RESUMEN
Vascular endothelial cells recognize blood-borne circulating cells and allow them to extravasate in a tissue-specific manner. Because this property determines the selectivity of lymphocyte homing, it is fundamental in physiological as well as pathological processes (inflammation, autoimmune diseases, metastasis). As a tool to assess the molecular basis of endothelium selectivity, microvascular endothelial cell lines of distinct tissue origin were established. Endothelial cells, isolated from lymphoid tissues (lymph nodes and appendix) and from nonlymphoid immune sites--intestine, lung, and skin--were immortalized in vitro. Their general endothelial characteristics, such as the presence of von Willebrand factor (wWf), angiotensin-converting enzyme (ACE), VE-cadherin, and the intracellular E-selectin, were preserved. This article shows that these cell lines display phenotypic characteristics related to their tissue origin. Hence, endothelial cells from lymph nodes expressed peripheral lymph node addressins (PNAds). Endothelial cells from nonlymphoid tissues were ICAM-1 (intercellular adhesion molecule-1) and CD49e positive, whereas P-selectin was not equally distributed among the cell lines. Endothelial cells from mucosal sites reacted with antibody against human MAdCAM-1 (mucosal addressin cell adhesion molecule). In the adhesion test, lymphoid and myeloid cells adhere to endothelial cell lines in a distinct manner. These lines could be useful to study molecular mechanisms involved in tissue-specific cell-cell interaction.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Línea Celular Transformada/metabolismo , Linaje de la Célula/fisiología , Quimiotaxis de Leucocito/fisiología , Endotelio Vascular/metabolismo , Activación de Linfocitos/fisiología , Animales , Antígenos CD , Cadherinas/metabolismo , Adhesión Celular/fisiología , Comunicación Celular/fisiología , Línea Celular Transformada/citología , Cricetinae , Selectina E/metabolismo , Endotelio Vascular/citología , Humanos , Inmunoglobulinas/metabolismo , Integrina alfa5/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/citología , Intestinos/inmunología , Tejido Linfoide/citología , Tejido Linfoide/inmunología , Tejido Linfoide/metabolismo , Mucoproteínas/metabolismo , Selectina-P/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Fenotipo , Piel/citología , Piel/inmunología , Piel/metabolismo , Factor de von Willebrand/metabolismoRESUMEN
FGF-1 stimulated DNA synthesis and induced expression of IL-2 receptors in the murine fibrosarcoma cell line, F69-3. Concomitant treatment with IL-2 abolished the stimulation of DNA synthesis, but not binding of FGF-1 to the FGF-receptors or subsequent endocytosis of the bound growth factor. Also, it did not inhibit activation of the FGF-receptor tyrosine kinase or stimulation of the downstream effector, MAP kinase. Treatment with IL-2 prevented transport of FGF-1 to the nuclear fraction in a time- and dose-dependent manner that parallelled the inhibition of FGF-1 stimulated DNA synthesis. The data support our earlier finding that transport of FGF-1 to the nucleus is an important event in the mechanism of stimulation of DNA synthesis induced by the growth factor, and they demonstrate that treatment with a cytokine can modulate the cellular response to FGF-1.
Asunto(s)
Factor 2 de Crecimiento de Fibroblastos/farmacología , Fibrosarcoma/metabolismo , Interleucina-2/farmacología , Animales , Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Núcleo Celular/metabolismo , ADN de Neoplasias/biosíntesis , Activación Enzimática/efectos de los fármacos , Factor 1 de Crecimiento de Fibroblastos , Factor 2 de Crecimiento de Fibroblastos/antagonistas & inhibidores , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Unión Proteica/efectos de los fármacos , Receptores de Factores de Crecimiento de Fibroblastos/metabolismo , Transducción de Señal/efectos de los fármacos , Células Tumorales CultivadasRESUMEN
Murine interleukin 2 (mIL-2) cDNA was introduced through lipofection into cells of murine F-69-3 fibrosarcoma line established in vitro from tumors induced chemically in athymic mice. Using a modified MTT bioassay in CTLL-2 indicator line the F-69-3/IL-2 transfectants were found to secrete between 650-1750 laboratory units (LU) of IL-2/5 x 10(5) cells/48 h in restricted culture conditions. When inoculated subcutaneously to immunocompetent BALB/c or CD2F1 mice, the transfected cells showed reduced tumorigenic potential as compared with parental F-69-3/wt or control F-69-3/neo cells. The rejection of F-69-3/IL-2 tumors required an intact immune system as they grew progressively in athymic mice. The majority of immunocompetent mice that rejected IL-2 secretors were found to be protected against subsequent challenge with parental cells. These preliminary results suggest that IL-2-transfected murine fibrosarcoma could be used as a model for studying mechanisms underlying the antitumor immune response.