activates primary afferent nociceptive systems to generate peripheral sensitization. ERK1/2 phosphorylation in dorsal root ganglion can be used to visualize intracellular signal activity immediately after noxious stimulation. The aim of this study was to investigate spatiotemporal characteristics of ERK1/2 phosphorylation against tissue injury in the primary afferent neurons. Methods Plantar incisions were made in the hind paws of Sprague-Dawley rats (n =150). Levobupivacaine was injected into the plantar aspect of the paws and ankles, Mitogen-activated protein kinase kinase (MEK) inhibitor was injected into the paw, and carbenoxolone, dual inhibitor of the gap junction and pannexin channel, was intraperitoneally injected. Pain hypersensitivity was investigated by a behavioral study, while phosphorylated ERK1/2 was detected in dorsal root ganglion and hind paw using immunohistochemistry and Western blot. Results Phosphorylated ERK1/2 was induced in dorsal root ganglion (26.8 ± 2.9% at baseline, 65.6 ± 3.6% at 2 min, and 26.3 ± 3.4% at 2 h) after the incision. NF-200 positive A-fiber neurons and satellite glial cells were positive for phosphorylated ERK1/2. Injury-induced pain hypersensitivity was abolished by MEK inhibitor. Levobupivacaine treatment inhibited phosphorylated ERK1/2 induction, carbenoxolone treatment inhibited glial phosphorylated ERK1/2 at 2 min after the injury, and carbenoxolone inhibited pain hypersensitivity and neuronal phosphorylated ERK1/2 at 1 h after the injury. Conclusion ERK1/2 phosphorylation in A-fiber neurons and satellite glial cells immediately after injury contributes to the generation of pain hypersensitivity. Signal communication between neurons and satellite glial cells expands the duration of neuronal ERK1/2 phosphorylation and pain hypersensitivity at 1 h after tissue injury.


Assuntos
MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Extremidades/patologia , Gânglios Espinais/enzimologia , Gânglios Espinais/patologia , Neuroglia/enzimologia , Neurônios/enzimologia , Dor/enzimologia , Analgésicos/farmacologia , Animais , Bupivacaína/farmacologia , Bupivacaína/uso terapêutico , Ativação Enzimática , Extremidades/cirurgia , Hipersensibilidade/enzimologia , Hipersensibilidade/patologia , Masculino , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Dor/tratamento farmacológico , Dor/patologia , Inibidores de Proteínas Quinases/farmacologia , Ratos Sprague-Dawley
3.
Gastroenterology ; 154(1): 140-153.e17, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28912017

RESUMO

BACKGROUND & AIMS: Chronic gastrointestinal inflammation increases the risk of cancer by mechanisms that are not well understood. Indoleamine-2,3-dioxygenase 1 (IDO1) is a heme-binding enzyme that regulates the immune response via catabolization and regulation of tryptophan availability for immune cell uptake. IDO1 expression is increased during the transition from chronic inflammation to gastric metaplasia. We investigated whether IDO1 contributes to the inflammatory response that mediates loss of parietal cells leading to metaplasia. METHODS: Chronic gastric inflammation was induced in Ido1-/- and CB57BL/6 (control) mice by gavage with Helicobacter felis or overexpression of interferon gamma in gastric parietal cells. We also performed studies in Jh-/- mice, which are devoid of B cells. Gastric tissues were collected and analyzed by flow cytometry, immunostaining, and real-time quantitative polymerase chain reaction. Plasma samples were analyzed by enzyme-linked immunosorbent assay. Gastric tissues were obtained from 20 patients with gastric metaplasia and 20 patients without gastric metaplasia (controls) and analyzed by real-time quantitative polymerase chain reaction; gastric tissue arrays were analyzed by immunohistochemistry. We collected genetic information on gastric cancers from The Cancer Genome Atlas database. RESULTS: H felis gavage induced significantly lower levels of pseudopyloric metaplasia in Ido1-/- mice, which had lower frequencies of gastric B cells, than in control mice. Blood plasma from H felis-infected control mice had increased levels of autoantibodies against parietal cells, compared to uninfected control mice, but this increase was lower in Ido1-/- mice. Chronically inflamed stomachs of Ido1-/- mice had significantly lower frequencies of natural killer cells in contact with parietal cells, compared with stomachs of control mice. Jh-/- mice had lower levels of pseudopyloric metaplasia than control mice in response to H felis infection. Human gastric pre-neoplasia and carcinoma specimens had increased levels of IDO1 messenger RNA compared with control gastric tissues, and IDO1 protein colocalized with B cells. Co-clustering of IDO1 messenger RNA with B-cell markers was corroborated by The Cancer Genome Atlas database. CONCLUSIONS: IDO1 mediates gastric metaplasia by regulating the B-cell compartment. This process appears to be associated with type II hypersensitivity/autoimmunity. The role of autoimmunity in the progression of pseudopyloric metaplasia warrants further investigation.


Assuntos
Gastrite/etiologia , Hipersensibilidade/etiologia , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Lesões Pré-Cancerosas/enzimologia , Neoplasias Gástricas/etiologia , Animais , Linfócitos B/fisiologia , Gastrite/enzimologia , Gastrite/patologia , Humanos , Hipersensibilidade/enzimologia , Hipersensibilidade/patologia , Metaplasia , Camundongos , Camundongos Endogâmicos C57BL , Lesões Pré-Cancerosas/patologia , Neoplasias Gástricas/enzimologia , Neoplasias Gástricas/patologia
4.
Int J Mol Sci ; 18(6)2017 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-28545251

RESUMO

Allergies arise from aberrant T helper type 2 responses to allergens. Several respiratory allergens possess proteolytic activity, which has been recognized to act as an adjuvant for the development of a Th2 response. Allergen source-derived proteases can activate the protease-activated receptor-2, have specific effects on immune cells by cleaving cell membrane-bound regulatory molecules, and can disrupt tight junctions. The protease activity can induce a non-allergen-specific inflammatory response in the airways, which will set the stage for an allergen-specific Th2 response. In this review, we will discuss the evidence for the induction of oxidative stress as an underlying mechanism in Th2 sensitization to proteolytic allergens. We will discuss recent data linking the proteolytic activity of an allergen to its potential to induce oxidative stress and how this can facilitate allergic sensitization. Based on experimental data, we propose that a less proficient anti-oxidant response to allergen-induced oxidative stress contributes to the susceptibility to allergic sensitization. Besides the effect of oxidative stress on the immune response, we will also discuss how oxidative stress can increase the immunogenicity of an allergen by chemical modification.


Assuntos
Alérgenos/imunologia , Estresse Oxidativo/fisiologia , Animais , Hipersensibilidade/enzimologia , Hipersensibilidade/genética , Hipersensibilidade/imunologia , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/metabolismo
5.
Expert Opin Ther Pat ; 27(8): 919-928, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28425830

RESUMO

INTRODUCTION: Tryptase is one of the main serine-proteinases located in the secretory granules of mast cells, and is released through degranulation, which is involved in the pathogenesis of allergic inflammatory disease, cardiovascular diseases, lung fibrosis and tumor. Therefore, inhibitors targeting tryptase may represent a new direction for the treatment of allergic inflammatory disease and other diseases. Areas covered: In this article, we discussed the history and development of tryptase inhibitors and described a variety of tryptase inhibitors via their structures and biological importance in clinical studies and drug development for tryptase-related diseases. Expert opinion: Initial tryptase inhibitors based on indole structure as the hydrophobic substituent on a benzylamine-piperidine template have low specificity and poor bioavailability. Therefore, designing new and specific inhibitors targeting tryptase should be involved in future clinical studies. Modifications toward indoles with varying N-substitution, introducing an amide bond, and growing the chain length contribute to an increase in the specific selectivity and potency of tryptase inhibitors. Tryptase has become the research hotspot to explore many related diseases. Therefore, there has been growing appreciation for the potential importance of the tryptase inhibitors as a target for treating these diseases.


Assuntos
Desenho de Drogas , Inibidores de Serino Proteinase/farmacologia , Triptases/antagonistas & inibidores , Animais , Humanos , Hipersensibilidade/tratamento farmacológico , Hipersensibilidade/enzimologia , Inflamação/tratamento farmacológico , Inflamação/enzimologia , Mastócitos/enzimologia , Mastócitos/metabolismo , Patentes como Assunto , Inibidores de Serino Proteinase/administração & dosagem , Triptases/metabolismo
6.
Gut ; 66(10): 1767-1778, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28096305

RESUMO

OBJECTIVES: Proteases are key mediators of pain and altered enteric neuronal signalling, although the types and sources of these important intestinal mediators are unknown. We hypothesised that intestinal epithelium is a major source of trypsin-like activity in patients with IBS and this activity signals to primary afferent and enteric nerves and induces visceral hypersensitivity. DESIGN: Trypsin-like activity was determined in tissues from patients with IBS and in supernatants of Caco-2 cells stimulated or not. These supernatants were also applied to cultures of primary afferents. mRNA isoforms of trypsin (PRSS1, 2 and 3) were detected by reverse transcription-PCR, and trypsin-3 protein expression was studied by western blot analysis and immunohistochemistry. Electrophysiological recordings and Ca2+ imaging in response to trypsin-3 were performed in mouse primary afferent and in human submucosal neurons, respectively. Visceromotor response to colorectal distension was recorded in mice administered intracolonically with trypsin-3. RESULTS: We showed that stimulated intestinal epithelial cells released trypsin-like activity specifically from the basolateral side. This activity was able to activate sensory neurons. In colons of patients with IBS, increased trypsin-like activity was associated with the epithelium. We identified that trypsin-3 was the only form of trypsin upregulated in stimulated intestinal epithelial cells and in tissues from patients with IBS. Trypsin-3 was able to signal to human submucosal enteric neurons and mouse sensory neurons, and to induce visceral hypersensitivity in vivo, all by a protease-activated receptor-2-dependent mechanism. CONCLUSIONS: In IBS, the intestinal epithelium produces and releases the active protease trypsin-3, which is able to signal to enteric neurons and to induce visceral hypersensitivity.


Assuntos
Células Epiteliais/enzimologia , Mucosa Intestinal/enzimologia , Síndrome do Intestino Irritável/enzimologia , Síndrome do Intestino Irritável/genética , Tripsina/genética , Tripsina/metabolismo , Animais , Células CACO-2 , Estudos de Casos e Controles , Colo/enzimologia , Colo/inervação , Meios de Cultivo Condicionados/farmacologia , Dipeptídeos/farmacologia , Sistema Nervoso Entérico/citologia , Sistema Nervoso Entérico/diagnóstico por imagem , Sistema Nervoso Entérico/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Feminino , Gânglios Espinais/citologia , Humanos , Hipersensibilidade/enzimologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/patologia , Isoxazóis/farmacologia , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Microscopia Confocal , Neurônios Aferentes/efeitos dos fármacos , Neurônios Aferentes/fisiologia , Permeabilidade/efeitos dos fármacos , RNA Mensageiro/análise , Ratos , Receptor PAR-2/antagonistas & inibidores , Receptor PAR-2/metabolismo , Tripsina/farmacologia , Tripsinogênio/genética , Regulação para Cima
7.
Chin J Integr Med ; 23(8): 570-573, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27460493

RESUMO

The histamine receptor antagonists in the treatment of allergic disease have limitations. The treatments of Chinese herbs have some curative effects on allergic skin lesions. Present research indicates that the mitogen-activated protein kinase (MAPK) signaling pathway might be equally important in allergic reactions. It was found that the inhibition of MAPK signaling pathways might relieve allergy symptoms, and some herbs can inhibit the MAPK pathway, which yields anti-allergy effects. Chinese medicines (CMs) have immense potential in the development of treatments for allergic disease.


Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Hipersensibilidade/tratamento farmacológico , Hipersensibilidade/enzimologia , Sistema de Sinalização das MAP Quinases , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Animais , Medicamentos de Ervas Chinesas/farmacologia , Humanos , Hipersensibilidade/imunologia , Imunomodulação/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Resultado do Tratamento
8.
Allergy ; 72(3): 425-434, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27460687

RESUMO

BACKGROUND: Diagnosis and immunotherapy of allergy against mites is based on complex extracts from large-scale cultures. However, the analysis of their composition using specific antibodies is limited. By taking advantage of the prevailing enzymatic nature of mite allergens, we have developed a broad-spectrum biochemical method for the standardization of native mite products. METHODS: Microplate-based assays have been implemented for thirteen Dermatophagoides pteronyssinus enzymatic activities, associated with Der p 1, 3, 4, 6, 8, 9, 15 and 20 allergens. The dynamics of these activities along culture growth, and their profile in purified fractions (bodies and faeces) and international reference standards (WHO/IUIS, two CBER/FDA), have been characterized. The stability of enzymatic activities and major allergens under stress conditions (40°C) has been assessed in the presence/absence of specific protease inhibitors. RESULTS: The analysis of enzymatic activities revealed distinct profiles along culture growth and between fractions (bodies and faeces). Remarkable differences were found when comparing international reference standards, being consistent with their source material (purified bodies or whole cultures). After 72 h at 40°C, only trypsin and alpha-amylase maintained high activity. Notably, the prominent role of trypsins in the hydrolytic degradation of major allergens is demonstrated by the use of inhibitors. CONCLUSIONS: Our method offers a robust approach to assess the complexity of mite extracts and highlights the critical importance of source materials for the composition and stability of finished products. The implementation of this approach in industry-based quality control procedures would contribute to the standardization of allergenic extracts used for diagnosis and immunotherapy.


Assuntos
Alérgenos/imunologia , Alérgenos/metabolismo , Hipersensibilidade/enzimologia , Hipersensibilidade/imunologia , Técnicas Imunoenzimáticas/métodos , Técnicas Imunoenzimáticas/normas , Pyroglyphidae/enzimologia , Pyroglyphidae/imunologia , Animais , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/normas , Cisteína Endopeptidases/imunologia , Cisteína Endopeptidases/normas , Dermatophagoides pteronyssinus/enzimologia , Dermatophagoides pteronyssinus/imunologia , Ativação Enzimática , Estabilidade Enzimática , Humanos , Hipersensibilidade/diagnóstico , Controle de Qualidade , Padrões de Referência , Especificidade por Substrato
10.
Curr Allergy Asthma Rep ; 16(12): 85, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27878551

RESUMO

PURPOSE OF REVIEW: The purpose of this review is to summarize the evidence from recently published original studies investigating how glutathione S-transferase (GST) gene polymorphisms modify the impact of air pollution on asthma, allergic diseases, and lung function. RECENT FINDINGS: Current studies in epidemiological and controlled human experiments found evidence to suggest that GSTs modify the impact of air pollution exposure on respiratory diseases and allergies. Of the nine articles included in this review, all except one identified at least one significant interaction with at least one of glutathione S-transferase pi 1 (GSTP1), glutathione S-transferase mu 1 (GSTM1), or glutathione S-transferase theta 1 (GSTT1) genes and air pollution exposure. The findings of these studies, however, are markedly different. This difference can be partially explained by regional variation in the exposure levels and oxidative potential of different pollutants and by other interactions involving a number of unaccounted environment exposures and multiple genes. Although there is evidence of an interaction between GST genes and air pollution exposure for the risk of respiratory disease and allergies, results are not concordant. Further investigations are needed to explore the reasons behind the discordancy.


Assuntos
Poluição do Ar/efeitos adversos , Glutationa Transferase/genética , Hipersensibilidade/etiologia , Transtornos Respiratórios/etiologia , Exposição Ambiental , Predisposição Genética para Doença , Humanos , Hipersensibilidade/enzimologia , Hipersensibilidade/genética , Polimorfismo Genético , Transtornos Respiratórios/enzimologia , Transtornos Respiratórios/genética , Fatores de Risco
11.
Blood ; 128(25): 2909-2918, 2016 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-27789480

RESUMO

Recent studies have identified nonredundant roles for basophils in immune responses including allergy and protective immunity. It is well known that activated basophils release granule contents such as histamine and proteases as do mast cells. However, the functional significance of basophil-derived proteases remains poorly understood in contrast to those released from mast cells. For this study we generated a line of knockout (KO) mice deficient for mouse mast cell protease-11 (mMCP-11) that is preferentially expressed by basophils rather than mast cells. In spite of normal development of basophils, the mMCP-11-deficient mice showed amelioration of immunoglobulin E-mediated chronic allergic inflammation (IgE-CAI), with reduction of cutaneous swelling, microvascular permeability, and leukocyte infiltration in the skin lesion, when KO mice were compared with wild-type mice. Repeated administration of recombinant mMCP-11 in the skin induced infiltration of leukocytes, including basophils, in a tryptase activity-dependent manner. The transwell migration assay in vitro suggested that mMCP-11-mediated proteolytic products of serum protein promoted migration of basophils, eosinophils, and macrophages via 1 or more G protein-coupled receptors. Thus, basophil tryptase mMCP-11 is a crucial effector molecule for the induction of IgE-CAI. This is the first demonstration that the basophil-derived protease plays a significant role in vivo.


Assuntos
Basófilos/enzimologia , Hipersensibilidade/enzimologia , Hipersensibilidade/imunologia , Imunoglobulina E/imunologia , Inflamação/enzimologia , Inflamação/imunologia , Triptases/metabolismo , Animais , Permeabilidade Capilar , Movimento Celular , Doença Crônica , Hipersensibilidade/complicações , Hipersensibilidade/patologia , Inflamação/complicações , Inflamação/patologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteólise , Receptores Acoplados a Proteínas-G/metabolismo , Pele/irrigação sanguínea , Pele/patologia , Triptases/deficiência
12.
PLoS One ; 11(7): e0159310, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27442134

RESUMO

The catalytical isoforms p110γ and p110δ of phosphatidylinositide 3-kinase γ (PI3Kγ) and PI3Kδ play an important role in the pathogenesis of asthma. Two key elements in allergic asthma are increased levels of eosinophils and IgE. Dual pharmacological inhibition of p110γ and p110δ reduces asthma-associated eosinophilic lung infiltration and ameliorates disease symptoms, whereas the absence of enzymatic activity in p110γKOδD910A mice increases IgE and basal eosinophil counts. This suggests that long-term inhibition of p110γ and p110δ might exacerbate asthma. Here, we analysed mice genetically deficient for both catalytical subunits (p110γ/δ-/-) and determined basal IgE and eosinophil levels and the immune response to ovalbumin-induced asthma. Serum concentrations of IgE, IL-5 and eosinophil numbers were significantly increased in p110γ/δ-/- mice compared to single knock-out and wildtype mice. However, p110γ/δ-/- mice were protected against OVA-induced infiltration of eosinophils, neutrophils, T and B cells into lung tissue and bronchoalveolar space. Moreover, p110γ/δ-/- mice, but not single knock-out mice, showed a reduced bronchial hyperresponsiveness. We conclude that increased levels of eosinophils and IgE in p110γ/δ-/- mice do not abolish the protective effect of p110γ/δ-deficiency against OVA-induced allergic airway inflammation.


Assuntos
Classe I de Fosfatidilinositol 3-Quinases/deficiência , Eosinofilia/enzimologia , Eosinofilia/imunologia , Imunoglobulina E/biossíntese , Ovalbumina/imunologia , Pneumonia/enzimologia , Pneumonia/imunologia , Animais , Linfócitos B/imunologia , Hiper-Reatividade Brônquica/sangue , Hiper-Reatividade Brônquica/complicações , Hiper-Reatividade Brônquica/enzimologia , Hiper-Reatividade Brônquica/imunologia , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Contagem de Células , Classe I de Fosfatidilinositol 3-Quinases/metabolismo , Eosinofilia/sangue , Eosinofilia/complicações , Eosinófilos/imunologia , Células Caliciformes/patologia , Hipersensibilidade/sangue , Hipersensibilidade/complicações , Hipersensibilidade/enzimologia , Hipersensibilidade/imunologia , Interleucina-5/sangue , Pulmão/imunologia , Pulmão/patologia , Metaplasia , Camundongos Endogâmicos C57BL , Neutrófilos/imunologia , Pneumonia/sangue , Pneumonia/complicações , Linfócitos T/imunologia
13.
Adv Med Sci ; 61(2): 300-305, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27149557

RESUMO

PURPOSE: This study aimed to investigate early-life folate serum concentrations in children with food, inhalant or mixed type allergy. The influence of folate levels on the FoxP3 expression in Treg (regulatory T) cells in the studied children, taking into account the MTHFR (5,10-methylenetetrahydrofolate reductase) genotypes was also analyzed. MATERIAL AND METHODS: The study was performed in 83 allergic children (study group) and 49 healthy children (control group), aged 2-72 months. Medical history of each child was obtained and laboratory tests (serum folic acid concentrations and MTHFR C677T polymorphism) were carried out. The percentage of Treg cells was evaluated in almost a half of the examined subjects (48.5%). RESULTS: Significantly higher serum folate levels in the group of children with food allergy than in those with inhalant allergy was confirmed (P=0.037). In the study group the TT homozygotes were characterized by significantly lower folate concentrations than CC homozygotes (P=0.045). A negative correlation was demonstrated between the FoxP3 expression in CD4+CD25highFoxP3+ peripheral blood lymphocytes and serum folic acid concentrations. The correlation was more pronounced in the group of allergic children and it was statistically significant (r=-0.339, P<0.05). CONCLUSIONS: The results of the study indicate a possibility of some effects of folate status on Treg cells, thus suggesting their potential role in the development and course of allergy in children.


Assuntos
Ácido Fólico/sangue , Hipersensibilidade/genética , Hipersensibilidade/imunologia , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Polimorfismo de Nucleotídeo Único/genética , Linfócitos T Reguladores/imunologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Fatores de Transcrição Forkhead/metabolismo , Humanos , Hipersensibilidade/sangue , Hipersensibilidade/enzimologia , Lactente , Contagem de Linfócitos , Masculino
14.
J Allergy Clin Immunol ; 138(4): 1170-1182.e9, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-26948079

RESUMO

BACKGROUND: IL-10-producing regulatory B (B10) cells potently suppress allergic diseases, such as contact hypersensitivity (CHS). Splenic B10 cells share overlapping phenotypic markers with CD5+ B1 B cells, CD1dhiCD21+CD23- marginal zone (MZ) B cells, and CD1dhiCD21+CD23+ T2-MZ precursor B cells but do not exclusively belong to either subset. OBJECTIVE: In this study we investigated the signaling mechanisms and a novel phenotypic parameter of B10 cells. METHOD: We performed microarray analysis comparing IL-10+ and IL-10- B cells. B cell-specific phosphatase and tensin homolog (PTEN)-deficient mice, which exhibit aberrant activation of the phosphatidylinositol 3-kinase (PI3K)-Akt pathway in B cells, were examined. RESULTS: Microarray analysis revealed that the PI3K-Akt pathway is important for IL-10 production in B cells. PI3K-Akt pathway inhibitors reduced B10 cell numbers in vitro. B10 cell numbers were significantly increased in B cell-specific PTEN-deficient mice. The CHS response was significantly diminished in PTEN-deficient mice. Unexpectedly, splenic B10 cells in these mice were found within the B1 B-cell subset but not within the MZ B-cell subset. In wild-type mice not only MZ B10 cells but also B1-B10 cells were identified in the spleen. In addition, these 2 B10 cell subsets were predominantly found within the CD9+CD80+ B-cell fraction. CONCLUSION: A novel splenic B1 regulatory cell subset (B1-B10 cells) was identified. Our findings show that the PI3K-Akt pathway in B cells is critical for B10 cell development and CHS response and that CD9/CD80 coexpression is a novel phenotypic parameter for both MZ-B10 and B1-B10 cells.


Assuntos
Linfócitos B/imunologia , Hipersensibilidade/enzimologia , Fosfatidilinositol 3-Quinase/imunologia , Proteínas Proto-Oncogênicas c-akt/imunologia , Animais , Linfócitos B/classificação , Linfócitos B/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Humanos , Hipersensibilidade/imunologia , Interleucina-10/metabolismo , Subpopulações de Linfócitos/efeitos dos fármacos , Subpopulações de Linfócitos/imunologia , Camundongos , Análise em Microsséries , Transdução de Sinais/efeitos dos fármacos , Baço/citologia , Baço/imunologia
15.
Immunol Cell Biol ; 94(7): 701-8, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27001469

RESUMO

Eosinophils contribute to immune regulation and wound healing/fibrosis in various diseases, including asthma. Growing appreciation for the role of activin A in such processes led us to hypothesize that eosinophils are a source of this transforming growth factor-ß superfamily member. Tumor necrosis factor-α (TNF) induces activin A by other cell types and is often present at the site of allergic inflammation along with the eosinophil-activating common ß (ßc) chain-signaling cytokines (interleukin (IL)-5, IL-3, granulocyte-macrophages colony-stimulating factor (GM-CSF)). Previously, we established that the combination of TNF plus a ßc chain-signaling cytokine synergistically induces eosinophil synthesis of the remodeling enzyme matrix metalloproteinase-9. Therefore, eosinophils were stimulated ex vivo by these cytokines and in vivo through an allergen-induced airway inflammatory response. In contrast to IL-5+TNF or GM-CSF+TNF, the combination of IL-3+TNF synergistically induced activin A synthesis and release by human blood eosinophils. IL-3+TNF enhanced activin A mRNA stability, which required sustained signaling of pathways downstream of p38 and extracellular signal-regulated kinase mitogen-activated protein kinases. In vivo, following segmental airway allergen challenge of subjects with mild allergic asthma, activin A mRNA was upregulated in airway eosinophils compared with circulating eosinophils, and ex vivo, circulating eosinophils tended to release more activin A in response to IL-3+TNF. These data provide evidence that eosinophils release activin A and that this function is enhanced when eosinophils are present in an allergen-induced inflammatory environment. Moreover, these data provide the first evidence for posttranscriptional control of activin A mRNA. We propose that an environment rich in IL-3+TNF will lead to eosinophil-derived activin A, which has an important role in regulating inflammation and/or fibrosis.


Assuntos
Ativinas/metabolismo , Eosinófilos/metabolismo , Interleucina-3/farmacologia , Estabilidade de RNA , Fator de Necrose Tumoral alfa/farmacologia , Adulto , Ativação Enzimática/efeitos dos fármacos , Eosinófilos/efeitos dos fármacos , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Hipersensibilidade/enzimologia , Hipersensibilidade/imunologia , Hipersensibilidade/patologia , Subunidades beta de Inibinas/genética , Subunidades beta de Inibinas/metabolismo , Interleucina-5/farmacologia , Cinética , Masculino , Pessoa de Meia-Idade , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Estabilidade de RNA/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Adulto Jovem
16.
Adv Med Sci ; 61(1): 141-6, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26686860

RESUMO

PURPOSE: To evaluate the role of 12/15-lipoxygenase (LOX) in regulation of synthesis of selected eicosanoids in mice sensitized and challenged with Dermatophagoides pteronyssinus (Dp) allergen extract. MATERIALS AND METHODS: Both C57Bl and 12/15-LOX knockout mice were sensitized by 2 intraperitoneal injections and subsequently challenged by inhalation with Dp allergen extract. Sham sensitized and challenged mice were used as controls. Samples of bronchoalveolar lavage (BAL) were used for assessment of prostaglandin E2 (PGE2), cysteinyl leukotreienes (cysLT), lipoxin A4 (LXA4) and 15-hydroxyeicosatetraenoic acid (15-HETE) concentration using ELISA method. Whole lung samples were used for isolation of RNA and evaluation of selected genes involved in eicosanoid metabolism, including cyclooxygenase-2 (COX-2), 12/15-LOX, 5-LOX and 5-LOX activated protein (FLAP). RESULTS: Allergen-induced airway inflammation was associated with significant (9-fold, 95% CI 8.068-9.932-fold; p<0.05) up-regulation of 12/15-LOX in wild type but not in the 12/15-LOX knockout mice in which 12/15-LOX mRNA remained undetectable. Lack of 12/15-LOX was associated with significant attenuation of production of 15-HETE in response to allergen challenge. On the contrary, the greatest up-regulation of COX-2 after allergen exposure was demonstrated in the 12/15-LOX knockout mice (4.3-fold vs sham group) and was significantly greater than in the wild type counterparts (5.185-fold, 95% CI 4.723-6.309-fold; p<0.05 vs wild type mice). Also, allergen challenged 12/15-LOX knockout mice were characterized by greater production of PGE2 and cysLT. CONCLUSION: The 12/15-LOX plays an important role in the metabolism of eicosanoids in response to allergen-induced airway inflammation.


Assuntos
Araquidonato 12-Lipoxigenase/metabolismo , Araquidonato 15-Lipoxigenase/metabolismo , Eicosanoides/biossíntese , Hipersensibilidade/complicações , Hipersensibilidade/enzimologia , Pneumonia/complicações , Pneumonia/enzimologia , Animais , Antígenos de Dermatophagoides , Vias Biossintéticas/genética , Líquido da Lavagem Broncoalveolar , Feminino , Regulação Enzimológica da Expressão Gênica , Imunoglobulina E/biossíntese , Pulmão/enzimologia , Pulmão/patologia , Camundongos Endogâmicos C57BL , Camundongos Knockout
17.
Science ; 349(6252): 1106-10, 2015 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-26339029

RESUMO

Growing up on a dairy farm protects children from allergy, hay fever, and asthma. A mechanism linking exposure to this endotoxin (bacterial lipopolysaccharide)-rich environment with protection has remained elusive. Here we show that chronic exposure to low-dose endotoxin or farm dust protects mice from developing house dust mite (HDM)-induced asthma. Endotoxin reduced epithelial cell cytokines that activate dendritic cells (DCs), thus suppressing type 2 immunity to HDMs. Loss of the ubiquitin-modifying enzyme A20 in lung epithelium abolished the protective effect. A single-nucleotide polymorphism in the gene encoding A20 was associated with allergy and asthma risk in children growing up on farms. Thus, the farming environment protects from allergy by modifying the communication between barrier epithelial cells and DCs through A20 induction.


Assuntos
Proteínas de Ligação a DNA/biossíntese , Poeira/imunologia , Hipersensibilidade/prevenção & controle , Peptídeos e Proteínas de Sinalização Intracelular/biossíntese , Lipopolissacarídeos/imunologia , Pulmão/enzimologia , Proteínas Nucleares/biossíntese , Pyroglyphidae/imunologia , Mucosa Respiratória/enzimologia , Animais , Asma/imunologia , Asma/prevenção & controle , Células Cultivadas , Criança , Indústria de Laticínios , Células Dendríticas/imunologia , Feminino , Humanos , Hipótese da Higiene , Hipersensibilidade/enzimologia , Hipersensibilidade/imunologia , Exposição por Inalação , Pulmão/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Mucosa Respiratória/imunologia , Proteína 3 Induzida por Fator de Necrose Tumoral alfa
18.
PLoS One ; 10(6): e0127032, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26039697

RESUMO

Heparanase is an endo-ß-glucuronidase that specifically cleaves heparan sulfate proteoglycans in the extracellular matrix. Expression of this enzyme is increased in several pathological conditions including inflammation. We have investigated the role of heparanase in pulmonary inflammation in the context of allergic and non-allergic pulmonary cell recruitment using heparanase knockout (Hpa-/-) mice as a model. Following local delivery of LPS or zymosan, no significant difference was found in the recruitment of neutrophils to the lung between Hpa-/- and wild type (WT) control. Similarly neutrophil recruitment was not inhibited in WT mice treated with a heparanase inhibitor. However, in allergic inflammatory models, Hpa-/- mice displayed a significantly reduced eosinophil (but not neutrophil) recruitment to the airways and this was also associated with a reduction in allergen-induced bronchial hyperresponsiveness, indicating that heparanase expression is associated with allergic reactions. This was further demonstrated by pharmacological treatment with a heparanase inhibitor in the WT allergic mice. Examination of lung specimens from patients with different severity of chronic obstructive pulmonary disease (COPD) found increased heparanase expression. Thus, it is established that heparanase contributes to allergen-induced eosinophil recruitment to the lung and could provide a novel therapeutic target for the development of anti-inflammatory drugs for the treatment of asthma and other allergic diseases.


Assuntos
Glucuronidase/metabolismo , Hipersensibilidade/enzimologia , Hipersensibilidade/patologia , Pulmão/enzimologia , Pulmão/patologia , Animais , Inibidores Enzimáticos/farmacologia , Feminino , Deleção de Genes , Glucuronidase/antagonistas & inibidores , Humanos , Imunização , Inflamação/patologia , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Masculino , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Ovalbumina , Doença Pulmonar Obstrutiva Crônica/enzimologia , Doença Pulmonar Obstrutiva Crônica/patologia , Testes de Função Respiratória
19.
PLoS One ; 10(6): e0129829, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26068872

RESUMO

Allergic diseases such as atopic dermatitis, rhinitis, asthma, and anaphylaxis are attractive research areas. Tyrosol (2-(4-hydroxyphenyl)ethanol) is a polyphenolic compound with diverse biological activities. In this study, we investigated whether tyrosol has anti-allergic inflammatory effects. Ovalbumin-induced active systemic anaphylaxis and immunoglobulin E-mediated passive cutaneous anaphylaxis models were used for the immediate-type allergic responses. Oral administration of tyrosol reduced the allergic symptoms of hypothermia and pigmentation in both animal models. Mast cells that secrete allergic mediators are key regulators on allergic inflammation. Tyrosol dose-dependently decreased mast cell degranulation and expression of inflammatory cytokines. Intracellular calcium levels and activation of inhibitor of κB kinase (IKK) regulate cytokine expression and degranulation. Tyrosol blocked calcium influx and phosphorylation of the IKK complex. To define the molecular target for tyrosol, various signaling proteins involved in mast cell activation such as Lyn, Syk, phosphoinositide 3-kinase (PI3K), and Akt were examined. Our results showed that PI3K could be a molecular target for tyrosol in mast cells. Taken together, these findings indicated that tyrosol has anti-allergic inflammatory effects by inhibiting the degranulation of mast cells and expression of inflammatory cytokines; these effects are mediated via PI3K. Therefore, we expect tyrosol become a potential therapeutic candidate for allergic inflammatory disorders.


Assuntos
Antioxidantes/farmacologia , Hipersensibilidade/prevenção & controle , Inflamação/prevenção & controle , Mastócitos/imunologia , Anafilaxia Cutânea Passiva/efeitos dos fármacos , Álcool Feniletílico/análogos & derivados , Animais , Antialérgicos/farmacologia , Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Western Blotting , Degranulação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Ativação Enzimática/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Hipersensibilidade/enzimologia , Hipersensibilidade/imunologia , Inflamação/enzimologia , Inflamação/imunologia , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/enzimologia , Camundongos , Anafilaxia Cutânea Passiva/imunologia , Álcool Feniletílico/farmacologia , Fosforilação/efeitos dos fármacos , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
20.
Drug Metab Dispos ; 43(8): 1169-80, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25994032

RESUMO

Members of the cytochrome P450 CYP2J subfamily are expressed in multiple tissues in mice and humans. These enzymes are active in the metabolism of fatty acids to generate bioactive compounds. Herein we report new methods and results for quantitative polymerase chain reaction (qPCR) analysis for the seven genes (Cyp2j5, Cyp2j6, Cyp2j8, Cyp2j9, Cyp2j11, Cyp2j12, and Cyp2j13) of the mouse Cyp2j subfamily. SYBR Green primer sets were developed and compared with commercially available TaqMan primer/probe assays for specificity toward mouse Cyp2j cDNA, and analysis of tissue distribution and regulation of Cyp2j genes. Each TaqMan primer/probe set and SYBR Green primer set were shown to be specific for their intended mouse Cyp2j cDNA. Tissue distribution of the mouse Cyp2j isoforms confirmed similar patterns of expression between the two qPCR methods. Cyp2j5 and Cyp2j13 were highly expressed in male kidneys, and Cyp2j11 was highly expressed in both male and female kidneys. Cyp2j6 was expressed in multiple tissues, with the highest expression in the small intestine and duodenum. Cyp2j8 was detected in various tissues, with highest expression found in the skin. Cyp2j9 was highly expressed in the brain, liver, and lung. Cyp2j12 was predominately expressed in the brain. We also determined the Cyp2j isoform expression in Cyp2j5 knockout mice to determine whether there was compensatory regulation of other Cyp2j isoforms, and we assessed Cyp2j isoform regulation during various inflammatory models, including influenza A, bacterial lipopolysaccharide, house dust mite allergen, and corn pollen. Both qPCR methods detected similar suppression of Cyp2j6 and Cyp2j9 during inflammation in the lung.


Assuntos
Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Animais , Sistema Enzimático do Citocromo P-450/biossíntese , Primers do DNA , DNA Complementar/biossíntese , DNA Complementar/genética , Feminino , Regulação Enzimológica da Expressão Gênica/genética , Hipersensibilidade/enzimologia , Hipersensibilidade/genética , Rim/enzimologia , Pulmão/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Infecções por Orthomyxoviridae/enzimologia , Pólen/imunologia , Reação em Cadeia da Polimerase , Distribuição Tecidual , Zea mays/imunologia
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