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1.
Braz. J. Pharm. Sci. (Online) ; 58: e19685, 2022. graf
Artículo en Inglés | LILACS | ID: biblio-1383966

RESUMEN

Abstract Psoriasis is a chronic skin inflammation, characterized by impaired differentiation, hyperproliferation of keratinocytes involving pro-inflammatory factors interleukin (IL)-13/17A, tumor necrosis factor (TNF)-α, interferon (IFN)-γ. Among the integrin family, α5 is important for blood vessel formation, and ß4 for proliferation, differentiation of keratinocytes. To investigate the expression and regulation of integrin α5 and ß4 in psoriatic keratinocytes. Skin biopsies were obtained from 14 psoriatic patients and 12 normal volunteers. We compared the immunolocalization and regulation of α5 and ß4 between the psoriatic and normal ones, before and after incubation with MEK/ERK pathway inhibitor U0126 by immunohistochemistry and western blot separately. Immunohistochemistry showed psoriatic keratinocytes had higher α5 than normal ones. According to western blot, IL-17A and IL-13 increased normal keratinocytes' α5 and ß4 respectively, but psoriatic keratinocytes were the exact opposite. Incubated with U0126, normal keratinocytes' α5 was enhanced by the 5 cytokines ; while IL-13/17A, IFN-γ suppressed ß4. Psoriatic keratinocytes' α5 was increased by IL-13/17A, decreased by IFN-γ; but ß4 increased by IL-17A, IFN-γ. IL-13/17A, TNF-α, IFN-γ regulate α5 and ß4 through ERK pathway whether normal or psoriasis. The normal and psoriatic keratinocytes respond to the same cytokines differently


Asunto(s)
Humanos , Masculino , Femenino , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Integrinas/análisis , Queratinocitos/clasificación , Pacientes/clasificación , Psoriasis/patología , Western Blotting/instrumentación , Citocinas/agonistas , Interleucinas/análisis
2.
J Appl Oral Sci ; 28: e20190156, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32049134

RESUMEN

OBJECTIVE: The present study aimed to investigate the participation of focal adhesion kinases (FAK) in interactions between osteoblastic cells and titanium (Ti) surfaces with three different topographies, namely, untreated (US), microstructured (MS), and nanostructured (NS). METHODOLOGY: Osteoblasts harvested from the calvarial bones of 3-day-old rats were cultured on US, MS and NS discs in the presence of PF-573228 (FAK inhibitor) to evaluate osteoblastic differentiation. After 24 h, we evaluated osteoblast morphology and vinculin expression, and on day 10, the following parameters: gene expression of osteoblastic markers and integrin signaling components, FAK protein expression and alkaline phosphatase (ALP) activity. A smooth surface, porosities at the microscale level, and nanocavities were observed in US, MS, and NS, respectively. RESULTS: FAK inhibition decreased the number of filopodia in cells grown on US and MS compared with that in NS. FAK inhibition decreased the gene expression of Alp, bone sialoprotein, osteocalcin, and ALP activity in cells grown on all evaluated surfaces. FAK inhibition did not affect the gene expression of Fak, integrin alpha 1 ( Itga1 ) and integrin beta 1 ( Itgb1 ) in cells grown on MS, increased the gene expression of Fak in cells grown on NS, and increased the gene expression of Itga1 and Itgb1 in cells grown on US and NS. Moreover, FAK protein expression decreased in cells cultured on US but increased in cells cultured on MS and NS after FAK inhibition; no difference in the expression of vinculin was observed among cells grown on all surfaces. CONCLUSIONS: Our data demonstrate the relevance of FAK in the interactions between osteoblastic cells and Ti surfaces regardless of surface topography. Nanotopography positively regulated FAK expression and integrin signaling pathway components during osteoblast differentiation. In this context, the development of Ti surfaces with the ability to upregulate FAK activity could positively impact the process of implant osseointegration.


Asunto(s)
Proteína-Tirosina Quinasas de Adhesión Focal/antagonistas & inhibidores , Osteoblastos/efectos de los fármacos , Quinolonas/farmacología , Sulfonas/farmacología , Titanio/química , Animales , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Proteína-Tirosina Quinasas de Adhesión Focal/análisis , Proteína-Tirosina Quinasas de Adhesión Focal/química , Expresión Génica , Integrinas/análisis , Microscopía Electrónica de Rastreo , Oseointegración/efectos de los fármacos , Osteoblastos/fisiología , Quinolonas/química , Ratas Wistar , Reacción en Cadena en Tiempo Real de la Polimerasa , Transducción de Señal , Sulfonas/química , Propiedades de Superficie
3.
J. appl. oral sci ; J. appl. oral sci;28: e20190156, 2020. graf
Artículo en Inglés | LILACS, BBO - Odontología | ID: biblio-1090765

RESUMEN

Abstract Objective The present study aimed to investigate the participation of focal adhesion kinases (FAK) in interactions between osteoblastic cells and titanium (Ti) surfaces with three different topographies, namely, untreated (US), microstructured (MS), and nanostructured (NS). Methodology Osteoblasts harvested from the calvarial bones of 3-day-old rats were cultured on US, MS and NS discs in the presence of PF-573228 (FAK inhibitor) to evaluate osteoblastic differentiation. After 24 h, we evaluated osteoblast morphology and vinculin expression, and on day 10, the following parameters: gene expression of osteoblastic markers and integrin signaling components, FAK protein expression and alkaline phosphatase (ALP) activity. A smooth surface, porosities at the microscale level, and nanocavities were observed in US, MS, and NS, respectively. Results FAK inhibition decreased the number of filopodia in cells grown on US and MS compared with that in NS. FAK inhibition decreased the gene expression of Alp, bone sialoprotein, osteocalcin, and ALP activity in cells grown on all evaluated surfaces. FAK inhibition did not affect the gene expression of Fak, integrin alpha 1 ( Itga1 ) and integrin beta 1 ( Itgb1 ) in cells grown on MS, increased the gene expression of Fak in cells grown on NS, and increased the gene expression of Itga1 and Itgb1 in cells grown on US and NS. Moreover, FAK protein expression decreased in cells cultured on US but increased in cells cultured on MS and NS after FAK inhibition; no difference in the expression of vinculin was observed among cells grown on all surfaces. Conclusions Our data demonstrate the relevance of FAK in the interactions between osteoblastic cells and Ti surfaces regardless of surface topography. Nanotopography positively regulated FAK expression and integrin signaling pathway components during osteoblast differentiation. In this context, the development of Ti surfaces with the ability to upregulate FAK activity could positively impact the process of implant osseointegration.


Asunto(s)
Animales , Osteoblastos/efectos de los fármacos , Sulfonas/farmacología , Titanio/química , Quinolonas/farmacología , Proteína-Tirosina Quinasas de Adhesión Focal/antagonistas & inhibidores , Osteoblastos/fisiología , Sulfonas/química , Propiedades de Superficie , Microscopía Electrónica de Rastreo , Transducción de Señal , Expresión Génica , Integrinas/análisis , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Oseointegración/efectos de los fármacos , Ratas Wistar , Quinolonas/química , Proliferación Celular/efectos de los fármacos , Proteína-Tirosina Quinasas de Adhesión Focal/análisis , Proteína-Tirosina Quinasas de Adhesión Focal/química , Reacción en Cadena en Tiempo Real de la Polimerasa
4.
Curr Top Med Chem ; 19(22): 2069-2078, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31385773

RESUMEN

BACKGROUND: Disintegrins from snake venoms bind with high specificity cell surface integrins, which are important pharmacological targets associated with cancer development and progression. OBJECTIVE: In this study, we isolated a disintegrin from the Porthidium lansbergii lansbergii venom and evaluated its antitumoral effects on breast cancer cells. METHODS: The isolation of the disintegrin was performed on RP-HPLC and the inhibition of platelet aggregation was evaluated on human platelet-rich plasma. The inhibition of cell adhesion was also evaluated in vitro on cultures of cell lines by the MTT method as well as the inhibition of breast cancer cell migration by the wound healing assay. The binding of the disintegrin to integrin subunits was verified by flow cytometry and confocal microscopy. Finally, inhibition of angiogenesis was assessed in vitro on HUVEC cells and the concentration of VEGF was measured in the cellular supernatants. RESULTS: The disintegrin, named Lansbermin-I, is a low molecular weight protein (< 10 kDa) that includes an RGD on its sequence identified previously. Lansbermin-I showed potent inhibition of ADP and collagen-induced platelet aggregation on human plasma and also displayed inhibitory effects on the adhesion and migration of breast cancer MCF7 and MDA-MB 231cell lines, without affecting nontumorigenic breast MCF-10A and lung BEAS cells. Additionally, Lansbermin-I prevented MCF7 cells to adhere to fibronectin and collagen, and also inhibited in vitro angiogenesis on human endothelial HUVEC cells. CONCLUSION: Our results display the first report on the antitumor and anti-metastatic effects of an RGDdisintegrin isolated from a Porthidium snake venom by possibly interfering with α2 and/or ß1-containing integrins. Thus, Lansbermin-I could be an attractive model to elucidate the role of disintegrins against breast cancer development.


Asunto(s)
Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Venenos de Crotálidos/farmacología , Desintegrinas/farmacología , Animales , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Adhesión Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Venenos de Crotálidos/química , Venenos de Crotálidos/aislamiento & purificación , Desintegrinas/química , Desintegrinas/aislamiento & purificación , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Integrinas/análisis , Integrinas/metabolismo , Neovascularización Patológica/tratamiento farmacológico , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Relación Estructura-Actividad , Viperidae , Cicatrización de Heridas/efectos de los fármacos
5.
Artif Organs ; 43(8): 764-772, 2019 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-30779451

RESUMEN

A titanium surface nitrided by plasma contains nitrogen ions that guarantee resistance to corrosion and biocompatibility. Despite this, no descriptions concerning the influence of the expression of cell adhesion proteins and their influence on osteogenic cell differentiation are available. Thus, the present study aimed to assess the response of murine pre-osteoblastic cells (MC3T3-E1) cultured on nitrided titanium surfaces. Pre-osteoblastic cells were grown on polished titanium discs, used as controls, and on previously characterized plasma-nitrided titanium discs. Cells from both groups were submitted to the MTT cell viability test. The expressions of α5, α2, and ß1 integrin were assessed by flow cytometry and immunofluorescence, while osteocalcin expression was assessed by flow cytometry. The nitrided surface presented higher α2 and ß1 integrin expressions, as well as osteocalcin expression, when compared to the polished surface, with no alterations in cell viability. These findings seem to suggest that the plasma nitriding treatment produces a titanium surface with the potential for effective in vitro osseointegration.


Asunto(s)
Materiales Biocompatibles/química , Osteoblastos/citología , Gases em Plasma/química , Titanio/química , Animales , Adhesión Celular , Diferenciación Celular , Línea Celular , Corrosión , Integrinas/análisis , Ratones , Oseointegración , Osteogénesis , Propiedades de Superficie
6.
Am J Dermatopathol ; 35(5): 541-54, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23715077

RESUMEN

Primary oral mucosal melanoma is a rare aggressive tumor. Recent studies have demonstrated a correlation between increased tumor invasion and the metastatic phenotype and altered adhesion molecule expression profiles. The present study analyzed the expression of integrins, claudins, and immunoglobulin-like adhesion molecules in oral mucosal melanomas and correlated results with clinical parameters. Immunohistochemical analyses of the expression patterns of these molecules were performed on thirty-five cases of primary oral mucosal melanomas organized in a tissue microarray. The results were correlated with clinical and histological features of the cohort. A number of integrin subunits were negative and this was related with vascular invasion. Positivity of integrin beta-3 and CD166 (activated leukocyte cell adhesion molecule) was statistically associated with extensive vascular invasion (P < 0.05). Lower expression of CD54 (intercellular cell adhesion molecule) was associated with cases with extensive necrosis. Most cases with metastatic disease were negative for CD66 (carcinoembryonic antigen-related cell adhesion molecule). Several subunits of claudins were negative and, although not statistically significant, this lack of expression was partially associated with histological factors of poor prognosis. Altered patterns of adhesion molecule expression, mainly integrins and immunoglobulin-like proteins, may participate in the pathogenesis and outcome of oral mucosal melanomas.


Asunto(s)
Biomarcadores de Tumor/análisis , Moléculas de Adhesión Celular/análisis , Claudinas/análisis , Inmunoglobulinas/análisis , Integrinas/análisis , Melanoma/química , Mucosa Bucal/química , Neoplasias de la Boca/química , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Bolivia , Brasil , Distribución de Chi-Cuadrado , Niño , Preescolar , Femenino , Humanos , Inmunohistoquímica , Lactante , Recién Nacido , Masculino , Melanoma/secundario , Persona de Mediana Edad , Mucosa Bucal/patología , Neoplasias de la Boca/patología , Necrosis , Clasificación del Tumor , Invasividad Neoplásica , Pronóstico , Análisis de Matrices Tisulares , Adulto Joven
7.
Pathol Res Pract ; 208(10): 598-603, 2012 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-22917688

RESUMEN

The progression of carcinogenesis entails the detachment of cells, invasion and migration of neoplastic cells. Alterations in epithelial adhesion and basement membrane proteins might mediate the early stages of carcinogenesis. This study investigated the expression of adhesion molecules and the basement membrane protein laminin-5 in actinic cheilitis (AC) and incipient squamous cell carcinoma of the lower lip to understand early photocarcinogenesis. Ln-5γ2 chain as well as α3, ß1 subunits of α3ß1 heterodimer and ß4 subunit of integrin α6ß4 were evaluated by immunohistochemistry in 16 cases of AC and 16 cases of superficially invasive squamous cell carcinoma (SISCC). Most AC cases showed reduced expression of ß1, ß4 and α3 integrins, and SISCCs lacked ß1, ß4 and α3 integrins in the invasive front. AC cases were negative for the Ln-5γ2 chain. Five cases of SISCC (31%) showed heterogeneous Ln-5γ2 chain expression in the invasive front of the tumor. Integrin ß1, ß4 and α3 expression is lost during the early stages of lip carcinogenesis. Expression of Ln-5γ2 in the invasive front in cases and its correlation with tumor progression suggest that it mediates the acquisition of the migrating and invading epithelial cell phenotype.


Asunto(s)
Biomarcadores de Tumor/análisis , Carcinoma de Células Escamosas/química , Queilitis/metabolismo , Integrinas/análisis , Laminina/análisis , Neoplasias de los Labios/química , Labio/química , Biopsia , Carcinoma de Células Escamosas/patología , Adhesión Celular , Movimiento Celular , Queilitis/patología , Humanos , Inmunohistoquímica , Integrina alfa3/análisis , Integrina beta1/análisis , Integrina beta4/análisis , Labio/patología , Neoplasias de los Labios/patología , Invasividad Neoplásica , Fenotipo
8.
Artículo en Inglés | MEDLINE | ID: mdl-17942347

RESUMEN

OBJECTIVE AND STUDY DESIGN: The morphological distinction between incipient dentigerous cyst (IDC) and enlarged pericoronal dental follicle (EDF) remains one of the most controversial questions in the literature. The objective of this study was to analyze the immunohistochemistry expression of alfa(2)beta(1), alfa(3)beta(1), and alfa(5)beta(1) in 23 cases of EDFs and 21 cases of IDCs. RESULTS: All integrins were immunopositive in the cases studied. A significant difference was detected regarding alfa(2)beta(1) integrin (P < .0001) in which a higher expression was present in IDCs. Moreover, statistical difference was also found between basal and suprabasal cell layer in cystic epithelium (P < .0034). The alfa(3)beta(1) integrin expression showed significant difference (P < .013) between EDF and IDC with a tendency of more pronounced staining in IDC. CONCLUSIONS: These results corroborate the possibility of histopathological distinction between EDF and IDC in which squamous metaplasia of reduced enamel epithelium to stratified epithelium would be the first event of cystic transformation.


Asunto(s)
Saco Dental/patología , Quiste Dentígero/patología , Integrinas/análisis , Saco Dental/química , Saco Dental/diagnóstico por imagen , Quiste Dentígero/química , Quiste Dentígero/diagnóstico por imagen , Humanos , Radiografía , Coloración y Etiquetado/métodos
9.
Int J Exp Pathol ; 88(3): 191-8, 2007 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-17504449

RESUMEN

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.


Asunto(s)
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 Cultivadas
10.
Biochem Biophys Res Commun ; 343(1): 326-33, 2006 Apr 28.
Artículo en Inglés | MEDLINE | ID: mdl-16540081

RESUMEN

Sperm-oocyte plasma membrane interaction is an essential step in fertilization. In amphibians, the molecules involved have not been identified. Our aim was to detect and characterize oocyte molecules with binding affinity for sperm. We isolated plasma membranes free from vitelline envelope and yolk proteins from surface-biotinylated Bufo arenarum oocytes. Using binding assays we detected a biotinylated 100 kDa plasma membrane protein that consistently bound to sperm. Chromatographic studies confirmed the 100 kDa protein and detected two additional oocyte molecules of 30 and 70 kDa with affinity for sperm. Competition studies with an integrin-interacting peptide and cross-reaction with an anti-HSP70 antibody suggested that the 100 and 70 kDa proteins are members of the integrin family and HSP70, respectively. MS/MS analysis suggested extra candidates for a role in this step of fertilization. In conclusion, we provide evidence for the involvement of several proteins, including integrins and HSP70, in B. arenarum sperm-oocyte plasma membrane interactions.


Asunto(s)
Bufo arenarum/metabolismo , Proteínas HSP70 de Choque Térmico/análisis , Integrinas/análisis , Proteínas de la Membrana/análisis , Oocitos/química , Interacciones Espermatozoide-Óvulo , Animales , Membrana Celular/química , Membrana Celular/metabolismo , Femenino , Proteínas HSP70 de Choque Térmico/metabolismo , Integrinas/metabolismo , Masculino , Proteínas de la Membrana/metabolismo , Oocitos/metabolismo , Espermatozoides/metabolismo
11.
São Paulo; s.n; 2006. 61 p. ilus. (BR).
Tesis en Portugués | LILACS, BBO - Odontología | ID: lil-436130

RESUMEN

Diversos estudos têm demonstrado que os implantes de titânio são altamente biocompatíveis e passíveis de osseointegrar, contudo os mecanismos moleculares que atuam por trás da osseointegração permanecem largamente inexplorados. Uma das possibilidades é que o implante exposto ao sangue do paciente durante a cirurgia absorve proteínas relacionadas com a adesão celular, como a fibronectina e vitronectina presentes no plasma. Células como osteoblastos podem então aderir a estas proteínas através dos mecanismos mediadas pelas integrinas. No presente estudo, utilizamos a imunofluorescência para marcação dos anticorpos contra integrinas α2, α3, α5, α6, αv e β1 em células OsA-CL cultivadas sobre lamínulas de vidro e superfície de titânio modificada por radiação à laser no período de 1h à 24 horas. As células aderidas na superfície lisa das lamínulas de vidro tiveram um maior espalhamento durante as primeiras 3 horas, porém nos outros períodos estudados o espalhamento ocorreu de maneira similar em ambas as superfícies. A expressão de integrinas na superfície rugosa dos implantes manteve-se uniforme em todos os períodos estudados, contudo no grupo controle a expressão de integrinas diminuiu na avaliação de 24 horas. Concluiu-se que as características de superfície dos diferentes biomateriais podem afetar o comportamento celular durante a interação inicial das células com a superfície de material utilizado como substrato


Asunto(s)
Materiales Biocompatibles , Implantes Dentales , Integrinas/análisis , Rayos Láser , Patología Bucal
12.
J Oral Pathol Med ; 33(9): 574-80, 2004 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-15357679

RESUMEN

BACKGROUND: Pleomorphic adenoma (PA) and adenoid cystic carcinoma (ACC) are neoplasms of distinct behaviour, showing similar origin, cell components and marked presence of extracellular matrix (ECM). Interactions between cells and ECM are important in the biology of tumours, being partially mediated by integrins. This study investigated these interactions on PA and ACC using paraffin-embedded tissue and an in vitro model of these conditions. METHODS: Expression of integrins in paraffin-embedded samples was assessed by immunohistochemistry. Cells from PA and ACC were characterized using immunofluorescence, and integrin patterns of expression were investigated on cells cultivated on different ECM proteins. RESULTS: Luminal cells of both PA and ACC were more intensely positive for integrins than myoepithelial cells. In vitro studies revealed that PA cells expressed more integrins than ACC cells regardless the ECM protein present. CONCLUSIONS: This study revealed particular patterns of integrin expression in both specimens and in vitro models of PA and ACC. This might prove useful for a better understanding of the biology of these lesions.


Asunto(s)
Adenoma Pleomórfico/patología , Carcinoma Adenoide Quístico/patología , Matriz Extracelular/patología , Integrinas/análisis , Actinas/análisis , Proteínas de Unión al Calcio/análisis , Proteínas de Unión a Calmodulina/análisis , Colágeno Tipo I/análisis , Colágeno Tipo IV/análisis , Fibroblastos/patología , Fibronectinas/análisis , Humanos , Integrina alfa2/análisis , Integrina alfa3/análisis , Integrina alfa5/análisis , Integrina beta1/análisis , Laminina/análisis , Proteínas de Microfilamentos , Células del Estroma/patología , Vimentina/análisis , Calponinas
13.
Braz J Med Biol Res ; 36(8): 1101-9, 2003 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-12886465

RESUMEN

Mast cell progenitors arise in bone marrow and then migrate to peripheral tissues where they mature. It is presumed that integrin receptors are involved in their migration and homing. In the present study, the expression of various integrin subunits was investigated in three systems of adherent and nonadherent mast cells. Mesentery mast cells, freshly isolated bone marrow-derived mast cells (BMMC) and RBL-2H3 cells grown attached to tissue culture flasks are all adherent mast cells and peritoneal mast cells, and cultured BMMC and RBL-2H3 cells grown in suspension represent nonadherent mast cell populations. Pure populations of mast cells were immunomagnetically isolated from bone marrow, mesentery and peritoneal lavage using the mast cell-specific monoclonal antibody AA4. By immunomicroscopy, we could demonstrate that all of these mast cells expressed alpha 4, alpha 5, alpha 6, beta 1 and beta 7 integrin subunits. The expression of the alpha 4 integrin subunit was 25% higher in freshly isolated mesentery mast cells and BMMC. Consistent with the results obtained by immunomicroscopy, mesentery mast cells expressed 65% more mRNA for the alpha 4 integrin subunit than peritoneal mast cells. In vitro studies were also conducted using the rat mast cell line RBL-2H3. RBL-2H3 cells grown attached to the tissue culture flasks or as suspension cultures expressed the same integrin subunits identified in bone marrow, mesenteric and peritoneal mast cells ex vivo. Similarly, the expression of alpha 4 integrin was higher in adherent cells. Therefore, alpha 4 integrins may play a critical role in the anchorage of mast cells to the extracellular matrix in bone marrow and in peripheral tissues.


Asunto(s)
Movimiento Celular , Integrinas/análisis , Mastocitos/química , Proteínas de Neoplasias/análisis , Animales , Adhesión Celular , Células Cultivadas , Regulación de la Expresión Génica , Masculino , Mastocitos/fisiología , Ratones , Ratones Endogámicos BALB C , Ratas , Ratas Wistar
14.
Virology ; 305(1): 93-105, 2003 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-12504544

RESUMEN

In vivo replication of rotaviruses is generally limited to enterocytes. Because of this restriction, most blood circulating rotavirus-specific B cells are hypothesized to originate in Peyer's patches and should express the intestinal homing receptor alpha4beta7. To test this hypothesis in humans, we used a flow cytometry assay that identifies antigen-activated (IgD-) B cells (CD19+) that express surface rotavirus-specific immunoglobulin. With this assay we could detect rotavirus-specific B cells in both children and adults with an acute rotavirus (RV) infection. Staining with an anti-alpha4beta7 monoclonal antibody, we could determine that B cells that express rotavirus-specific surface immunoglobulin predominantly express alpha4beta7. The response of rotavirus-specific antibody-secreting cells in the peripheral blood of children and adults with acute rotavirus infection was also studied by ELISPOT. The antibody-secreting cells of children were mainly of the IgM isotype, while the antibody-secreting cells of adults were predominantly of the IgA and IgG isotype. alpha4beta7+ and alpha4beta7- subsets of peripheral blood mononuclear cells were purified using paramagnetic beads and then tested in the ELISPOT assay. Rotavirus-specific antibody-secreting cells were predominantly present in the alpha4beta7+ subpopulation. The flow cytometry assay we have described will permit future studies to characterize the phenotype of virus-specific B cells and could be useful in the study of the immunogenicity and protective efficacy of RV vaccines and the identification of markers of protective immunity.


Asunto(s)
Linfocitos B/inmunología , Integrinas/análisis , Infecciones por Rotavirus/inmunología , Rotavirus/inmunología , Adolescente , Adulto , Anciano , Anticuerpos Antivirales/análisis , Células Productoras de Anticuerpos/fisiología , Linfocitos B/química , Diarrea/inmunología , Humanos , Inmunofenotipificación , Lactante , Integrinas/fisiología , Persona de Mediana Edad , Receptores de Antígenos de Linfocitos B/análisis
15.
Microbes Infect ; 3(12): 971-84, 2001 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-11580984

RESUMEN

The determinants of the prevalence of CD8(+) T cells in the inflamed myocardium of Trypanosoma cruzi-infected patients and experimental animals are undefined. Using C3H/He mice infected with the Colombiana strain of T. cruzi, we found that the distribution of CD4(+)/CD8(-) and CD4(-)/CD8(+) T cells in the myocardium mirrors the frequency of cells expressing the CD62L(Low)LFA-1(High)VLA-4(High) activation phenotype among CD4(+)/CD8(-) and CD4(-)/CD8(+ )peripheral blood T cells. Consistently, vascular cell adhesion molecule-1-positive endothelial cells and a fine fibronectin network surrounding VLA-4(+) mononuclear cells were found in the inflamed myocardium. Further, interferon gamma (IFN-gamma) and IFN-gamma-induced chemokines (RANTES, MIG and CRG-2/IP-10), as well as JE/MCP-1 and MIP1-alpha, were found to be the dominant cytokines expressed in situ during acute and chronic myocarditis elicited by T. cruzi. In contrast, interleukin 4 mRNA was only detected during the chronic phase. Altogether, the results indicate that the distribution of T-cell subsets in the myocardium of T. cruzi-infected mice reflects the particular profile of adhesion molecules acquired by most peripheral CD8(+) T lymphocytes and point to the possibility that multiple IFN-gamma-inducible molecules present in the inflamed tissue contribute to the establishment and maintenance of T. cruzi-induced myocarditis.


Asunto(s)
Linfocitos T CD8-positivos/inmunología , Cardiomiopatía Chagásica/inmunología , Integrinas/análisis , Interferón gamma/farmacología , Selectina L/análisis , Antígeno-1 Asociado a Función de Linfocito/análisis , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Receptores Mensajeros de Linfocitos/análisis , Animales , Moléculas de Adhesión Celular/biosíntesis , Cardiomiopatía Chagásica/parasitología , Cardiomiopatía Chagásica/patología , Quimiocinas/biosíntesis , Citocinas/biosíntesis , Femenino , Inmunofenotipificación , Integrina alfa4beta1 , Ratones , Ratones Endogámicos C3H , Miocardio/patología , Parasitemia/mortalidad
16.
Histopathology ; 37(2): 118-23, 2000 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-10931234

RESUMEN

AIMS: Polymorphous low-grade adenocarcinoma (PLGA) and adenoid cystic carcinoma (ACC) are malignant salivary gland tumours bearing many similar histological patterns. This study was undertaken to show how the presence and distribution of collagen IV and laminin, and their ligands (integrin alpha2beta1 and alpha3beta1 components), can reveal histoarchitectural differences which distinguish these two entities. METHODS AND RESULTS: Five cases of ACC and five cases of PLGA from the archives of the Oral Pathology Department of the School of Dentistry of the São Paulo University were submitted to immunostaining with the antibodies to collagen IV, laminin, and integrins alpha2beta1 and alpha3beta1 using the streptavidin-biotin-peroxidase technique. Positive and negative controls were included. PLGA showed a thin line of collagen IV and laminin surrounding structures composed of a single cell layer. Integrins were expressed as a widespread and granular pattern. A thick line of collagen and laminin was observed around the neoplastic structures of ACC. Both integrins were expressed in intercellular spaces and around luminal spaces of tubular structures. CONCLUSIONS: Collagen IV and laminin, and their integrin ligands, are useful in demonstrating that neoplastic ductal units of PLGA are composed of a single cell layer, being distinct from ACC which contains structures composed of two layers of neoplastic cells.


Asunto(s)
Adenocarcinoma/metabolismo , Carcinoma Adenoide Quístico/metabolismo , Neoplasias de las Glándulas Salivales/metabolismo , Adenocarcinoma/patología , Carcinoma Adenoide Quístico/patología , Colágeno/análisis , Diagnóstico Diferencial , Humanos , Inmunohistoquímica , Integrina alfa3beta1 , Integrinas/análisis , Laminina/análisis , Receptores de Colágeno , Neoplasias de las Glándulas Salivales/patología
17.
Eur Respir J ; 9(10): 1995-2001, 1996 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-8902456

RESUMEN

Cigarette smoking produces peripheral airway inflammation in all smokers, and chronic airways obstruction in approximately 20% of heavy smokers. The present study was designed to test the hypothesis that airways obstruction is related to changes in the expression of adhesion molecules involved in the recruitment of cells to sites of inflammation in the lung. Freshly resected lungs from heavy smokers with airways obstruction (n = 10) and from heavy smokers with normal lung function (n = 10) were collected in the operating room, inflated with optimal cutting temperature (OCT) medium and frozen over liquid nitrogen. Six micrometres thick cryostat sections cut from random samples of this tissue were stained, using immunohistochemistry, with monoclonal antibodies to the adhesion molecules on leucocytes: L-selectin, very late activation antigen-4 (VLA-4), CD11a/CD18, CD11b/CD18, CD11c/CD18; and on endothelial and epithelial surfaces: E-selectin, P-selectin, vascular cell adhesion molecule (VCAM-1), intercellular adhesion molecule (ICAM)-1 and ICAM-2 using the alkaline phosphatase anti-alkaline phosphatase (APAAP) technique. The slides were coded and the expression of each molecule scored by three observers using a semiquantitative grading system. Two inducible adhesion molecules, E-selectin on endothelium and CD11b on leucocytes, were also evaluated using quantitative morphometric analysis. The results showed a distribution of adhesion molecules that was consistent with the inflammatory response in the airways and parenchyma of all subjects but failed to show any differences between those with or without airways obstruction. We conclude that development of airways obstruction in heavy smokers cannot be explained by differences in the expression of adhesion molecules known to be involved in the control of cell traffic in the lung.


Asunto(s)
Moléculas de Adhesión Celular/genética , Regulación de la Expresión Génica , Enfermedades Pulmonares Obstructivas/metabolismo , Fumar/metabolismo , Anciano , Anticuerpos Monoclonales , Antígenos CD/análisis , Antígenos CD/genética , Antígenos CD11/análisis , Antígenos CD11/genética , Antígenos CD18/análisis , Antígenos CD18/genética , Moléculas de Adhesión Celular/análisis , Selectina E/análisis , Selectina E/genética , Endotelio/metabolismo , Endotelio/patología , Epitelio/metabolismo , Epitelio/patología , Femenino , Humanos , Inmunohistoquímica , Integrina alfa4beta1 , Integrina beta1/análisis , Integrina beta1/genética , Integrinas/análisis , Integrinas/genética , Molécula 1 de Adhesión Intercelular/análisis , Molécula 1 de Adhesión Intercelular/genética , Selectina L/análisis , Selectina L/genética , Leucocitos/metabolismo , Leucocitos/patología , Pulmón/metabolismo , Pulmón/patología , Enfermedades Pulmonares Obstructivas/genética , Enfermedades Pulmonares Obstructivas/patología , Masculino , Persona de Mediana Edad , Selectina-P/análisis , Selectina-P/genética , Neumonía/genética , Neumonía/metabolismo , Neumonía/patología , Receptores Mensajeros de Linfocitos/análisis , Receptores Mensajeros de Linfocitos/genética , Receptores de Antígeno muy Tardío/análisis , Receptores de Antígeno muy Tardío/genética , Humo , Fumar/genética , Fumar/patología , Molécula 1 de Adhesión Celular Vascular/análisis , Molécula 1 de Adhesión Celular Vascular/genética
18.
J Pediatr ; 126(4): 530-6, 1995 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-7535352

RESUMEN

OBJECTIVE: To evaluate the cell-mediated immune status of children with recurrent respiratory tract infections. DESIGN: We evaluated the cell-mediated immune status of 76 patients referred because of recurrent infection. Patients were divided into those with serologic abnormalities and those without such findings. Twenty-three healthy children served as control subjects. Studies of lymphocyte phenotype included CD4+ CD29+ cells (an immunologically mature phenotype), lymphocyte proliferation studies, cytokine production including interleukin-2 (IL-2), IL-4, IL-6, and interferon gamma), and measurement of in vitro IgM and IgG synthesis. RESULTS: Lymphocyte proliferation and T-cell phenotype were similar in both patient groups as well as in control subjects. The proportions of CD4+ CD29+ cells at different ages were similar in all groups. Patients with serologic abnormalities (e.g., partial IgA deficiency, partial IgG subclass deficiency) produced more IL-2 and IL-4 than did other patients. The control population had greater spontaneous IgM and IgG synthesis than the patient groups. CONCLUSION: Routine studies of T-cell function of patients with recurrent infection provide little information useful in making clinical decisions.


Asunto(s)
Antígenos CD/análisis , Antígenos CD4/análisis , Citocinas/sangre , Inmunidad Celular , Integrinas/análisis , Infecciones del Sistema Respiratorio/inmunología , Antígenos Bacterianos/biosíntesis , Vacunas Bacterianas/inmunología , Niño , Preescolar , Humanos , Inmunidad Celular/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Inmunofenotipificación , Lactante , Integrina beta1 , Interferón gamma/sangre , Interleucina-2/sangre , Interleucina-4/sangre , Interleucina-6/sangre , Activación de Linfocitos , Análisis por Apareamiento , Vacunas Neumococicas , Recurrencia
19.
Trans R Soc Trop Med Hyg ; 88(5): 596-9, 1994.
Artículo en Inglés | MEDLINE | ID: mdl-7992351

RESUMEN

Human invasive amoebiasis is highly destructive, causing rapid necrosis and liquefaction of all tissues reached by the trophozoites. Degradation of extracellular matrix components (EMC) has been demonstrated during invasion of the basal lamina. Pursuing the idea that trophozoites might behave similarly to other invasive cells with respect to their interaction with EMC, plasma membrane proteins biochemically or functionally related to integrins were looked for. A 140 kDa molecular mass membrane protein from Entamoeba histolytica trophozoites with the characteristics of a beta 1 integrin-like fibronectin receptor was identified.


Asunto(s)
Entamoeba histolytica/química , Integrinas/análisis , Animales , Adhesión Celular , Técnica del Anticuerpo Fluorescente , Peso Molecular , Receptores de Fibronectina/análisis
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