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1.
Biochem Biophys Res Commun ; 733: 150680, 2024 Nov 12.
Artículo en Inglés | MEDLINE | ID: mdl-39278094

RESUMEN

SCOPE: Triglyceride-based lipid emulsions are critical for total parenteral nutrition (TPN), but their long-term use has adverse effects, such as severe liver dysfunction necessitating improved formulations. This study compares the uptake mechanism and intracellular fate of novel glycerol-stabilized nano-sized lipid emulsions with conventional emulsions in CD4+ T cells, focusing on their impact on cellular metabolism. METHODS AND RESULTS: Nanoemulsions were formulated with increased glycerol content. Uptake of emulsions in primary human CD4+ T cells was investigated using different endocytic blockers, then quantified by flow cytometry, and visualized by confocal microscopy. To investigate emulsion intracellular fate, fatty acids in membrane phospholipids were quantified by GC-MS/MS and cellular metabolism was assessed by Seahorse technology. Results show T cells internalize both conventional and nano-sized emulsions using macropinocytosis. Fatty acids from emulsions are stored as neutral lipids in intracellular vesicles and are incorporated into phospholipids of cellular membranes. However, only nanoemulsions additionally use clathrin-mediated endocytosis and deliver fatty acids to mitochondria for increased ß-oxidation. CONCLUSIONS: Size of lipid emulsion droplets significantly influences their uptake and subsequent metabolism in CD4+ T cells. Our results highlight the potential for improved nutrient utilization with nanoemulsions in TPN formulations possibly leading to less adverse effects.


Asunto(s)
Linfocitos T CD4-Positivos , Emulsiones , Gotas Lipídicas , Humanos , Linfocitos T CD4-Positivos/metabolismo , Gotas Lipídicas/metabolismo , Tamaño de la Partícula , Endocitosis , Células Cultivadas , Ácidos Grasos/metabolismo , Triglicéridos/metabolismo
2.
Allergy ; 75(11): 2829-2845, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-32496587

RESUMEN

BACKGROUND: Morbidity and mortality from COVID-19 caused by novel coronavirus SARS-CoV-2 is accelerating worldwide, and novel clinical presentations of COVID-19 are often reported. The range of human cells and tissues targeted by SARS-CoV-2, its potential receptors and associated regulating factors are still largely unknown. The aim of our study was to analyze the expression of known and potential SARS-CoV-2 receptors and related molecules in the extensive collection of primary human cells and tissues from healthy subjects of different age and from patients with risk factors and known comorbidities of COVID-19. METHODS: We performed RNA sequencing and explored available RNA-Seq databases to study gene expression and co-expression of ACE2, CD147 (BSG), and CD26 (DPP4) and their direct and indirect molecular partners in primary human bronchial epithelial cells, bronchial and skin biopsies, bronchoalveolar lavage fluid, whole blood, peripheral blood mononuclear cells (PBMCs), monocytes, neutrophils, DCs, NK cells, ILC1, ILC2, ILC3, CD4+ and CD8+ T cells, B cells, and plasmablasts. We analyzed the material from healthy children and adults, and from adults in relation to their disease or COVID-19 risk factor status. RESULTS: ACE2 and TMPRSS2 were coexpressed at the epithelial sites of the lung and skin, whereas CD147 (BSG), cyclophilins (PPIA andPPIB), CD26 (DPP4), and related molecules were expressed in both epithelium and in immune cells. We also observed a distinct age-related expression profile of these genes in the PBMCs and T cells from healthy children and adults. Asthma, COPD, hypertension, smoking, obesity, and male gender status generally led to the higher expression of ACE2- and CD147-related genes in the bronchial biopsy, BAL, or blood. Additionally, CD147-related genes correlated positively with age and BMI. Interestingly, we also observed higher expression of CD147-related genes in the lesional skin of patients with atopic dermatitis. CONCLUSIONS: Our data suggest different receptor repertoire potentially involved in the SARS-CoV-2 infection at the epithelial barriers and in the immune cells. Altered expression of these receptors related to age, gender, obesity and smoking, as well as with the disease status, might contribute to COVID-19 morbidity and severity patterns.


Asunto(s)
Enzima Convertidora de Angiotensina 2/inmunología , Basigina/inmunología , COVID-19/epidemiología , Enfermedad Crónica/epidemiología , Dipeptidil Peptidasa 4/inmunología , SARS-CoV-2/inmunología , Adolescente , Adulto , Factores de Edad , Anciano , Enzima Convertidora de Angiotensina 2/genética , Asma/epidemiología , Asma/genética , Asma/inmunología , Basigina/genética , COVID-19/genética , COVID-19/inmunología , Niño , Preescolar , Comorbilidad , Dipeptidil Peptidasa 4/genética , Femenino , Expresión Génica/genética , Humanos , Hipertensión/epidemiología , Hipertensión/genética , Hipertensión/inmunología , Inmunidad Innata/inmunología , Lactante , Masculino , Persona de Mediana Edad , Obesidad/epidemiología , Obesidad/genética , Obesidad/inmunología , Enfermedad Pulmonar Obstructiva Crónica/epidemiología , Enfermedad Pulmonar Obstructiva Crónica/genética , Enfermedad Pulmonar Obstructiva Crónica/inmunología , Factores de Riesgo , SARS-CoV-2/genética , Adulto Joven
3.
Ann Allergy Asthma Immunol ; 125(5): 517-527, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-32454094

RESUMEN

OBJECTIVE: Our understanding of the origin of allergic diseases has increased in recent years, highlighting the importance of microbial dysbiosis and epithelial barrier dysfunction in affected tissues. Exploring the microbial-epithelial-immune crosstalk underlying the mechanisms of allergic diseases will allow the development of novel prevention and treatment strategies for allergic diseases. DATA SOURCES: This review summarizes the recent advances in microbial, epithelial, and immune interactions in atopic dermatitis, allergic rhinitis, chronic rhinosinusitis, and asthma. STUDY SELECTIONS: We performed a literature search, identifying relevant recent primary articles and review articles. RESULTS: Dynamic crosstalk between the environmental factors and microbial, epithelial, and immune cells in the development of atopic dermatitis, allergic rhinitis, chronic rhinosinusitis, and asthma underlies the pathogenesis of these diseases. There is substantial evidence in the literature suggesting that environmental factors directly affect barrier function of the epithelium. In addition, T-helper 2 (TH2) cells, type 2 innate lymphoid cells, and their cytokine interleukin 13 (IL-13) damage skin and lung barriers. The effects of environmental factors may at least in part be mediated by epigenetic mechanisms. Histone deacetylase activation by type 2 immune response has a major effect on leaky barriers and blocking of histone deacetylase activity corrects the defective barrier in human air-liquid interface cultures and mouse models of allergic asthma with rhinitis. We also present and discuss a novel device to detect and monitor skin barrier dysfunction, which provides an opportunity to rapidly and robustly assess disease severity. CONCLUSION: A complex interplay between environmental factors, epithelium, and the immune system is involved in the development of systemic allergic diseases.


Asunto(s)
Citocinas/inmunología , Epitelio/inmunología , Hipersensibilidad/inmunología , Hipersensibilidad/microbiología , Sistema Inmunológico/microbiología , Animales , Asma/inmunología , Asma/microbiología , Dermatitis Atópica/inmunología , Dermatitis Atópica/microbiología , Modelos Animales de Enfermedad , Epitelio/microbiología , Humanos , Inmunidad Innata , Linfocitos/inmunología , Ratones , Rinitis Alérgica/inmunología , Rinitis Alérgica/microbiología
4.
J Allergy Clin Immunol ; 144(5): 1242-1253.e7, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31082457

RESUMEN

BACKGROUND: A defective epithelial barrier is found in patients with allergic rhinitis (AR) and asthma; however, the underlying mechanisms remain poorly understood. Histone deacetylase (HDAC) activity has been identified as a crucial driver of allergic inflammation and tight junction dysfunction. OBJECTIVE: We investigated whether HDAC activity has been altered in patients with AR and in a mouse model of house dust mite (HDM)-induced allergic asthma and whether it contributed to epithelial barrier dysfunction. METHODS: Primary nasal epithelial cells of control subjects and patients with AR were cultured at the air-liquid interface to study transepithelial electrical resistance and paracellular flux of fluorescein isothiocyanate-dextran (4 kDa) together with mRNA expression and immunofluorescence staining of tight junctions. Air-liquid interface cultures were stimulated with different concentrations of JNJ-26481585, a broad-spectrum HDAC inhibitor. In vivo the effect of JNJ-26481585 on mucosal permeability and tight junction function was evaluated in a mouse model of HDM-induced allergic airway inflammation. RESULTS: General HDAC activity was greater in nasal epithelial cells of patients with AR and correlated inversely with epithelial integrity. Treatment of nasal epithelial cells with JNJ-26481585 restored epithelial integrity by promoting tight junction expression and protein reorganization. HDM-sensitized mice were treated with JNJ-26481585 to demonstrate the in vivo role of HDACs. Treated mice did not have allergic airway inflammation and had no bronchial hyperreactivity. Moreover, JNJ-26481585 treatment restored nasal mucosal function by promoting tight junction expression. CONCLUSION: Our findings identify increased HDAC activity as a potential tissue-injury mechanism responsible for dysregulated epithelial cell repair, leading to defective epithelial barriers in AR. Blocking HDAC activity is a promising novel target for therapeutic intervention in patients with airway diseases.


Asunto(s)
Asma/metabolismo , Histona Desacetilasas/metabolismo , Mucosa Nasal/metabolismo , Rinitis Alérgica/metabolismo , Uniones Estrechas/metabolismo , Animales , Antígenos Dermatofagoides/inmunología , Células Cultivadas , Modelos Animales de Enfermedad , Inhibidores de Histona Desacetilasas/farmacología , Humanos , Ácidos Hidroxámicos/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Mucosa Nasal/patología , Uniones Estrechas/patología
5.
J Allergy Clin Immunol ; 143(5): 1892-1903, 2019 05.
Artículo en Inglés | MEDLINE | ID: mdl-30500342

RESUMEN

BACKGROUND: Defects in the epithelial barrier have recently been associated with asthma and other allergies. The influence of laundry detergents on human bronchial epithelial cells (HBECs) and their barrier function remain unknown. OBJECTIVE: We investigated the effects of laundry detergents on cytotoxicity, barrier function, the transcriptome, and the epigenome in HBECs. METHODS: Air-liquid interface cultures of primary HBECs from healthy control subjects, patients with asthma, and patients with chronic obstructive pulmonary disease were exposed to laundry detergents and detergent residue after rinsing. Cytotoxicity and epithelial barrier function were evaluated. RNA sequencing, Assay for Transposase Accessible Chromatin with high-throughput sequencing, and DNA methylation arrays were used for checking the transcriptome and epigenome. RESULTS: Laundry detergents and rinse residue showed dose-dependent toxic effects on HBECs, with irregular cell shape and leakage of lactate dehydrogenase after 24 hours of exposure. A disrupted epithelial barrier function was found with decreased transepithelial electrical resistance, increased paracellular flux, and stratified tight junction (TJ) immunostaining in HBECs exposed to laundry detergent at 1:25,000 dilutions or rinse residue at further 1:10 dilutions. RNA sequencing analysis showed that lipid metabolism, apoptosis progress, and epithelially derived alarmin-related gene expression were upregulated, whereas cell adhesion-related gene expression was downregulated by laundry detergent at 1:50,000 dilutions after 24 hours of exposure without substantially affecting chromatin accessibility and DNA methylation. CONCLUSION: Our data demonstrate that laundry detergents, even at a very high dilution, and rinse residue show significant cell-toxic and directly disruptive effects on the TJ barrier integrity of HBECs without affecting the epigenome and TJ gene expression.


Asunto(s)
Células Epiteliales Alveolares/metabolismo , Asma/metabolismo , Bronquios/patología , Detergentes/metabolismo , Uniones Estrechas/metabolismo , Alarminas/genética , Alarminas/metabolismo , Células Epiteliales Alveolares/patología , Apoptosis , Células Cultivadas , Impedancia Eléctrica , Regulación de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , L-Lactato Deshidrogenasa/metabolismo , Metabolismo de los Lípidos , Uniones Estrechas/patología
6.
J Allergy Clin Immunol ; 143(6): 2190-2201.e9, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-30682454

RESUMEN

BACKGROUND: Group 2 innate lymphoid cells (ILC2s) play critical roles in induction and exacerbation of allergic airway inflammation. Thus clarification of the mechanisms that underlie regulation of ILC2 activation has received significant attention. Although innate lymphoid cells are divided into 3 major subsets that mirror helper effector T-cell subsets, counterpart subsets of regulatory T cells have not been well characterized. OBJECTIVE: We sought to determine the factors that induce regulatory innate lymphoid cells (ILCregs). METHODS: IL-10+ ILCregs induced from ILC2s by using retinoic acid (RA) were analyzed with RNA-sequencing and flow cytometry. ILCregs were evaluated in human nasal tissue from healthy subjects and patients with chronic rhinosinusitis with nasal polyps and lung tissue from house dust mite- or saline-treated mice. RESULTS: RA induced IL-10 secretion by human ILC2s but not type 2 cytokines. IL-10+ ILCregs, which were converted from ILC2s by means of RA stimulation, expressed a regulatory T cell-like signature with expression of IL-10, cytotoxic T lymphocyte-associated protein 4, and CD25, with downregulated effector type 2-related markers, such as chemoattractant receptor-homologous molecule on TH2 cells and ST2, and suppressed activation of CD4+ T cells and ILC2s. ILCregs were rarely detected in human nasal tissue from healthy subjects or lung tissue from saline-treated mice, but numbers were increased in nasal tissue from patients with chronic rhinosinusitis with nasal polyps and in lung tissue from house dust mite-treated mice. Enzymes for RA synthesis were upregulated in airway epithelial cells during type 2 inflammation in vivo and by IL-13 in vitro. CONCLUSION: We have identified a unique immune regulatory and anti-inflammatory pathway by which RA converts ILC2s to ILCregs. Interactions between airway epithelial cells and ILC2s play an important roles in the generation of ILCregs.


Asunto(s)
Antiinflamatorios/farmacología , Linfocitos/efectos de los fármacos , Tretinoina/farmacología , Animales , Línea Celular , Citocinas/inmunología , Células Epiteliales/inmunología , Humanos , Inmunidad Innata , Pulmón/inmunología , Linfocitos/inmunología , Ratones Endogámicos C57BL , Senos Paranasales/inmunología
7.
Allergy ; 74(10): 1934-1944, 2019 10.
Artículo en Inglés | MEDLINE | ID: mdl-30989659

RESUMEN

BACKGROUND: Many skin and mucosal inflammatory disorders, such as atopic dermatitis, have been associated with an impaired epithelial barrier function, which allows allergens, pollutants, or microbes to enter the tissue and activate the immune response. The aim of this study was to establish a method to directly assess in vivo the epidermal barrier function by electrical impedance (EI) spectroscopy. METHODS: Mice epidermal barrier was damaged by epicutaneous application of proteases and cholera toxin and by tape stripping. EI and transepidermal water loss (TEWL) were measured before and after the application. Additionally, histological analysis, immunofluorescence staining, and RT-PCR were performed on skin biopsies to evaluate the epithelial barrier. RESULTS: A few hours after papain application, a dose-dependent reduction of EI was detected, reflecting the decreased barrier function. At the same time, an increase of TEWL was observed, with a significant negative correlation with EI, demonstrating that EI changes were directly linked to barrier defects. Twenty-four and 48 hours after the treatment, EI starts to increase to background levels, indicating tissue healing and restoration of skin barrier. Barrier disruption was confirmed by histological analysis showing an impaired stratum corneum and higher cellular infiltration after papain application. In addition, immunofluorescence staining and RT-PCR showed downregulation of molecules involved in the barrier function, such as filaggrin, occludin, and claudin-1, and mRNA levels of filaggrin, loricrin, and involucrin. Comparable results were observed after tape stripping and cholera toxin treatment. CONCLUSION: Electrical impedance spectroscopy is a rapid and reliable diagnostic tool to detect skin barrier defects.


Asunto(s)
Espectroscopía Dieléctrica , Epidermis/fisiología , Fenómenos Fisiológicos de la Piel , Animales , Biopsia , Modelos Animales de Enfermedad , Proteínas Filagrina , Humanos , Ratones , Pérdida Insensible de Agua
8.
J Allergy Clin Immunol ; 141(3): 951-963.e8, 2018 03.
Artículo en Inglés | MEDLINE | ID: mdl-29074456

RESUMEN

BACKGROUND: Allergic rhinitis (AR) is characterized by mucosal inflammation, driven by activated immune cells. Mast cells and TH2 cells might decrease epithelial barrier integrity in AR, maintaining a leaky epithelial barrier. OBJECTIVE: We sought to investigate the role of histamine and TH2 cells in driving epithelial barrier dysfunction in AR. METHODS: Air-liquid interface cultures of primary nasal epithelial cells were used to measure transepithelial electrical resistance, paracellular flux of fluorescein isothiocyanate-dextran 4 kDa, and mRNA expression of tight junctions. Nasal secretions were collected from healthy control subjects, AR patients, and idiopathic rhinitis patients and were tested in vitro. In addition, the effect of activated TH1 and TH2 cells, mast cells, and neurons was tested in vitro. The effect of IL-4, IL-13, IFN-γ, and TNF-α on mucosal permeability was tested in vivo. RESULTS: Histamine as well as nasal secretions of AR but not idiopathic rhinitis patients rapidly decreased epithelial barrier integrity in vitro. Pretreatment with histamine receptor-1 antagonist, azelastine prevented the early effect of nasal secretions of AR patients on epithelial integrity. Supernatant of activated TH1 and TH2 cells impaired epithelial integrity, while treatment with anti-TNF-α or anti-IL-4Rα monoclonal antibodies restored the TH1- and TH2-induced epithelial barrier dysfunction, respectively. IL-4, IFN-γ, and TNF-α enhanced mucosal permeability in mice. Antagonizing IL-4 prevented mucosal barrier disruption and tight junction downregulation in a mouse model of house dust mite allergic airway inflammation. CONCLUSIONS: Our data indicate a key role for allergic inflammatory mediators in modulating nasal epithelial barrier integrity in the pathophysiology in AR.


Asunto(s)
Citocinas/inmunología , Histamina/inmunología , Mucosa Nasal/inmunología , Rinitis Alérgica/inmunología , Células TH1/inmunología , Células Th2/inmunología , Animales , Línea Celular , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Mucosa Nasal/patología , Rinitis Alérgica/patología , Células TH1/patología , Células Th2/patología
9.
J Allergy Clin Immunol ; 141(1): 300-310.e11, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-28392332

RESUMEN

BACKGROUND: Bronchial epithelial barrier leakiness and type 2 innate lymphoid cells (ILC2s) have been separately linked to asthma pathogenesis; however, the influence of ILC2s on the bronchial epithelial barrier has not been investigated previously. OBJECTIVE: We investigated the role of ILC2s in the regulation of bronchial epithelial tight junctions (TJs) and barrier function both in bronchial epithelial cells of asthmatic patients and healthy subjects and general innate lymphoid cell- and ILC2-deficient mice. METHODS: Cocultures of human ILC2s and bronchial epithelial cells were used to determine transepithelial electrical resistance, paracellular flux, and TJ mRNA and protein expressions. The effect of ILC2s on TJs was examined by using a murine model of IL-33-induced airway inflammation in wild-type, recombination-activating gene 2 (Rag2)-/-, Rag2-/-Il2rg-/-, and Rorasg/sg mice undergoing bone marrow transplantation to analyze the in vivo relevance of barrier disruption by ILC2s. RESULTS: ILC2s significantly impaired the epithelial barrier, as demonstrated by reduced transepithelial electrical resistance and increased fluorescein isothiocyanate-dextran permeability in air-liquid interface cultures of human bronchial epithelial cells. This was in parallel to decreased mRNAs and disrupted protein expression of TJ proteins and was restored by neutralization of IL-13. Intranasal administration of recombinant IL-33 to wild-type and Rag2-/- mice lacking T and B cells triggered TJ disruption, whereas Rag2-/-Il2rg-/- and Rorasg/sg mice undergoing bone marrow transplantation that lack ILC2s did not show any barrier leakiness. Direct nasal administration of IL-13 was sufficient to induce deficiency in the TJ barrier in the bronchial epithelium of mice in vivo. CONCLUSION: These data highlight an essential mechanism in asthma pathogenesis by demonstrating that ILC2s are responsible for bronchial epithelial TJ barrier leakiness through IL-13.


Asunto(s)
Asma/inmunología , Asma/metabolismo , Inmunidad Innata , Interleucina-13/metabolismo , Subgrupos Linfocitarios/inmunología , Subgrupos Linfocitarios/metabolismo , Mucosa Respiratoria/inmunología , Mucosa Respiratoria/metabolismo , Uniones Estrechas/metabolismo , Animales , Modelos Animales de Enfermedad , Células Epiteliales/metabolismo , Humanos , Interleucina-13/antagonistas & inhibidores , Ratones , Ratones Transgénicos , Moco/metabolismo , Mucosa Respiratoria/patología
10.
J Allergy Clin Immunol ; 139(1): 93-103, 2017 01.
Artículo en Inglés | MEDLINE | ID: mdl-27312821

RESUMEN

BACKGROUND: Tight junctions (TJs) form a barrier on the apical side of neighboring epithelial cells in the bronchial mucosa. Changes in their integrity might play a role in asthma pathogenesis by enabling the paracellular influx of allergens, toxins, and microbes to the submucosal tissue. OBJECTIVE: The regulation of bronchial epithelial TJs by TH2 cells and their cytokines and their involvement in epigenetic regulation of barrier function were investigated. METHODS: The expression, regulation, and function of TJs were determined in air-liquid interface (ALI) cultures of control and asthmatic primary human bronchial epithelial cells (HBECs) by means of analysis of transepithelial electrical resistance, paracellular flux, mRNA expression, Western blotting, and immunofluorescence staining. RESULTS: HBECs from asthmatic patients showed a significantly low TJ integrity in ALI cultures compared with HBECs from healthy subjects. TH2 cell numbers and levels of their cytokines, IL-4 and IL-13, decreased barrier integrity in ALI cultures of HBECs from control subjects but not in HBECs from asthmatic patients. They induced a physical separation of the TJs of adjacent cells in immunofluorescence staining of the TJ molecules occludin and zonula occludens-1. We observed that expression of histone deacetylases (HDACs) 1 and 9, and Silent information regulator genes (sirtuins [SIRTs]) 6 and 7 were significantly high in HBECs from asthmatic patients. IL-4 and IL-13 significantly increased the expression of HDACs and SIRTs. The role of HDAC activation on epithelial barrier leakiness was confirmed by HDAC inhibition, which improved barrier integrity through increased synthesis of TJ molecules in epithelium from asthmatic patients to the level seen in HBECs from control subjects. CONCLUSION: Our data demonstrate that barrier leakiness in asthmatic patients is induced by TH2 cells, IL-4, and IL-13 and HDAC activity. The inhibition of endogenous HDAC activity reconstitutes defective barrier by increasing TJ expression.


Asunto(s)
Asma/metabolismo , Citocinas/metabolismo , Células Epiteliales/metabolismo , Uniones Estrechas/metabolismo , Adulto , Bronquios/citología , Femenino , Histona Desacetilasas/genética , Histona Desacetilasas/metabolismo , Humanos , Masculino , Persona de Mediana Edad , ARN Mensajero/metabolismo , Células Th2/metabolismo
11.
J Allergy Clin Immunol ; 137(6): 1681-1696, 2016 06.
Artículo en Inglés | MEDLINE | ID: mdl-27090934

RESUMEN

This review highlights some of the advances in mechanisms of allergic disease, particularly anaphylaxis, including food allergy, drug hypersensitivity, atopic dermatitis (AD), allergic conjunctivitis, and airway diseases. During the last year, a mechanistic advance in food allergy was achieved by focusing on mechanisms of allergen sensitization. Novel biomarkers and treatment for mastocytosis were presented in several studies. Novel therapeutic approaches in the treatment of atopic dermatitis and psoriasis showed that promising supplementation of the infant's diet in the first year of life with immunoactive prebiotics might have a preventive role against early development of AD and that therapeutic approaches to treat AD in children might be best directed to the correction of a TH2/TH1 imbalance. Several studies were published emphasizing the role of the epithelial barrier in patients with allergic diseases. An impaired skin barrier as a cause for sensitization to food allergens in children and its relationship to filaggrin mutations has been an important development. Numerous studies presented new approaches for improvement of epithelial barrier function and novel biologicals used in the treatment of inflammatory skin and eosinophilic diseases. In addition, novel transcription factors and signaling molecules that can develop as new possible therapeutic targets have been reported.


Asunto(s)
Hipersensibilidad/etiología , Hipersensibilidad/metabolismo , Alérgenos/administración & dosificación , Alérgenos/inmunología , Anafilaxia/diagnóstico , Anafilaxia/epidemiología , Anafilaxia/etiología , Anafilaxia/metabolismo , Animales , Conjuntivitis Alérgica/etiología , Conjuntivitis Alérgica/metabolismo , Dermatitis Atópica/etiología , Dermatitis Atópica/metabolismo , Desensibilización Inmunológica , Proteínas Filagrina , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/epidemiología , Hipersensibilidad a los Alimentos/etiología , Hipersensibilidad a los Alimentos/metabolismo , Humanos , Hipersensibilidad/diagnóstico , Hipersensibilidad/epidemiología , Tolerancia Inmunológica , Huésped Inmunocomprometido/genética , Huésped Inmunocomprometido/inmunología , Mastocitosis/etiología , Mastocitosis/metabolismo
12.
J Allergy Clin Immunol ; 137(4): 1043-1053.e5, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26846377

RESUMEN

BACKGROUND: Tight junction (TJ) defects have recently been associated with asthma and chronic rhinosinusitis. The expression, function, and regulation of nasal epithelial TJs remain unknown in patients with allergic rhinitis (AR). OBJECTIVE: We investigated the expression, function, and regulation of TJs in the nasal epithelium of patients with house dust mite (HDM)-induced AR and in an HDM-induced murine model of allergic airway disease. METHODS: Air-liquid interface cultures of primary nasal epithelial cells of control subjects and patients with HDM-induced AR were used for measuring transepithelial resistance and passage to fluorescein isothiocyanate-dextran 4 kDa (FD4). Ex vivo transtissue resistance and FD4 permeability of nasal mucosal explants were measured. TJ expression was evaluated by using real-time quantitative PCR and immunofluorescence. In addition, the effects of IL-4, IFN-γ, and fluticasone propionate (FP) on nasal epithelial cells were investigated in vitro. An HDM murine model was used to study the effects of allergic inflammation and FP treatment on transmucosal passage of FD4 in vivo. RESULTS: A decreased resistance in vitro and ex vivo was found in patients with HDM-induced AR, with increased FD4 permeability and reduced occludin and zonula occludens-1 expression. AR symptoms correlated inversely with resistance in patients with HDM-induced AR. In vitro IL-4 decreased transepithelial resistance and increased FD4 permeability, whereas IFN-γ had no effect. FP prevented IL-4-induced barrier dysfunction in vitro. In an HDM murine model FP prevented the allergen-induced increased mucosal permeability. CONCLUSION: We found impaired nasal epithelial barrier function in patients with HDM-induced AR, with lower occludin and zonula occludens-1 expression. IL-4 disrupted epithelial integrity in vitro, and FP restored barrier function. Better understanding of nasal barrier regulation might lead to a better understanding and treatment of AR.


Asunto(s)
Mucosa Nasal/metabolismo , Ocludina/metabolismo , Pyroglyphidae/inmunología , Rinitis Alérgica Perenne/metabolismo , Uniones Estrechas/metabolismo , Proteína de la Zonula Occludens-1/metabolismo , Adulto , Animales , Antiinflamatorios/uso terapéutico , Biomarcadores/metabolismo , Estudios de Casos y Controles , Dextranos/metabolismo , Femenino , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Fluticasona/uso terapéutico , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Mucosa Nasal/inmunología , Permeabilidad , Reacción en Cadena en Tiempo Real de la Polimerasa , Rinitis Alérgica Perenne/tratamiento farmacológico , Rinitis Alérgica Perenne/inmunología
13.
J Allergy Clin Immunol ; 138(4): 984-1010, 2016 10.
Artículo en Inglés | MEDLINE | ID: mdl-27577879

RESUMEN

There have been extensive developments on cellular and molecular mechanisms of immune regulation in allergy, asthma, autoimmune diseases, tumor development, organ transplantation, and chronic infections during the last few years. Better understanding the functions, reciprocal regulation, and counterbalance of subsets of immune and inflammatory cells that interact through interleukins, interferons, TNF-α, and TGF-ß offer opportunities for immune interventions and novel treatment modalities in the era of development of biological immune response modifiers particularly targeting these molecules or their receptors. More than 60 cytokines have been designated as interleukins since the initial discoveries of monocyte and lymphocyte interleukins (called IL-1 and IL-2, respectively). Studies of transgenic or gene-deficient mice with altered expression of these cytokines or their receptors and analyses of mutations and polymorphisms in human genes that encode these products have provided essential information about their functions. Here we review recent developments on IL-1 to IL-38, TNF-α, TGF-ß, and interferons. We highlight recent advances during the last few years in this area and extensively discuss their cellular sources, targets, receptors, signaling pathways, and roles in immune regulation in patients with allergy and asthma and other inflammatory diseases.


Asunto(s)
Enfermedades del Sistema Inmune , Interferones/fisiología , Interleucinas/fisiología , Linfocitos T Reguladores/inmunología , Factor de Crecimiento Transformador beta/fisiología , Factor de Necrosis Tumoral alfa/fisiología , Animales , Humanos
16.
J Allergy Clin Immunol ; 130(5): 1087-1096.e10, 2012 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-22840853

RESUMEN

BACKGROUND: Chronic rhinosinusitis (CRS) is a common disease with still unclear pathophysiologic mechanisms. Epithelial tight junctions (TJs) have been shown to be involved in different chronic disorders, including bronchial asthma, inflammatory bowel diseases, and skin disorders. The regulation of epithelial barrier function and TJ expression has not been extensively studied in patients with CRS and in the paranasal sinus epithelium thus far. OBJECTIVE: We sought to elucidate the TJ expression pattern in the epithelium of the sinonasal mucosa and its regulation in patients with CRS. METHODS: Trans-tissue resistance was measured in biopsy specimens from healthy control subjects and patients with CRS with and without nasal polyps. TJ protein expression was determined by using immunofluorescence, Western blotting, and real-time PCR. Primary epithelial cell cultures from patients with CRS and control subjects were used in air-liquid interface (ALI) cultures for the measurement of transepithelial resistance (TER) and TJ expression. The effect of IFN-γ, IL-4, and IL-17 on ALI cultures was assessed. RESULTS: A decreased trans-tissue resistance was found in biopsy specimens from patients with CRS with nasal polyps along with an irregular, patchy, and decreased expression of the TJ molecules occludin and zonula occludens 1. TER was reduced in ALI cultures from patients with CRS with nasal polyps. The cytokines IFN-γ and IL-4 decreased TER, whereas IL-17 did not have any influence on epithelial integrity. CONCLUSION: A defective epithelial barrier was found in patients with CRS with nasal polyps along with a decreased expression of TJ proteins. The disruption of epithelial integrity by IFN-γ and IL-4 in vitro indicates a possible role for these proinflammatory cytokines in the pathogenesis of patients with CRS.


Asunto(s)
Mucosa Nasal/inmunología , Pólipos Nasales/inmunología , Rinitis/inmunología , Sinusitis/inmunología , Uniones Estrechas/inmunología , Biopsia , Células Cultivadas , Enfermedad Crónica , Claudina-4/genética , Claudina-4/metabolismo , Regulación de la Expresión Génica , Humanos , Interferón gamma/inmunología , Interleucina-17/inmunología , Interleucina-4/inmunología , Pólipos Nasales/complicaciones , Pólipos Nasales/patología , Ocludina/genética , Ocludina/metabolismo , Cultivo Primario de Células , Rinitis/complicaciones , Rinitis/patología , Sinusitis/complicaciones , Sinusitis/patología , Uniones Estrechas/patología , Proteína de la Zonula Occludens-1/genética , Proteína de la Zonula Occludens-1/metabolismo
17.
J Allergy Clin Immunol ; 129(4): 964-73.e7, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22336080

RESUMEN

BACKGROUND: IL-32 is a proinflammatory cytokine involved in various chronic inflammatory diseases. Chronic airway inflammation in asthmatic patients results in structural airway changes, including angiogenesis. Vascular endothelial growth factor (VEGF) is a key inducer of angiogenesis in the airways of asthmatic patients. OBJECTIVE: The aim of the study was to investigate the expression and function of IL-32 in patients with angiogenesis and asthma. METHODS: The expression and regulation of IL-32 in normal human bronchial epithelial (NHBE) cells was analyzed by using RT-PCR, ELISA, Western blotting, immunofluorescent staining, and flow cytometry. After knockdown of IL-32 in NHBE cells by small interfering RNA (siRNA) transfections, VEGF secretion was quantified by means of ELISA. New blood vessel formation was determined with human umbilical vein endothelial cells by culturing with supernatants from IL-32 siRNA-transfected NHBE cells. IL-32 was determined in serum and induced sputum samples of asthmatic patients and healthy control subjects by means of ELISA. RESULTS: IL-32 is expressed in NHBE cells on stimulation with IFN-γ, TNF-α, T(H)1 cells, and rhinovirus. Inhibition of IL-32 expression resulted in significantly increased secretion of the proangiogenic factors VEGF and platelet-derived growth factor by NHBE cells. Human umbilical vein endothelial cells cultured in supernatants from IL-32 siRNA-transfected NHBE cells showed enhanced in vitro angiogenesis. IL-32 is detectable in induced sputum from asthmatic patients. IL-32 serum levels were significantly higher in asthmatic patients compared with those seen in healthy control subjects and correlated with response to asthma treatment. CONCLUSION: IL-32 is induced by IFN-γ, TNF-α, T(H)1 cells, and rhinovirus in bronchial epithelial cells. It inhibits angiogenesis, and its serum levels are associated with a good treatment response in asthmatic patients.


Asunto(s)
Asma/metabolismo , Bronquios/irrigación sanguínea , Interleucinas/metabolismo , Neovascularización Patológica/metabolismo , Adolescente , Adulto , Anciano , Antiasmáticos/uso terapéutico , Asma/tratamiento farmacológico , Asma/genética , Línea Celular , Células Cultivadas , Células Epiteliales/metabolismo , Femenino , Silenciador del Gen , Humanos , Interferón gamma/sangre , Interleucinas/genética , Masculino , Persona de Mediana Edad , Neovascularización Patológica/genética , Infecciones por Picornaviridae/inmunología , Infecciones por Picornaviridae/metabolismo , Factor de Crecimiento Derivado de Plaquetas/metabolismo , ARN Mensajero/metabolismo , Mucosa Respiratoria/metabolismo , Receptores Toll-Like/metabolismo , Factor de Necrosis Tumoral alfa/sangre , Regulación hacia Arriba , Factor A de Crecimiento Endotelial Vascular/metabolismo , Adulto Joven
18.
Mucosal Immunol ; 16(1): 5-16, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-36642382

RESUMEN

SARS-CoV-2 enters human cells through its main receptor, angiotensin-converting enzyme 2 (ACE2), which constitutes a limiting factor of infection. Recent findings demonstrating novel ACE2 isoforms implicate that this receptor is regulated in a more complex way than previously anticipated. However, it remains unknown how various inflammatory conditions influence the abundance of these ACE2 variants. Hence, we studied expression of ACE2 messenger RNA (mRNA) and protein isoforms, together with its glycosylation and spatial localization in primary human airway epithelium upon allergic inflammation and viral infection. We found that interleukin-13, the main type 2 cytokine, decreased expression of long ACE2 mRNA and reduced glycosylation of full-length ACE2 protein via alteration of N-linked glycosylation process, limiting its availability on the apical side of ciliated cells. House dust mite allergen did not affect the expression of ACE2. Rhinovirus infection increased short ACE2 mRNA, but it did not influence its protein expression. In addition, by screening other SARS-CoV-2 related host molecules, we found that interleukin-13 and rhinovirus significantly regulated mRNA, but not protein of transmembrane serine protease 2 and neuropilin 1. Regulation of ACE2 and other host proteins was comparable in healthy and asthmatic epithelium, underlining the lack of intrinsic differences but dependence on the inflammatory milieu in the airways.


Asunto(s)
Enzima Convertidora de Angiotensina 2 , COVID-19 , Humanos , SARS-CoV-2/metabolismo , Interleucina-13 , Peptidil-Dipeptidasa A/genética , Inflamación , Epitelio/metabolismo , ARN Mensajero/metabolismo , Isoformas de Proteínas
19.
Clin Nutr ; 42(12): 2422-2433, 2023 12.
Artículo en Inglés | MEDLINE | ID: mdl-37871483

RESUMEN

BACKGROUND AND AIMS: Parenteral nutrition (PN) rich in n-6 and n-3 long-chain fatty acids is used in clinical practice for nourishing patients who are unable to receive adequate nutrition through their digestive systems. In this study, we compare the effect on inflammation of the commonly used lipid emulsions Omegaven (n-3-rich) and Intralipid (n-6-rich) in human peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs were treated with different doses of n-3-rich Omegaven and n-6-rich Intralipid and the immune cells were characterized by flow cytometry. RESULTS: We show that incubation of PBMCs with n-3-rich Omegaven leads to an increase in expression of CD1d and CD86 in CD14+monocytes. At the same time, an increased number of NKT cells expressing cytotoxic T cell antigen 4 is observed, suggesting immunological synapse formation. Both CD14+monocytes and NKT cells showed an increase in IL-10 production and a reduction in the pro-inflammatory cytokines IFN-γ, TNF-α, and IL-4, which led to an increase in the number of FOXP3+T regulatory cells. In addition, we show that n-3-rich Omegaven reduces the expression of TNFα, IFNγ and IL-4 in CD4+T and CD8+T cells independent of the presented interaction between CD14+monocytes and NKT cells. The described mechanism of n-3 rich lipid emulsions was confirmed in PBMCs from patients with inflammatory bowel disease but not in colorectal cancer patients which seem to lack the interaction between CD14+monocytes and NKT cells. CONCLUSIONS: These results show a mechanism for the beneficial effect of the n-3-rich Omegaven in patients with inflammatory conditions but questions its use in patients with cancer. Hence, our results may assist in choosing the best lipid emulsion for patients who require PN.


Asunto(s)
Ácidos Grasos Omega-3 , Humanos , Ácidos Grasos Omega-3/farmacología , Emulsiones/farmacología , Interleucina-4 , Leucocitos Mononucleares/metabolismo , Nutrición Parenteral/métodos , Factor de Necrosis Tumoral alfa/metabolismo , Antiinflamatorios
20.
Nat Commun ; 14(1): 2329, 2023 04 22.
Artículo en Inglés | MEDLINE | ID: mdl-37087523

RESUMEN

Rhinoviruses and allergens, such as house dust mite are major agents responsible for asthma exacerbations. The influence of pre-existing airway inflammation on the infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is largely unknown. We analyse mechanisms of response to viral infection in experimental in vivo rhinovirus infection in healthy controls and patients with asthma, and in in vitro experiments with house dust mite, rhinovirus and SARS-CoV-2 in human primary airway epithelium. Here, we show that rhinovirus infection in patients with asthma leads to an excessive RIG-I inflammasome activation, which diminishes its accessibility for type I/III interferon responses, leading to their early functional impairment, delayed resolution, prolonged viral clearance and unresolved inflammation in vitro and in vivo. Pre-exposure to house dust mite augments this phenomenon by inflammasome priming and auxiliary inhibition of early type I/III interferon responses. Prior infection with rhinovirus followed by SARS-CoV-2 infection augments RIG-I inflammasome activation and epithelial inflammation. Timely inhibition of the epithelial RIG-I inflammasome may lead to more efficient viral clearance and lower the burden of rhinovirus and SARS-CoV-2 infections.


Asunto(s)
Factores de Restricción Antivirales , Asma , COVID-19 , Proteína 58 DEAD Box , Inflamasomas , Rhinovirus , Humanos , Factores de Restricción Antivirales/genética , Factores de Restricción Antivirales/metabolismo , Asma/genética , Asma/inmunología , COVID-19/genética , COVID-19/inmunología , Proteína 58 DEAD Box/metabolismo , Infecciones por Enterovirus/genética , Infecciones por Enterovirus/inmunología , Inflamasomas/genética , Inflamasomas/metabolismo , Inflamación , Interferón Tipo I , Infecciones por Picornaviridae/genética , Infecciones por Picornaviridae/inmunología , Rhinovirus/metabolismo , Rhinovirus/patogenicidad , SARS-CoV-2
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