RESUMEN
Innate lymphoid cells (ILCs) are the most recently described group of lymphoid subpopulations. These tissue-resident cells display a heterogeneity resembling that observed on different groups of T cells, hence their categorization as cytotoxic NK cells and helper ILCs type 1, 2 and 3. Each one of these groups is highly diverse and expresses different markers in a context-dependent manner. Type 2 innate lymphoid cells (ILC2s) are activated in response to helminth parasites and regulate the immune response. They are involved in the etiology of diseases associated with allergic responses as well as in the maintenance of tissue homeostasis. Markers associated with their identification differ depending on the tissue and model used, making the study and understanding of these cells a cumbersome task. This review compiles evidence for the heterogeneity of ILC2s as well as discussion and analyses of molecular markers associated with their identity, function, tissue-dependent expression, and how these markers contribute to the interaction of ILC2s with specific microenvironments to maintain homeostasis or respond to pathogenic challenges.
Asunto(s)
Antígenos de Diferenciación/análisis , Subgrupos Linfocitarios/inmunología , Tejido Adiposo Blanco/inmunología , Tejido Adiposo Blanco/patología , Animales , Citocinas/metabolismo , Helmintiasis/inmunología , Antígenos de Histocompatibilidad Clase II/inmunología , Homeostasis , Humanos , Inmunofenotipificación , Inflamación , Intestinos/inmunología , Pulmón/inmunología , Subgrupos Linfocitarios/química , Ratones , Nutrientes , Especificidad de Órganos , Proteínas Proto-Oncogénicas c-kit/inmunología , Receptores de Superficie Celular/inmunología , Piel/inmunología , Factor de Células Madre/inmunologíaRESUMEN
INTRODUCTION: The olfactory neuro-epithelium has an intrinsic capability of renewal during lifetime provided by the existence of globose and horizontal olfactory precursor cells. Additionally, mesenchymal stromal olfactory cells also support the homeostasis of the olfactory mucosa cell population. Under in vitro culture conditions with Dulbecco modified eagle/F12 medium supplemented with 10% fetal bovine serum, tissue biopsies from upper turbinate have generated an adherent population of cells expressing mainly mesenchymal stromal phenotypic markers. A closer examination of these cells has also found co-expression of olfactory precursors and ensheathing cell phenotypic markers. These results were suggestive of a unique property of olfactory mesenchymal stromal cells as potentially olfactory progenitor cells. OBJECTIVE: To study whether the expression of these proteins in mesenchymal stromal cells is modulated upon neuronal differentiation. MATERIALS AND METHODS: We observed the phenotype of olfactory stromal cells under DMEM/F12 plus 10% fetal bovine serum in comparison to cells from spheres induced by serum-free medium plus growth factors inducers of neural progenitors. RESULTS: The expression of mesenchymal stromal (CD29+, CD73+, CD90+, CD45-), horizontal basal (ICAM-1/CD54+, p63+, p75NGFr+), and ensheathing progenitor cell (nestin+, GFAP+) proteins was determined in the cultured population by flow cytometry. The determination of Oct 3/4, Sox-2, and Mash-1 transcription factors, as well as the neurotrophins BDNF, NT3, and NT4 by RT-PCR in cells, was indicative of functional heterogeneity of the olfactory mucosa tissue sample. CONCLUSIONS: Mesenchymal and olfactory precursor proteins were downregulated by serum-free medium and promoted differentiation of mesenchymal stromal cells into neurons and astroglial cells.
Introducción. El recambio celular del neuroepitelio olfatorio ocurre durante la vida del individuo gracias a precursores olfatorios. Además, las células mesenquimales del estroma también contribuyen a la homeostasis de la mucosa. Cuando un explante de una biopsia de mucosa se cultiva en un medio esencial mínimo, se genera una población predominante de células adherentes que expresan proteínas típicas de las células mesenquimales del estroma. La coexpresión de marcadores fenotípicos de precursores olfatorios y de células del recubrimiento del nervio olfatorio constituiría una propiedad única de las células mesenquimales del estroma. Objetivo. Determinar si la diferenciación celular de las células mesenquimales hacia fenotipos neurales modula la expresión de los marcadores mesenquimales característicos. Materiales y métodos. Se compararon las células aisladas de la mucosa olfatoria en un medio de cultivo con suplemento de 10 % de suero fetal bovino con esferas generadas en un medio sin suero más factores de crecimiento. Resultados. Se determinó la expresión de proteínas de las células mesenquimales del estroma (CD29+, CD73+, CD90+, CD45-), de las basales horizontales (ICAM-1/CD54+, p63+, p75NGFr+), y de las del recubrimiento del nervio olfatorio (nestin+, GFAP+) en la misma población cultivada. La determinación de Oct 3/4, Sox-2 y Mash-1, así como de las neurotrofinas BDNF, NT3 y NT4, sugirió que las células del estroma son funcionales. La expresión de las proteínas de las células mesenquimales y los precursores olfatorios, disminuyó en las células de las mesenesferas inducidas por ausencia de suero en el medio de cultivo. Conclusión. Las células mesenquimales del estroma de la mucosa olfatoria presentan una tendencia dominante hacia la diferenciación neural.
Asunto(s)
Células Madre Mesenquimatosas/metabolismo , Mucosa Nasal/citología , Mucosa Olfatoria/citología , Biosíntesis de Proteínas , Adipogénesis , Antígenos de Diferenciación/análisis , Técnicas de Cultivo de Célula , Diferenciación Celular , Células Cultivadas , Condrogénesis , Medios de Cultivo/farmacología , Medio de Cultivo Libre de Suero/farmacología , Proteína Ácida Fibrilar de la Glía/biosíntesis , Proteína Ácida Fibrilar de la Glía/genética , Humanos , Péptidos y Proteínas de Señalización Intercelular/farmacología , Células Madre Mesenquimatosas/efectos de los fármacos , Mucosa Nasal/metabolismo , Factores de Crecimiento Nervioso/biosíntesis , Factores de Crecimiento Nervioso/genética , Nestina/biosíntesis , Nestina/genética , Neuroglía/metabolismo , Neuronas/metabolismo , Mucosa Olfatoria/metabolismo , Osteogénesis , Proteínas Recombinantes/farmacología , Esferoides Celulares , Factores de Transcripción/biosíntesis , Factores de Transcripción/genética , Cornetes NasalesRESUMEN
INTRODUCTION: Hypersensitivity, local irritative and cytotoxic effects are known for the chemical components of Syzygium aromaticum and Cinnamomum zeylanicum contained in dental materials. However, there is no intimate data in dentistry using the whole extracts of these plants and introducing new ones. Salvia triloba is a well-known anti-inflammatory plant that correspondingly could be used in several dental traumas. OBJECTIVES: We aimed to show and compare the effect of S. aromaticum, C. zeylanicum, and S. triloba extracts on dental pulp stem cells (DPSCs) proliferation, differentiation, and immune responses. MATERIAL AND METHODS: Using xCELLigence, a real time monitoring system, we obtained a growth curve of DPSCs with different concentrations of the Extracts. A dose of 10 µg/mL was the most efficient concentration for vitality. Osteogenic differentiation and anti-inflammatory activities were determined by using an ELISA Kit to detect early and late markers of differentiation. RESULTS: The level of osteonectin (ON, early osteogenic marker) decreased, which indicated that the osteogenic differentiation may be accelerated with addition of extracts. However, the level of osteocalcin (OCN, late osteogenic marker and sign of calcium granulation) differed among the extracts, in which S. aromaticum presented the highest value, followed by S. triloba and C. zeylanicum. Surprisingly, the determined calcium granules were reduced in S. aromaticum and S. triloba. In response to tumor necrosis factor alpha (TNF-α), S. triloba-treated DPSCs showed the most reduced level of IL-6 cytokine level. We suggest C. zeylanicum as a promising osteogenic inducer and S. triloba as a potent anti-inflammatory agent, which could be used safely in biocomposite or scaffold fabrications for dentistry. CONCLUSIONS: Because calcium granule formation and cell viability play a critical role in hard tissue formation, S. aromaticum in dentistry should be strictly controlled, and the mechanism leading to reduced calcium granule formation should be identified.
Asunto(s)
Antiinflamatorios/farmacología , Cinnamomum zeylanicum/química , Pulpa Dental/citología , Medicamentos Herbarios Chinos/farmacología , Células Madre Mesenquimatosas/efectos de los fármacos , Extractos Vegetales/farmacología , Syzygium/química , Adolescente , Análisis de Varianza , Antígenos de Diferenciación/análisis , Calcio/análisis , Canfanos , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Citocinas/análisis , Pulpa Dental/efectos de los fármacos , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Humanos , Osteocalcina/análisis , Osteogénesis/efectos de los fármacos , Osteonectina/análisis , Panax notoginseng , Reproducibilidad de los Resultados , Salvia miltiorrhiza , Factores de Tiempo , Adulto JovenRESUMEN
Abstract Hypersensitivity, local irritative and cytotoxic effects are known for the chemical components of Syzygium aromaticum and Cinnamomum zeylanicum contained in dental materials. However, there is no intimate data in dentistry using the whole extracts of these plants and introducing new ones. Salvia triloba is a well-known anti-inflammatory plant that correspondingly could be used in several dental traumas. Objectives: We aimed to show and compare the effect of S. aromaticum, C. zeylanicum, and S. triloba extracts on dental pulp stem cells (DPSCs) proliferation, differentiation, and immune responses. Material and Methods: Using xCELLigence, a real time monitoring system, we obtained a growth curve of DPSCs with different concentrations of the Extracts. A dose of 10 μg/mL was the most efficient concentration for vitality. Osteogenic differentiation and anti-inflammatory activities were determined by using an ELISA Kit to detect early and late markers of differentiation. Results: The level of osteonectin (ON, early osteogenic marker) decreased, which indicated that the osteogenic differentiation may be accelerated with addition of extracts. However, the level of osteocalcin (OCN, late osteogenic marker and sign of calcium granulation) differed among the extracts, in which S. aromaticum presented the highest value, followed by S. triloba and C. zeylanicum. Surprisingly, the determined calcium granules were reduced in S. aromaticum and S. triloba. In response to tumor necrosis factor alpha (TNF-α), S. triloba-treated DPSCs showed the most reduced level of IL-6 cytokine level. We suggest C. zeylanicum as a promising osteogenic inducer and S. triloba as a potent anti-inflammatory agent, which could be used safely in biocomposite or scaffold fabrications for dentistry. Conclusions: Because calcium granule formation and cell viability play a critical role in hard tissue formation, S. aromaticum in dentistry should be strictly controlled, and the mechanism leading to reduced calcium granule formation should be identified.
Asunto(s)
Humanos , Adolescente , Adulto Joven , Medicamentos Herbarios Chinos/farmacología , Extractos Vegetales/farmacología , Cinnamomum zeylanicum/química , Syzygium/química , Pulpa Dental/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Antiinflamatorios/farmacología , Osteogénesis/efectos de los fármacos , Factores de Tiempo , Ensayo de Inmunoadsorción Enzimática , Antígenos de Diferenciación/análisis , Osteocalcina/análisis , Osteonectina/análisis , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Calcio/análisis , Reproducibilidad de los Resultados , Análisis de Varianza , Citocinas/análisis , Pulpa Dental/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Citometría de FlujoRESUMEN
BACKGROUND: Pure mucinous adenocarcinoma of the breast is a rare entity characterized by the production of variable amounts of mucin comprising 1% to 6% of breast carcinomas. Some mucinous adenocarcinomas have shown expression of intestinal differentiation markers such as MUC-2. This study examines the expression of intestinal differentiation markers in this type of breast carcinoma. RESULTS: Twenty-two cases of pure mucinous adenocarcinoma of the breast were assessed. Immunochemistry was performed for beta-catenin, CDX-2 and MUC-2. All cases were positive for B-catenin. MUC-2 positivity was observed in all cases; 63. 6% were 3 plus positive. All cases were negative for CDX-2. CONCLUSIONS: These results suggest that mucinous breast carcinomas express some markers of intestinal differentiation, such as MUC-2 and beta-catenin; however, future studies with a larger series of cases and using molecular techniques that help affirm these results are needed.
Asunto(s)
Adenocarcinoma Mucinoso/química , Biomarcadores de Tumor/análisis , Neoplasias de la Mama/química , Proteínas de Homeodominio/análisis , Mucosa Intestinal/química , Mucina 2/análisis , Transactivadores/análisis , beta Catenina/análisis , Adenocarcinoma Mucinoso/patología , Adulto , Anciano , Anciano de 80 o más Años , Antígenos de Diferenciación/análisis , Neoplasias de la Mama/patología , Factor de Transcripción CDX2 , Femenino , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
BACKGROUND: Pure mucinous adenocarcinoma of the breast is a rare entity characterized by the production of variable amounts of mucin comprising 1% to 6% of breast carcinomas. Some mucinous adenocarcinomas have shown expression of intestinal differentiation markers such as MUC-2. This study examines the expression of intestinal differentiation markers in this type of breast carcinoma. RESULTS: Twenty-two cases of pure mucinous adenocarcinoma of the breast were assessed. Immunochemistry was performed for beta-catenin, CDX-2 and MUC-2. All cases were positive for B-catenin. MUC-2 positivity was observed in all cases; 63. 6% were 3 plus positive. All cases were negative for CDX-2. CONCLUSIONS: These results suggest that mucinous breast carcinomas express some markers of intestinal differentiation, such as MUC-2 and beta-catenin; however, future studies with a larger series of cases and using molecular techniques that help affirm these results are needed.
Asunto(s)
Humanos , Femenino , Adulto , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Neoplasias de la Mama/química , Biomarcadores de Tumor/análisis , Transactivadores , Adenocarcinoma Mucinoso/química , Proteínas de Homeodominio/análisis , beta Catenina/análisis , Mucina 2/análisis , Mucosa Intestinal/química , Neoplasias de la Mama/patología , Inmunohistoquímica , Antígenos de Diferenciación/análisis , Estudios Retrospectivos , Adenocarcinoma Mucinoso/patología , Factor de Transcripción CDX2RESUMEN
Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.
Asunto(s)
Diferenciación Celular/efectos de los fármacos , Embrión de Mamíferos/efectos de los fármacos , Hepatocitos/citología , Hígado/citología , Células Madre/efectos de los fármacos , Animales , Antígenos de Diferenciación/análisis , Apolipoproteína B-100 , Apolipoproteínas B/aislamiento & purificación , Proliferación Celular , Dexametasona/administración & dosificación , Factores de Crecimiento de Fibroblastos/administración & dosificación , Violeta de Genciana , Glucógeno/metabolismo , Factor de Crecimiento de Hepatocito/administración & dosificación , Verde de Indocianina/farmacocinética , Ratones , Cultivo Primario de Células/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Madre/citología , Azul de Tripano , Tirosina Transaminasa/aislamiento & purificaciónRESUMEN
Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.
Asunto(s)
Animales , Ratones , Diferenciación Celular/efectos de los fármacos , Embrión de Mamíferos/efectos de los fármacos , Hepatocitos/citología , Hígado/citología , Células Madre/efectos de los fármacos , Antígenos de Diferenciación/análisis , Apolipoproteínas B/aislamiento & purificación , Proliferación Celular , Dexametasona/administración & dosificación , Factores de Crecimiento de Fibroblastos/administración & dosificación , Violeta de Genciana , Glucógeno/metabolismo , Factor de Crecimiento de Hepatocito/administración & dosificación , Verde de Indocianina/farmacocinética , Cultivo Primario de Células/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Madre/citología , Azul de Tripano , Tirosina Transaminasa/aislamiento & purificaciónRESUMEN
There are two hypotheses that explain tumor progression. The first one, the stochastic hypothesis, assumes that any cell within a tumor has the capacity to form and maintain the tumor mass. The second, the so-called hierarchical hypothesis, suggests the existence of a group of cells with a stem phenotype which, like in normal tissues, preserves tumors through a continuous production of progeny. These stem cells are in a particular niche, have a higher resistance to chemotherapy and radiotherapy, and are also capable of invading and migrating to other tissues. This review describes the cancer stem cells (CSCs), their function inside a tumor and the current knowledge about these cells.
Asunto(s)
Células Madre Neoplásicas , Antígenos de Diferenciación/análisis , Antígenos de Neoplasias/análisis , Biomarcadores de Tumor , División Celular , Linaje de la Célula , Separación Celular , Diseño de Fármacos , Resistencia a Antineoplásicos , Humanos , Modelos Biológicos , Invasividad Neoplásica , Metástasis de la Neoplasia , Células Neoplásicas Circulantes , Células Madre Neoplásicas/citología , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/efectos de la radiación , Tolerancia a RadiaciónRESUMEN
La leucemia de células dendríticas son patologías poco frecuentes, que involucran fundamentalmente a la piel, pero con una alta tendencia de metástasis. La inmunohistoquímica es una herramienta valiosa junto con la biopsia para el diagnóstico definitivo. Se presenta el caso de una paciente joven, quien acudió por presentar una lesión en la pierna izquierda de dimensiones y características únicas, cuyo diagnóstico requirió de la utilización de marcadores monoclonales específicos en la identificación de esta entidad.
The dendritic cell leukemias are pathologies very uncommon which involve mainly the skin, but with a high tendency of metastasis. The flow cytometric is a valuable tool together with tissue biopsy for a definitive diagnosis. We present the case of a young female who complained for a left leg lesion with unique dimensions and characteristics, in where the diagnostic process required the use of the specific monoclonal markers in the identification of this particular disease.
Asunto(s)
Humanos , Femenino , Adulto , Antígenos de Diferenciación/análisis , Células Dendríticas/fisiología , Células Dendríticas/patología , Leucemia/patología , Leucemia/tratamiento farmacológico , Traumatismos de la Pierna , Linfoma Cutáneo de Células T/patología , Tabaquismo/efectos adversos , Úlcera Cutánea/patología , Úlcera Cutánea/terapiaRESUMEN
INTRODUCTION: Oral lichen planus (OLP) is a relatively common inflammatory disease with a wide range of clinical forms. Its pathogenesis has not been fully elucidated although it is known to be mediated by lymphocytes with the participation of cytokines and other inflammatory cells, including type I and type II dermal dendrocytes (DD) (factor XIIIa+ DD and CD34+ DD, respectively). OBJECTIVES: To describe the presence and tissue distribution of these cells, through immunohistochemistry, in 23 specimens from patients with clinical and histopathological criteria of OLP. RESULTS: Factor XIIIa+ DD were mainly located in the superficial dermis (p < 0.0001) as opposed to the deep submucosa. These cells were abundant throughout the dermal-epidermal junction and closely related to lymphocyte infiltration. Moreover, factor XIIIa+ DD were also found in the epithelium and deep dermis. CD34+ DD were distributed mostly to the deep dermis directly below the lymphocyte infiltrate with few cells in the subepithelial region. CONCLUSIONS: DD were present in OLP, with distinct tissue distributions. Factor XIIIa+ DD were predominant in the superficial dermis while CD34+ DD could be found mostly in the deep dermis. These findings suggest that DD, and those positive for factor XIIIa+ in particular in view of their ability to express intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor alpha (TNF-alpha), may play an important role in pathogenesis of OLP.
Asunto(s)
Liquen Plano Oral/patología , Sistema Mononuclear Fagocítico/patología , Antígenos CD34/análisis , Antígenos de Diferenciación/análisis , Biopsia , Células de la Médula Ósea/citología , Linaje de la Célula , Factor XIIIa/análisis , Antígenos HLA-DR/análisis , Humanos , Liquen Plano Oral/inmunología , Sistema Mononuclear Fagocítico/química , Sistema Mononuclear Fagocítico/inmunología , Estudios RetrospectivosRESUMEN
Through the identification and subsequent targeting of an exquisitely unique and phenotypically defined cancer stem-cell population exhibiting discrete therapeutic vulnerabilities (a potential source of tumor recurrence) better survival rates for these patients may be achieved. It is this impetus that is making the field of pulmonary stem cell biology a growing field in biomedicine. These efforts are leading to the steady identification of multi-potent, self-renewing and proliferative progenitor cell populations throughout the bronchopulmonary tree. These cells give rise to both transiently amplifying (TA) and terminally differentiated (TD) cells, which (like in many other organs) are crucial for tissue homeostasis. In leukemia, it has been shown that partially committed cells, which are normally responsible for tissue maintenance after trauma, may undergo transformation via mutations resulting in the selective expression of genes that accentuate and perpetuate these cells' self-renewal capabilities. It is therefore perhaps legitimate to consider stem cells as protumorigenic. It is when these cells undergo genetic mutations which make them acquire the ability to metastasize, that cancer occurs, rendering the concept of "cancer stem cells" a rather attractive one indeed.
Asunto(s)
Transformación Celular Neoplásica/patología , Neoplasias Pulmonares/patología , Células Madre Neoplásicas/patología , Animales , Antígenos de Diferenciación/análisis , Biomarcadores de Tumor/análisis , Transformación Celular Neoplásica/genética , Predicción , Perfilación de la Expresión Génica , Proteínas Hedgehog/genética , Proteínas Hedgehog/fisiología , Humanos , Neoplasias Pulmonares/etiología , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia , Ratones , Ratones Endogámicos NOD , Ratones SCID , Mutación , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/fisiología , Trasplante de Neoplasias , Receptores Notch/genética , Receptores Notch/fisiología , Transducción de Señal , Células Madre/patología , Proteínas Wnt/genética , Proteínas Wnt/fisiologíaRESUMEN
INTRODUÇÃO: A galectina-3 (Gal-3) é uma lectina de mamíferos ligante de resíduos b-galactosídeos. Numerosos estudos têm mostrado que a Gal-3 apresenta importantes papéis na biologia tumoral, atuando em fenômenos como apoptose, metástase e transformação maligna. No entanto, em carcinomas de cabeça e pescoço, a real significância da sua expressão ainda é pouco compreendida. OBJETIVO: O objetivo deste estudo foi avaliar a expressão de Gal-3 em tumores de língua induzidos experimentalmente em camundongos desafiados com o carcinógeno 4-nitroquinolona-1-óxido (4NQO). MATERIAL E MÉTODOS: Quarenta e dois camundongos C57BL/6, machos, foram desafiados com 4NQO na água de beber por 16 semanas e sacrificados em diferentes períodos depois do tratamento. Após o sacrifício, as línguas foram removidas, processadas, coradas por hematoxilina e eosina (HE) e microscopicamente analisadas quanto à presença de carcinoma. Ensaio imuno-histoquímico para detecção do antígeno Gal-3 e análise descritiva da sua expressão nos tumores induzidos foram realizados. RESULTADOS: Ao final do experimento, foram produzidos 15 tumores. No tecido epitelial não-neoplásico, forte imunorreatividade foi observada apenas na camada parabasal. Nas camadas mais superficiais a intensidade de marcação foi mais fraca, e na camada basal variou de ausente a fraca. Todos os carcinomas bem diferenciados exibiram fraca marcação, exceto nas áreas queratinizantes. No único caso de carcinoma pouco diferenciado, forte imunorreatividade para Gal-3 foi observada. CONCLUSÃO: Nossos resultados descritivos mostram que a transformação maligna é acompanhada de redução da intensidade de expressão da Gal-3 e que o aumento da sua expressão com a perda da diferenciação neoplásica sugere a sua vinculação com agressividade tumoral.
BACKGROUND: Galectin-3 (Gal-3) is a b-galactoside-binding mammalian lectin. Numerous studies have demonstrated that Gal-3 plays an important role in tumor biology, acting in some events such as apoptosis, metastasis and malignant transformation. However, in carcinomas of head and neck, the real significance of Gal-3 expression requires a better understanding. OBJECTIVES: The aim of this paper was to evaluate Gal-3 expression in tongue carcinomas experimentally induced in mice challenged with carcinogen 4-nitroquinoline-1-oxide (4NQO). MATERIAL AND METHOD: Forty-two C57BL/6 male mice were challenged with 4NQO in drinking water for 16 weeks and killed at different periods after induction. In each period, their tongues were removed, routinely processed, stained with hematoxylin and eosin (H&E) and microscopically analyzed as to the presence of carcinoma. Immunohistochemical test for Gal-3 and a descriptive analysis of its expression in induced tumors were performed. RESULTS: By the end of the experiment, 15 tumors had been induced. In the non-neoplastic lingual epithelium, strong and weak immunoreactivity for Gal-3 was noted in parabasal and superficial layers, respectively. In the basal layer, Gal-3 expression varied from absent to weak. All the well-differentiated carcinomas showed weak immunoreactivity for Gal-3, except in keratinized areas. The only case of poorly differentiated squamous cell carcinoma indicated strong immunoreactivity for Gal-3. CONCLUSION: Our results show that malignant transformation is associated with reduced expression of Gal-3, whereas its increased expression in poorly differentiated carcinoma seems to be connected with tumor aggressiveness.
Asunto(s)
Animales , Masculino , Ratones , Carcinoma de Células Escamosas/inducido químicamente , /análisis , /inmunología , Neoplasias de la Lengua/inducido químicamente , Antígenos de Diferenciación/análisis , Inmunohistoquímica , Modelos Animales , Biomarcadores de Tumor , Transformación Celular Neoplásica/inmunologíaRESUMEN
Pleomorphic adenoma (PA) and adenoid cystic carcinoma (ACC) are the commonest benign and malignant salivary gland tumours respectively. Interactions between cells and extracellular matrix of PA and ACC, partially mediated by integrins, are important in their biology. The expression of integrins is regulated by numerous factors, amongst them, transforming growth factor beta1 (TGFbeta1). Our study investigated the effects of TGFbeta1 on the expression of integrin beta subunits in vitro and on the expression of cytoskeletal proteins of cells derived from PA and ACC. The expression of cytoskeletal differentiation markers and integrins was assessed using immunofluorescence. ELISA assays were employed to quantitate the expression integrins and MTT assays evaluated the mitochondrial activity of cells stimulated with TGFbeta1. PA cells showed increased expression of integrins and de novo expression of differentiation markers upon TGFbeta1 stimulation. ACC cells were less responsive to such stimulation. This may reflect important differences in the biological behaviour of benign and malignant cells.
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Adenoma Pleomórfico/metabolismo , Carcinoma Adenoide Quístico/metabolismo , Integrinas/metabolismo , Neoplasias de la Parótida/metabolismo , Factor de Crecimiento Transformador beta1/farmacología , Adulto , Antígenos de Diferenciación/análisis , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Citoesqueleto/metabolismo , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Integrina beta1/análisis , Integrina beta1/metabolismo , Integrina beta3/análisis , Integrina beta3/metabolismo , Integrina beta4/análisis , Integrina beta4/metabolismo , Integrinas/análisis , Persona de Mediana Edad , Estimulación Química , Células Tumorales CultivadasRESUMEN
The long-term persistence of pathogens in a host is a hallmark of certain infectious diseases, including schistosomiasis, leishmaniasis, and paracoccidioidomycosis (PCM). Natural regulatory T (Treg) cells are involved in control of the immune responses, including response to pathogens. Because CTLA-4 is constitutively expressed in Treg cells and it acts as a negative regulator of T cell activation in patients with PCM, here we investigated the involvement of Treg cells in the control of systemic and local immune response in patients with PCM. We found that the leukocyte subsets were similar in patients and controls, except for CD11c+CD1a+ cells. However, a higher frequency of CD4+CD25+ T cells expressing CTLA-4, glucorticoid-inducible TNFR, membrane-bound TGF-beta, and forkhead-box 3 were observed in PBMC of patients. In accordance, these cells exhibited stronger suppressive activity when compared with those from controls (94.0 vs 67.5% of inhibition of allogeneic T cell proliferation). In addition, the data showed that CD4+CD25+ T cells expressing CTLA-4+, glucocorticoid-inducible TNFR positive, CD103+, CD45RO+, membrane-bound TGF-beta, forkhead-box 3 positive, and the chemokines receptors CCR4 and CCR5 accumulate in the Paracoccidioides brasiliensis-induced lesions. Indeed, the secreted CCL17 and CCL22, both associated with the migration of Treg cells to peripheral tissues, were also detected in the biopsies. Moreover, the CD4+CD25+ T cell derived from lesions, most of them TGF-beta+, also exhibited functional activity in vitro. Altogether, these data provide the first evidence that Treg cells play a role in controlling local and systemic immune response in patients with a fungal-induced granulomatous disease advancing our understanding about the immune regulation in human chronic diseases.
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Antígenos CD4/análisis , Subunidad alfa del Receptor de Interleucina-2/análisis , Paracoccidioidomicosis/inmunología , Linfocitos T Reguladores/inmunología , Antígenos CD/análisis , Antígenos de Diferenciación/análisis , Antígeno CTLA-4 , Membrana Celular/química , Membrana Celular/inmunología , Movimiento Celular , Quimiocina CCL17 , Quimiocina CCL22 , Quimiocinas/metabolismo , Quimiocinas CC/metabolismo , Enfermedad Crónica , Citocinas/metabolismo , Proteína Relacionada con TNFR Inducida por Glucocorticoide , Factor Nuclear 3-gamma del Hepatocito/análisis , Humanos , Cadenas alfa de Integrinas/análisis , Antígenos Comunes de Leucocito/análisis , Paracoccidioidomicosis/patología , Fenotipo , Receptores CCR4 , Receptores CCR5/análisis , Receptores de Quimiocina/análisis , Receptores de Factor de Crecimiento Nervioso/análisis , Receptores del Factor de Necrosis Tumoral/análisis , Factor de Crecimiento Transformador beta/análisisRESUMEN
During helminthic infections, strong Th2 type-biased responses concomitant with impaired cell-proliferative responses to parasitic and unrelated antigens are major immunological hallmarks. Parasite glycan structures have been proposed to play a role in modulating these responses. To understand early events related to immune modulation during cestode infection, we have examined the role of intact glycans of antigens from Taenia crassiceps in the recruitment of innate cells. Soluble antigens from this cestode contained higher levels of carbohydrates than proteins. Intraperitoneal injection of the antigens rapidly recruited a cell population expressing F4/80(+)/Gr-1(+)surface markers, which adoptively suppressed naïve T-cell proliferation in vitro in response to anti-CD3/CD28 MAb stimulation in a cell-contact dependent manner. Soluble antigens with altered glycans by treatment with sodium periodate significantly reduced the recruitment of F4/80(+)/Gr1(+)cells, concomitantly their suppressive activity was abrogated, indicating that glycans have a role in the early activation of these suppressor cells. Using C3H/HeJ and STAT6-KO mice, we found that expansion and suppressive activity of F4/80(+)Gr1(+)cells induced by T. crassiceps intact antigens was TLR4 and Th2-type cytokine independent. Together with previous studies on nematode and trematode parasites, our data support the hypothesis that glycans can be involved on a similar pathway in the immunoregulation by helminths.
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Antígenos Helmínticos/inmunología , Cestodos/inmunología , Infecciones por Cestodos/inmunología , Células Mieloides/inmunología , Polisacáridos/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Antígenos de Diferenciación/análisis , Antígenos Helmínticos/química , Antígenos Helmínticos/aislamiento & purificación , Antígenos CD28/inmunología , Complejo CD3/inmunología , Técnicas de Cocultivo , Citocinas/inmunología , Femenino , Citometría de Flujo , Ratones , Receptores de Quimiocina/análisis , Receptor Toll-Like 4/inmunologíaRESUMEN
PROBLEM: It has been suggested that type of chimeric mRNA is associated with differences in the clinical and hematologic characteristics of chronic myeloid leukemia (CML). However, prognostic value of type of chimeric mRNA bcr-xabl (b3a2 or b2a2) is still controversial. METHODS: We analyzed 97 cases of Philadelphia-positive CML to determine mRNA type by reverse-polymerase chain reaction (RT-PCR) and its relationship with clinical features. RESULTS: We detected b3a2 bcr-abl transcripts in 27 (28%) cases, b2a2 in 57 (59%) cases, and 13 (13%) with both mRNA transcripts b3a2/b2a2. These frequencies were the total reverse of other reports. Age, sex, hemoglobin, and white-cell counts showed no significant difference for those with either b3a2 or b2a2 bcr-abl transcripts. However, platelet counts of b3a2 patients were significantly higher than those of b2a2 patients (743.3 vs 477.3 x 109/L; p = 0.01). In addition, in the subgroup of patients whose white-cell count at diagnosis was < 100 x 10(9)/L, those with b3a2 transcript had a significantly higher platelet count (679.1 vs. 352.2 x 10(9)/L; p = 0.001). CONCLUSIONS: We observed reversed frequency of bcr-abl transcripts in this population, but agreement with other Latin-American reports. In addition, our data suggested that there is different CML biological behavior in our population and that there is a subpopulation of CML patients in whom b3a2 is associated witH higher thrombopoietic activity.
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Antígenos CD , Proteínas de Fusión bcr-abl/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Glicoproteínas de Membrana , Recuento de Plaquetas , Factores de Transcripción/genética , Adolescente , Adulto , Anciano , Antígenos de Diferenciación/análisis , Antígenos de Diferenciación/genética , Proteína BRCA2/análisis , Proteína BRCA2/genética , Antígeno CD24 , Femenino , Proteínas de Fusión bcr-abl/análisis , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/sangre , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Transcripción/químicaRESUMEN
Galectin-3 is a protein of the lectin family that has been associated with neoplastic processes in various tissues. In the thyroid, expression of this protein has been described in differentiated follicular cancer, suggesting that the immunohistochemical study of galectin-3 may be a potential marker of malignancy in thyroid neoplasms. The confirmation of these results may represent an extremely useful tool for presurgical diagnosis and medical conduct. In this study, galectin-3 protein and mRNA expression were analyzed in the thyroid tissues from 87 patients with histomorphological diagnosis of multinodular goiter (MNG) (n = 24), follicular adenoma (n = 31), follicular carcinoma (n = 20), papillary carcinoma (n = 12), and five normal tissues. Galectin-3 protein expression was detected by immunohistochemical method in light, fluorescence, and confocal microscopy, using monoclonal antibody. Galectin-3 mRNA expression was detected by the RT-PCR method. Our results showed that the majority of carcinomas expressed galectin-3 protein (follicular, 90%; papillary, 100%). However, in contrast to the previously published data, benign lesions also expressed galectin-3 (adenoma, 45%; MNG, 17%). We further demonstrated by RT-PCR that thyroid tissues with diagnosis of adenoma and MNG-expressed galectin-3 mRNA. Although the galectin-3 immunostaining demonstrated a sensitivity of 93.8% in the identification of cancer, the accuracy in the distinction between benign and malignant tissues was 77.0%. This accuracy was even lower (68.6%) when the galectin-3 expression in follicular adenoma was compared with follicular carcinoma. Thus, the use of galectin-3 immunodetection as a molecular marker for thyroid carcinoma must be interpreted with caution, particularly in the differentiation between thyroid follicular carcinoma and follicular adenoma.
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Antígenos de Diferenciación/análisis , Antígenos de Diferenciación/genética , ARN Mensajero/análisis , Neoplasias de la Tiroides/química , Adenocarcinoma Folicular/química , Adenoma/química , Biomarcadores de Tumor/análisis , Carcinoma Papilar/química , Galectina 3 , Bocio/metabolismo , Humanos , Inmunohistoquímica , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sensibilidad y Especificidad , Glándula Tiroides/químicaRESUMEN
The cellular immune response probably plays a pivotal role in determining the clinical outcome after exposure to Mycobacterium tuberculosis. We used multi-parameter flow-cytometry to evaluate the distribution of T-lymphocyte subsets during infection and disease caused by M. tuberculosis. Samples were obtained from 71 volunteers to identify the T CD4+ and CD8+ lymphocyte numbers, and the activation plus memory/naïve phenotypes, as defined by CD38, HLA-DR, CD45RA and CD27 markers. Subjects were divided into 18 healthy volunteers without detectable reaction to purified protein derivative (PPD-), 18 health care workers with a recent conversion to PPD, 20 patients with active pulmonary tuberculosis (TBC) and 15 patients with treated TBC at 6 months of therapy. By multiple-comparison analyses, the T CD4+ lymphocyte number of the TBC group was lower than the PPD- group (P < 0.05). This difference was apparently lost after treatment. The higher and the lower number of naïve T CD4+ cells was observed in the PPD- and TBC group, respectively. CD8+ T lymphocytes were also statistically different among the four groups (P = 0.0002), lower in the TBC group (P < 0.05). CD8+ T lymphocyte activation was evaluated by the CD38 and HLA-DR surface expression. The percentage distribution of these markers was statistically different between the four groups (P = 0.0055). TBC patients had a higher percentage of CD38+ cells and mean fluorescence index, suggesting an overall increase of cell activation. These results suggest that peripheral T lymphocytes reflect cellular activation during TBC, along with possible redistribution of naïve, memory/effector and late differentiated memory/effector phenotypes in the peripheral blood after infection and disease caused by M. tuberculosis.