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Nariz , Humanos , Mucosa Nasal/imunologia , Mucosa Nasal/citologia , Nariz/citologia , Nariz/imunologiaRESUMO
Nasal vaccination elicits a humoral immune response that provides protection from airborne pathogens1, yet the origins and specific immune niches of antigen-specific IgA-secreting cells in the upper airways are unclear2. Here we define nasal glandular acinar structures and the turbinates as immunological niches that recruit IgA-secreting plasma cells from the nasal-associated lymphoid tissues (NALTs)3. Using intact organ imaging, we demonstrate that nasal vaccination induces B cell expansion in the subepithelial dome of the NALT, followed by invasion into commensal-bacteria-driven chronic germinal centres in a T cell-dependent manner. Initiation of the germinal centre response in the NALT requires pre-expansion of antigen-specific T cells, which interact with cognate B cells in interfollicular regions. NALT ablation and blockade of PSGL-1, which mediates interactions with endothelial cell selectins, demonstrated that NALT-derived IgA-expressing B cells home to the turbinate region through the circulation, where they are positioned primarily around glandular acinar structures. CCL28 expression was increased in the turbinates in response to vaccination and promoted homing of IgA+ B cells to this site. Thus, in response to nasal vaccination, the glandular acini and turbinates provide immunological niches that host NALT-derived IgA-secreting cells. These cellular events could be manipulated in vaccine design or in the treatment of upper airway allergic responses.
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Imunoglobulina A , Tecido Linfoide , Mucosa Nasal , Plasmócitos , Linfócitos T , Conchas Nasais , Animais , Feminino , Masculino , Camundongos , Bactérias/imunologia , Movimento Celular , Quimiocinas CC/imunologia , Quimiocinas CC/metabolismo , Centro Germinativo/imunologia , Centro Germinativo/citologia , Imunoglobulina A/imunologia , Imunoglobulina A/metabolismo , Tecido Linfoide/imunologia , Tecido Linfoide/citologia , Camundongos Endogâmicos C57BL , Mucosa Nasal/citologia , Mucosa Nasal/imunologia , Plasmócitos/imunologia , Plasmócitos/citologia , Plasmócitos/metabolismo , Linfócitos T/imunologia , Linfócitos T/citologia , Linfócitos T/metabolismo , Conchas Nasais/citologia , Conchas Nasais/imunologia , Vacinação , Administração Intranasal , Vacinas/imunologia , SimbioseRESUMO
The function of a well-differentiated nasal epithelium is largely affected by airflow-induced wall shear stress, yet fewin vitromodels recapitulate this dynamic condition. Models which do expose cells to airflow exclusively initiate flow after the differentiation process has occurred.In vivo, basal cells are constantly replenishing the epithelium under airflow conditions, indicating that airflow may affect the development and function of the differentiated epithelium. To address this gap in the field, we developed a physiologically relevant microphysiological model of the human nasal epithelium and investigated the effects of exposing cells to airflow during epithelial maturation at the air-liquid interface. The nasal airway-on-chip platform was engineered to mimic bi-directional physiological airflow during normal breathing. Primary human nasal epithelial cells were seeded on chips and subjected to either: (1) no flow, (2) single flow (0.5 dyne cm-2flow on Day 21 of ALI only), or (3) pre-conditioning flow (0.05 dyne cm-2on Days 14-20 and 0.5 dyne cm-2flow on Day 21) treatments. Cells exposed to pre-conditioning showed decreased morphological changes and mucus secretions, as well as decreased inflammation, compared to unconditioned cells. Our results indicate that flow exposure only post-differentiation may impose acute stress on cells, while pre-conditioning may potentiate a properly functioning epitheliumin vitro.
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Células Epiteliais , Dispositivos Lab-On-A-Chip , Mucosa Nasal , Humanos , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Mucosa Nasal/citologia , Mucosa Nasal/metabolismo , Diferenciação Celular , Células Cultivadas , Ar , Modelos Biológicos , Engenharia TecidualRESUMO
Sepsis is characterized by an exacerbated inflammatory response, driven by the overproduction of cytokines, a phenomenon known as a cytokine storm. This condition is further compounded by the extensive infiltration of M1 macrophages and the pyroptosis of these cells, leading to immune paralysis. To counteract this, we sought to transition M1 macrophages into the M2 phenotype and safeguard them from pyroptosis. For this purpose, we employed ectodermal mesenchymal stem cells (EMSCs) sourced from the nasal mucosa to examine their impact on both macrophages and septic animal models. The co-culture protocol involving LPS-stimulated rat bone marrow macrophages and EMSCs was employed to examine the paracrine influence of EMSCs on macrophages. The intravenous administration of EMSCs was utilized to observe the enhancement in the survival rate of septic rat models and the protection of associated organs. The findings indicated that EMSCs facilitated M2 polarization of macrophages, which were stimulated by LPS, and significantly diminished levels of pro-inflammatory cytokines and NLRP3. Furthermore, EMSCs notably restored the mitochondrial membrane potential (MMP) of macrophages through paracrine action, eliminated excess reactive oxygen species (ROS), and inhibited macrophage pyroptosis. Additionally, the systemic integration of EMSCs substantially reduced injuries to multiple organs and preserved the fundamental functions of the heart, liver, and kidney in CLP rats, thereby extending their survival.
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Macrófagos , Células-Tronco Mesenquimais , Mucosa Nasal , Piroptose , Sepse , Animais , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Ratos , Mucosa Nasal/imunologia , Mucosa Nasal/citologia , Sepse/imunologia , Masculino , Ratos Sprague-Dawley , Transplante de Células-Tronco Mesenquimais/métodos , Lipopolissacarídeos , Citocinas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Modelos Animais de Doenças , Técnicas de Cocultura , Potencial da Membrana Mitocondrial , Células CultivadasRESUMO
Plants offer a cost-effective and scalable pharmaceutical platform devoid of host-derived contamination risks. However, their medical application is complicated by the potential for acute allergic reactions to external proteins. Developing plant-based protein therapeutics for localized diseases with non-invasive treatment modalities may capitalize on the benefits of plant proteins while avoiding their inherent risks. Dupilumab, which is effective against a variety of allergic and autoimmune diseases but has systemic responses and injection-related side effects, may be more beneficial if delivered locally using a small biological form. In this study, we engineered a single-chain variable fragment (scFv) of dupilumab, termed Dup-scFv produced by Nicotiana benthamiana, and evaluated its tissue permeability and anti-inflammatory efficacy in air-liquid interface cultured human nasal epithelial cells (HNECs). Despite showing 3.67- and 17-fold lower binding affinity for IL-4Ra in surface plasmon resonance assays and cell binding assays, respectively, Dup-scFv retained most of the affinity of dupilumab, which was originally high, with a dissociation constant (KD) of 4.76 pM. In HNECs cultured at the air-liquid interface, Dup-scFv administered on the air side inhibited the inflammatory marker CCL26 in hard-to-reach basal cells more effectively than dupilumab. In addition, Dup-scFv had an overall permeability of 0.8% across cell layers compared to undetectable levels of dupilumab. These findings suggest that plant-produced Dup-scFv can be delivered non-invasively to cultured HNESc to alleviate inflammatory signaling, providing a practical approach to utilize plant-based proteins for topical therapeutic applications.
Assuntos
Anticorpos Monoclonais Humanizados , Células Epiteliais , Nicotiana , Anticorpos de Cadeia Única , Humanos , Nicotiana/metabolismo , Anticorpos Monoclonais Humanizados/farmacologia , Células Epiteliais/metabolismo , Células Epiteliais/efeitos dos fármacos , Anticorpos de Cadeia Única/farmacologia , Anticorpos de Cadeia Única/genética , Quimiocinas CC/metabolismo , Subunidade alfa de Receptor de Interleucina-4/metabolismo , Células Cultivadas , Mucosa Nasal/metabolismo , Mucosa Nasal/citologia , Mucosa Nasal/imunologiaRESUMO
BACKGROUND: Chronic rhinosinusitis (CRS) is an inflammatory disease affecting the sinuses or nose. Persistent inflammatory responses can lead to tissue remodeling, which is a pathological characteristics of CRS. Activation of fibroblasts in the nasal mucosal stroma, differentiation and collagen deposition, and subepithelial fibrosis have been associated with CRS. OBJECTIVES: We aimed to assess the inhibitory effects of doxycycline and deoxycholic acid-polyethyleneimine conjugate (DA3-Doxy) on myofibroblast differentiation and extracellular matrix (ECM) production in nasal fibroblasts stimulated with TGF-ß1. METHODS: To enhance efficacy, we prepared DA3-Doxy using a conjugate of low-molecular-weight polyethyleneimine (PEI) (MW 1800) and deoxycholic acid (DA) and Doxy. The synthesis of the DA3-Doxy polymer was confirmed using nuclear magnetic resonance, and the critical micelle concentration required for cationic micelle formation through self-assembly was determined. Subsequently, the Doxy loading efficiency of DA3 was assessed. The cytotoxicity of Doxy, DA3, PEI, and DA-Doxy in nasal fibroblasts was evaluated using the WST-1 assay. The anti-tissue remodeling and anti-inflammatory effects of DA3-Doxy and DA3 were examined using real-time polymerase chain reaction (Real-time PCR), immunocytochemistry, western blot, and Sircol assay. RESULTS: Both DA3 and DA3-Doxy exhibited cytotoxicity at 10 µg/ml in nasal fibroblasts. Doxy partially inhibited α-smooth muscle actin, collagen types I and III, and fibronectin. However, DA3-Doxy significantly inhibited α-SMA, collagen types I and III, and fibronectin at 5 µg/ml. DA3-Doxy also modulated TGF-ß1-induced changes in the expression of MMP 1, 2, and 9. Nonetheless, TGF-ß1-induced expression of MMP3 was further increased by DA3-Doxy. The expression of TIMP 1 and 2 was partially reduced with 5 µg/ml DA3-Doxy. CONCLUSIONS: Although initially developed for the delivery of genetic materials or drugs, DA3 exhibits inhibitory effects on myofibroblast differentiation and ECM production. Therefore, it holds therapeutic potential for CRS, and a synergistic effect can be expected when loaded with CRS treatment drugs.
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Diferenciação Celular , Ácido Desoxicólico , Doxiciclina , Fibroblastos , Polietilenoimina , Humanos , Polietilenoimina/química , Polietilenoimina/farmacologia , Ácido Desoxicólico/química , Ácido Desoxicólico/farmacologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Diferenciação Celular/efeitos dos fármacos , Doxiciclina/farmacologia , Doxiciclina/química , Matriz Extracelular/metabolismo , Matriz Extracelular/efeitos dos fármacos , Fator de Crescimento Transformador beta1/metabolismo , Miofibroblastos/efeitos dos fármacos , Miofibroblastos/metabolismo , Mucosa Nasal/efeitos dos fármacos , Mucosa Nasal/metabolismo , Mucosa Nasal/citologia , Actinas/metabolismoRESUMO
Allergic rhinitis is a chronic nasal mucosal inflammation characterized by upper airway hyperresponsiveness, involving a variety of immune cells and inflammatory mediators. Drugs, immunotherapy, and surgical operation are the principal treatments at present. The study found that mesenchymal stem cells have the ability of immune regulation and have a promising clinical application in the treatment of allergic rhinitis. In this review, the action mechanism of mesenchymal stem cells, the immunomodulatory effect of mesenchymal stem cells on the key cells of allergic rhinitis, and the challenges of clinical application are reviewed, to provide new directions for the treatment of allergic rhinitis.
Assuntos
Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Mucosa Nasal , Rinite Alérgica , Humanos , Células-Tronco Mesenquimais/citologia , Rinite Alérgica/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Mucosa Nasal/citologiaRESUMO
BACKGROUND: The role of Akt in nasal immunity is unstudied. Akt phosphorylates and activates endothelial nitric oxide synthase (eNOS) expressed in epithelial ciliated cells. Nitric oxide (NO) production by ciliated cells can have antibacterial and antiviral effects. Increasing nasal NO may be a useful antipathogen strategy in chronic rhinosinusitis (CRS). We previously showed that small-molecule Akt activator SC79 induces nasal cell NO production and suppresses IL-8 via the transcription factor Nrf-2. We hypothesized that SC79 NO production may additionally have antibacterial effects. METHODS: NO production was measured using fluorescent dye DAF-FM. We tested effects of SC79 during co-culture of Pseudomonas aeruginosa with primary nasal epithelial cells, using CFU counting and live-dead staining to quantify bacterial killing. Pharmacology determined the mechanism of SC79-induced NO production and tested dependence on Akt. RESULTS: SC79 induced dose-dependent, Akt-dependent NO production in nasal epithelial cells. The NO production required eNOS and Akt. The NO released into the airway surface liquid killed P. aeruginosa. No toxicity (LDH release) or inflammatory effects (IL8 transcription) were observed over 24 h. CONCLUSIONS: Together, these data suggest multiple immune pathways are stimulated by SC79, with antipathogen effects. This in vitro pilot study suggests that a small-molecule Akt activator may have clinical utility in CRS or respiratory other infection settings, warranting future in vivo studies.
Assuntos
Células Epiteliais , Mucosa Nasal , Óxido Nítrico , Proteínas Proto-Oncogênicas c-akt , Pseudomonas aeruginosa , Humanos , Óxido Nítrico/metabolismo , Mucosa Nasal/imunologia , Mucosa Nasal/metabolismo , Mucosa Nasal/citologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/imunologia , Células Cultivadas , Óxido Nítrico Sintase Tipo III/metabolismo , Antibacterianos/farmacologia , Rinite/imunologia , Rinite/microbiologia , Rinite/tratamento farmacológicoRESUMO
Influenza B viruses (IBV) cocirculate with influenza A viruses (IAV) and cause periodic epidemics of disease, yet antibody and cellular responses following IBV infection are less well understood. Using the ferret model for antisera generation for influenza surveillance purposes, IAV resulted in robust antibody responses following infection, whereas IBV required an additional booster dose, over 85% of the time, to generate equivalent antibody titers. In this study, we utilized primary differentiated ferret nasal epithelial cells (FNECs) which were inoculated with IAV and IBV to study differences in innate immune responses which may result in differences in adaptive immune responses in the host. FNECs were inoculated with IAV (H1N1pdm09 and H3N2 subtypes) or IBV (B/Victoria and B/Yamagata lineages) and assessed for 72 h. Cells were analyzed for gene expression by quantitative real-time PCR, and apical and basolateral supernatants were assessed for virus kinetics and interferon (IFN), respectively. Similar virus kinetics were observed with IAV and IBV in FNECs. A comparison of gene expression and protein secretion profiles demonstrated that IBV-inoculated FNEC expressed delayed type-I/II IFN responses and reduced type-III IFN secretion compared to IAV-inoculated cells. Concurrently, gene expression of Thymic Stromal Lymphopoietin (TSLP), a type-III IFN-induced gene that enhances adaptive immune responses, was significantly downregulated in IBV-inoculated FNECs. Significant differences in other proinflammatory and adaptive genes were suppressed and delayed following IBV inoculation. Following IBV infection, ex vivo cell cultures derived from the ferret upper respiratory tract exhibited reduced and delayed innate responses which may contribute to reduced antibody responses in vivo.IMPORTANCEInfluenza B viruses (IBV) represent nearly one-quarter of all human influenza cases and are responsible for significant clinical and socioeconomic impacts but do not pose the same pandemic risks as influenza A viruses (IAV) and have thus received much less attention. IBV accounts for greater severity and deaths in children, and vaccine efficacy remains low. The ferret can be readily infected with human clinical isolates and demonstrates a similar course of disease and immune responses. IBV, however, generates lower antibodies in ferrets than IAV following the challenge. To determine whether differences in initial innate responses following infection may affect the development of robust adaptive immune responses, ferret respiratory tract cells were isolated, infected with IAV/IBV, and compared. Understanding the differences in the initial innate immune responses to IAV and IBV may be important in the development of more effective vaccines and interventions to generate more robust protective immune responses.
Assuntos
Imunidade Adaptativa , Células Epiteliais , Furões , Imunidade Inata , Vírus da Influenza A , Vírus da Influenza B , Interferons , Mucosa Nasal , Animais , Criança , Humanos , Anticorpos Antivirais/análise , Anticorpos Antivirais/biossíntese , Anticorpos Antivirais/imunologia , Modelos Animais de Doenças , Células Epiteliais/citologia , Células Epiteliais/imunologia , Células Epiteliais/virologia , Furões/imunologia , Furões/virologia , Vírus da Influenza A/classificação , Vírus da Influenza A/crescimento & desenvolvimento , Vírus da Influenza A/imunologia , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Vírus da Influenza B/classificação , Vírus da Influenza B/crescimento & desenvolvimento , Vírus da Influenza B/imunologia , Vacinas contra Influenza , Influenza Humana/virologia , Interferons/imunologia , Mucosa Nasal/citologia , Mucosa Nasal/imunologia , Mucosa Nasal/virologia , Linfopoietina do Estroma do Timo/genética , Linfopoietina do Estroma do Timo/imunologia , Células CultivadasRESUMO
INTRODUCCIÓN: La citología nasal es utilizada como método complementario diagnóstico de la rinitis y el hallazgo de eosinófilos; en la misma aumenta la sensibilidad para confirmar una alergia a casi el 80%. Los corticoesteroides intranasales tópicos se utilizan actualmente como primera línea de tratamiento de la rinitis porque reducen la inflamación de la mucosa que subyace a los signos y síntomas de la enfermedad. MATERIAL Y MÉTODOS: Estudio observacional, retrospectivo y analítico. Se incluyeron pacientes entre 16 a 65 años edad inclusive, que consultaron al Servicio de Otorrinolaringología y Alergia e Inmunología de la Clínica Universitaria Reina Fabiola entre agosto de 2016 y julio de 2017 con diagnóstico de rinitis alérgica. La información de las variables cuantitativas se sometió a una comprobación estadística realizada mediante el test de Wilcoxon apareado. Se consideró significativo un valor de p<0,05
INTRODUCTION: Nasal cytology is used as a complementary diagnostic method of rhinitis and the finding of eosinophils in it increases the sensitivity to confirm an allergy to almost 80%. Topical intranasal corticosteroids are currently used as the first line of treatment for rhinitis because they reduce the inflammation of the mucosa that underlies the signs and symptoms of the disease. MATERIAL AND METHODS: Observational, retrospective and analytical study. Patients between the ages of 16 and 65 years were included, who consulted the Otorhinolaryngology and Allergy and Immunology Department of the Reina Fabiola University Clinic between August 2016 and July 2017 with a diagnosis of allergic rhinitis. The information of the quantitative variables was subjected to a statistical check carried out by means of the paired Wilcoxon test. A value of p...
Assuntos
Adolescente , Adulto , Corticosteroides/uso terapêutico , Eosinófilos/efeitos dos fármacos , Rinite Alérgica/imunologia , Mucosa Nasal/citologia , Propionatos/uso terapêutico , Estudos Retrospectivos , Citodiagnóstico/estatística & dados numéricos , Estudo Observacional , Rinite Alérgica/tratamento farmacológico , Furoato de Mometasona/uso terapêuticoRESUMO
Abstract Introduction: In teenagers with perennial allergic rhinitis, exposure to tobacco cigarette smoke increases the count of eosinophils in the nasal mucosa; the recruitment of eosinophils arises from the combined action of a number of cellular and molecular signals, including eotaxin. Objective: To assess the effect of exposure to tobacco cigarette smoke on the count of immunoreactive cells to eotaxin-1 and eosinophils on the nasal mucosa of children and teenagers with perennial allergic rhinitis. Methods: In a cross-sectional study, forty-four patients were evaluated (aged 7-19 years old): 22 with and 22 with no exposure to tobacco cigarette smoke. After replying to 2 validated questionnaires, on Asthma and Allergies in Childhood and on the severity of nasal symptoms, nasal mucosal samples were obtained by scraping the middle one-third of the inferior turbinates. Then counts of immunoreactive cells to eotaxin-1 and eosinophils were assessed by immunohistochemistry. Results: Patients with exposure to tobacco cigarette smoke showed higher cell counts of both eotaxin-1 and eosinophils than patients with no exposure to the smoke, with no correlation between the two variables. However, both counts, of eotaxin-1 and eosinophils, were related to the cotinine/creatinine ratio. Conclusions: Exposure to tobacco cigarette smoke can increase eotaxin-1 and the count of eosinophils in the nasal mucosa of young patients with perennial allergic rhinitis.
Resumo Introdução: Em adolescentes com rinite alérgica perene, a exposição à fumaça do cigarro de tabaco aumenta a contagem de eosinófilos na mucosa nasal. O recrutamento de eosinófilos surge da ação combinada de alguns sinais celulares e moleculares, inclusive a eotaxina. Objetivo: Avaliar o efeito da exposição à fumaça do cigarro de tabaco na contagem de células imunorreativas a eotaxina-1 e eosinófilos na mucosa nasal de crianças e adolescentes com rinite alérgica perene. Método: Em um estudo transversal, 44 pacientes foram avaliados (entre sete e 19 anos): 22 com e 22 sem exposição à fumaça do cigarro de tabaco. Depois de responder a dois questionários validados, sobre asma e alergias na infância e sobre a gravidade dos sintomas nasais, as amostras de mucosa nasal foram obtidas por meio de raspagem do terço médio das conchas inferiores. Em seguida, as contagens de células imunorreativas para eotaxina-1 e eosinófilos foram avaliadas por imuno-histoquímica. Resultados: Os pacientes com exposição à fumaça do cigarro de tabaco apresentaram contagens de células mais elevadas tanto para eotaxina-1 como para eosinófilos em comparação com os pacientes sem exposição à fumaça, sem correlação entre as duas variáveis. No entanto, ambas as contagens, de eotaxina-1 e eosinófilos foram relacionadas com a razão cotinina/creatinina. Conclusões: A exposição à fumaça do cigarro de tabaco pode aumentar a eotaxina-1 e a contagem de eosinófilos na mucosa nasal de pacientes jovens com rinite alérgica perene.
Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Adulto , Adulto Jovem , Poluição por Fumaça de Tabaco/efeitos adversos , Rinite Alérgica Sazonal/imunologia , Eosinófilos/imunologia , Quimiocina CCL11/imunologia , Mucosa Nasal/imunologia , Índice de Gravidade de Doença , Imuno-Histoquímica , Contagem de Células , Rinite Alérgica Sazonal/patologia , Estudos Transversais , Eosinófilos/citologia , Quimiocina CCL11/análise , Mucosa Nasal/citologia , Mucosa Nasal/químicaRESUMO
The respiratory epithelium is the first line of contact with the external hazards. Thus it can be damaged and need to be replaced to avoid healing by fibrosis. Tracheal tissue engineering is an alternative promising treatment modality. Mesenchymal stem cell markers are surface proteins, which are responsible for some of these cells unique properties. The objective of this study was to detect the mesenchymal stem cell phenotype among the human nasal respiratory epithelial cells via two immunophenotyping techniques. Respiratory epithelial cells were cultured using co-culture technique, fibroblasts was removed at confluence leaving respiratory epithelial cells, which were passage further to passage 4. Cells were evaluated for mesenchymal stem cell markers that were CD73, CD90, CD105 and the hematopoietic stem cell marker CD45 at passage 1 (P1) and passage 4 (P4) using Flow cytometry and Immunocytochemistry techniques. Respiratory epithelial cells expressed the mesenchymal stem cell markers at P1 and maintain the expression these markers until P4. Using both techniques, to compare the values of mesenchymal stem cell markers expression at P1 to P4 there was no significant difference. This study indicates that respiratory epithelial cells derived from nasal turbinate retain some of mesenchymal stem cells properties even after serial passages. Both methods of Immunophenotyping are comparable.
El epitelio respiratorio es la primera línea de contacto con los peligros externos. Por lo tanto, puede ser dañado y necesita ser reemplazado para evitar uan cicatrización por fibrosis. La ingeniería de tejidos traqueales es una modalidad de tratamiento alternativo prometedora. Los marcadores de células troncales mesenquimales son proteínas de superficie, que son responsables de algunas propiedades únicas de estas células. El objetivo fue detectar el fenotipo de células troncales mesenquimales entre las células epiteliales respiratorias nasales humanas a través de dos técnicas de inmunofenotipaje. Fueron cultivadas las células epiteliales respiratorias utilizando la técnica de co-cultivo; los fibroblastos se eliminaron en la confluencia dejando solo células epiteliales respiratorias, resultantes de los 4 pasajes. Las células fueron evaluadas para encontrar marcadores de células troncales mesenquimales mediante CD73, CD90, CD105 y el marcador de células troncales hematopoyéticas CD45 en el paso 1 (P1) y el paso 4 (P4), usando citometría de flujo y técnicas de inmunocitoquímica. Las células epiteliales respiratorias expresaron los marcadores de células troncales mesenquimales en P1 y mantuvieron la expresión de estos marcadores hasta P4. No hubo diferencias significativas en el uso de ambas técnicas al comparar los valores de los marcadores de células troncales mesenquimales expresadas desde P1 a P4. Este estudio indica que las células epiteliales respiratorias derivadas de la concha nasal retienen algunas de las propiedades de células troncales mesenquimales, incluso después de pases seriados. Ambos métodos de inmunofenotipificación son comparables.
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Humanos , Biomarcadores/metabolismo , Células Epiteliais/citologia , Mucosa Nasal/citologia , Conchas Nasais/citologia , Técnicas de Cultura de Células , Citometria de Fluxo , Imuno-Histoquímica , Células-Tronco Mesenquimais/citologia , Fenótipo , Engenharia TecidualRESUMO
OBJECTIVES: This study intends to provide a quick, easy, and inexpensive way to assess nuclear abnormalities such as micronuclei and bud frequencies; binucleated, karyorrhectic, karyolytic, pycnotic, and condensed chromatin cells in nasal scrapings of infants, which are particularly important for conducting genotoxic studies related to the inhaled atmosphere in pediatric populations. METHODS: Nasal swab samples were collected from 40 infants under 12 months of age using a small cytobrush. 2,000 cells from each infant sample were analyzed and classified according to the frequency of nuclear abnormalities. RESULTS: Rates of nuclear abnormalities found agree with values reported in other studies of neonates and children. This study found 0.13% of cells with micronuclei; 1.20% karyorrhexis; 0.03% pyknosis; 10.85% karyolysis; 1.11% condensed chromatin; 0.54 binucleated cells; and 0.02% nuclear bud. Differences were not observed between genders or environmental passive smoking, nor was any age correlation found. CONCLUSION: The assay proposed here is suitable for assessing the frequency of nuclear abnormalities from nasal cells in infants. .
OBJETIVOS: Este estudo pretendeu fornecer uma forma rápida, fácil e barata de avaliar anormalidades nucleares, como frequências de micronúcleos e gêmea, células binucleadas, cariorréticas, cariolíticas, picnóticas e com cromatina condensada, em esfregados nasais de neonatos, o que é particularmente importante para a realização de estudos genotóxicos relacionados ao ar inalado nas populações pediátricas. MÉTODOS: Foram coletadas amostras de esfregaço nasal de 40 neonatos com menos de 12 meses de idade, utilizando uma pequena escova citológica. Foram analisadas 2.000 células da amostra de cada neonato e classificadas de acordo com a frequência de anormalidades nucleares. RESULTADOS: As taxas de anormalidades nucleares encontradas neste estudo são compatíveis com os valores relatados em outros estudos de neonatos e crianças. Encontramos 0,13% de células com micronúcleos, 1,20% com cariorrexe, 0,03% com picnose, 10,85% com cariólise, 1,11% com cromatina condensada, 0,54 com células binucleadas e 0,02% com células nucleares gêmeas. Não observamos diferenças entre os gêneros, tabagismo passivo e nenhuma correlação entre idades. CONCLUSÃO: O ensaio proposto neste estudo é adequado para avaliar a frequência de anormalidades nucleares de células nasais em neonatos. .
Assuntos
Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Poluição do Ar/efeitos adversos , Núcleo Celular/genética , Dano ao DNA , Mucosa Nasal/citologia , Testes para Micronúcleos/métodos , Poluição por Fumaça de Tabaco/efeitos adversosRESUMO
Determinar y comparar las características citológicas de los epitelios nasal, faríngeo y vaginal en mujeres adultas jóvenes. Estudio prospectivo y transversal de 35 mujeres no embarazadas, entre 18 y 35 años de edad, durante el período comprendido entre el 01 de enero al 01 de julio de 2004; que ingirieron o no anticonceptivos orales y a quienes se les estudiaron las citologías de los epitelios vaginal, faríngeo y nasal, según el porcentaje de índice de maduración de células exfoliadas y la cuantificación sérica de estrógeno y progesterona. En el Departamento de Obstetricia y Ginecología Hospital Chiquinquirá de Maracaibo, Estado Zulia. Se determinó que no existe diferencia significativa entre la celularidad de los epitelios estudiados. Se estableció que existe buena correlación entre las células superficiales e intermedias de los frotis de vagina, faringe y nariz durante las fases del ciclo menstrual, en todas las mujeres, aun en las que recibieron píldoras anticonceptivas. Los niveles de estrógenos y progesterona tuvieron concordancia con las fases del ciclo, con el porcentaje y tipo de células exfoliadas. Los resultados obtenidos demuestran que los epitelios nasales y faríngeos responden al influjo de hormonas ováricas similarmente como ocurre en la vagina y que estos métodos pueden ser aplicados en condiciones de difícil acceso a laboratorios hormonales o en pacientes vírgenes o con atresia/agenesia vaginal y niñas
To determine and compare the cytological characteristics of nasal, pharyngeal and vaginal epitheliums in young adult women. A prospective and transversal study of 35 non pregnant women of 18 to 35 years old during the period January 1, 2004 - November 2004; some on contraceptives pills, were studied by analysis of smear of nasal, pharyngeal and vaginal trough maturation index of exfoliated cells and serum level of estrogen and progesterone. Department of Obstetrics and Gynecology, Hospital Chiquinquira, Maracaibo, Estado Zulia. Venezuela. It was established that exists a good correlation between the superficial and intermediate cells in the different phases of the menstrual cycle and those who received oral contraceptives and the group that did not. Is established that there are correlation between the cells when compared vaginal-pharynx and vaginal-nose, in both groups. The serum levels of estrogen and progesterone were accorded to the cycle phases and with the percentage of cells in both, medicated and non medicated patients. Ours results demonstrate that the nasal and pharyngeal epitheliums respond to ovarian hormones, similar to what occurs in the vagina. It is recommended to use this method in those patients where it is need to investigate the endocrine status and where it is difficult to reach the vagina: nubile girls, vaginal agenesis, imperforate hymen; or where there is not a endocrine laboratory near
Assuntos
Feminino , Anticoncepcionais Orais/administração & dosagem , Anticoncepcionais Orais/efeitos adversos , Ciclo Menstrual/fisiologia , Células Epiteliais/citologia , Mucosa Laríngea/citologia , Mucosa Nasal/citologia , Técnicas Citológicas/métodos , Vagina/citologia , Citodiagnóstico , Estrogênios/análise , Progesterona/análiseRESUMO
Introdução: Embora recomendado, o estudo da citologia nasal não é realizado rotineiramente devido à falta de padronização, realização de contagens somente qualitativas, e a escassez de material obtido. Objetivos: Nosso estudo avaliou a possibilidade de realização de uma leitura quantitativa de esfregaços nasais por cotonete e por escova em pacientes com rinite, e comparou os achados obtidos através das duas técnicas. Casuística e Métodos: Foram selecionados sessenta pacientes apresentando história clínica de rinite. Todos foram submetidos, em uma mesma avaliação, sequencialmente, a um esfregaço por cotonete na narina esquerda e a um por escova na direita. O esfregaço foi realizado em lâmina de vidro, com análise qualitativa e quantitativa de eosinófilos, neutrófilos e células epiteliais. Os achados foram comparados através do coeficiente de correlação intraclasse e do coeficiente Kappa, quando apropriado. Resultados: Observamos que a análise quantitativa dos esfregaços em lâmina foi possível, porém dificultada por questões associadas à técnica do exame. Qualitativamente não houve diferença de achado dos três tipos celulares entre as duas técnicas, mas sim quantitativo e de forma significativa, sendo maior para o escovado. Conclusões: Concluímos que ambos os procedimentos são de fácil execução, baixo custo operacional, viáveis para realização ambulatorial, e de boa tolerância.
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Citodiagnóstico , Mucosa Nasal/citologia , Rinite/imunologia , Contagem de Células , Estudos Transversais , Células Epiteliais/imunologia , Eosinófilos/imunologia , Neutrófilos/imunologia , Projetos PilotoRESUMO
Con el objeto de determinar si la citología nasal en niños con rinitis crónica tiene relación con sinusitis paranasal, se hizo un estudio prospectivo, descriptivo y comparativo en 48 niños entre 9 meses y 9 años, de ambos sexos, de septiembre de 1994 a agosto de 1996 por presentar datos clínicos compatibles con rinitis crónica. El grupo control fue de pacientes encamados en las salas de pediatría por padecimiento no respiratorios, hematológicos o inflamatorios. Se tomó citología exfoliativa de la mucosa nasal del cornete inferior en ambas fosas y se relacionaron los hallazgos considerados como inflamación crónica de la nariz con opacidad de los senos paranasales en radiografías simples. Se encontró relación significativa (p<0.05) entre la presencia de metaplasia y neutrófilos. Se concluye que la célula de metaplasia y neutrófilos. Se concluye que la célula de metaplasia escamosa puede ser una célula adaptada o en respuesta a la inflamación crónica de la mucosa nasal donde el neutrófilo es la célula inflamatoria predominante. La citología nasal con presencia de metaplasia escamosa y neutrófilos es un probable predictor de sinusitis
Assuntos
Humanos , Masculino , Feminino , Seios Paranasais/citologia , Seios Paranasais/fisiopatologia , Seios Paranasais , Sinusite/diagnóstico , Rinite/diagnóstico , Estudos Prospectivos , Biologia Celular , Mucosa Nasal/citologia , Sinais e Sintomas RespiratóriosRESUMO
Antecedentes: desde hace más de una centuria el eosinófilo se ha considerado decisivo en la producción de asma. Material y método: se estudiaron 600 sujetos, elegidos al azar, con edades entre 1 y 55 años, 300 son asmáticos y el resto estaban aparentemente sanos. Resultados: en los 300 asmáticos, la concentración media de eosinófilos en sangre periférica fue de 615.93/m al cubo, (p=0.000004, R = 2.13), los asmáticos con concentraciónes de eosinófilos circulantes superiores a 400/mm al cubo tuvieron más cuadros de asma agudizada por mes (p= NS) y demandaron con mayor frecuencia los servicios hospitalarios para su estabilización (p = 0.00001, R = 3.56). En 195 enfermos se detectaron eosinófilos en el moco nasal con una concentración media de 10.54 por ciento. (p= 0.04, R= 0.70). Conclusiones: la incidencia y prevalencia del asma se ve favorecida por la carencia de sensores predictivos. En este estudio los eosinófilos sanguíneos y nasales en niveles elevados se identificaron en sujetos de alto riesgo; en consecuencia, se propone su utilización para el diagnóstico oportuno y seguimiento a fin de aplicar medidas en la fase de equilibrio de la cadena salud-enfermedad
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Adolescente , Adulto , Pessoa de Meia-Idade , Asma/parasitologia , Asma/patologia , Eosinofilia/patologia , Eosinófilos , Enteropatias Parasitárias/diagnóstico , Enteropatias Parasitárias/parasitologia , Muco/citologia , Mucosa Nasal/citologiaRESUMO
Durante el período de enero de 1992 a febrero de 1996 se estudiaron 454 citologías nasales y faríngeas en cuadros inflamatorios infecciosos. La Chlamydia trachomatis (CT) fue el agente etiólogico en 183 (67.80 por ciento) de los frotis de las fosas nasales, mientras que en 271 frotis faríngeos, representó el 8.11 por ciento (22 casos). De 124 pacientes con infección por Chlamydia trachomatis en las fosas nasales, en 15 (12.09 por ciento) se identificó Chlamydia en la faringe; en 18 pacientes (14.51 por ciento) se identificó conjuntivitis por CT y en 3 pacientes de los 4 en los que fue posible tomar muestras conjuntivales, nasales y fáringeas se encontró a la CT
Assuntos
Infecções por Chlamydia/diagnóstico , Infecções por Chlamydia/tratamento farmacológico , Infecções por Chlamydia/imunologia , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/efeitos dos fármacos , Chlamydia trachomatis/imunologia , Chlamydia trachomatis/isolamento & purificação , Mucosa Nasal/citologia , Mucosa Nasal/microbiologia , Faringe/citologia , Faringe/microbiologia , Rinite/etiologia , Camada de EsfregaçoRESUMO
El objetivo del estudio fue investigar la citopatología nasal en pacientes fumadores, fumadores pasivos y no fumadores. Se estudiaron 60 pacientes divididos en grupos de 20 cada uno. En el 95 por ciento de los fumadores (Grupo 1) se encontró algún tipo de metaplasia o displasia; en los fumadores pasivos (Grupo 2) solo en el 55 por ciento se encontró alteración citológica y solo en el 15 por ciento de los no fumadores (Grupo 3). El estudio concluye que el riesgo de tener cambios citológicos en los fumadores pasivos es de aproximadamente la mitad en relación a los fumadores. La citología nasal es un método barato, sencillo y muy adecuado para el diagnóstico de los cambios histológicos de la mucosa nasal