Asunto(s)
Apoptosis/fisiología , Proteínas Portadoras/metabolismo , Células Eucariotas/metabolismo , Péptidos y Proteínas de Señalización Intracelular , Receptores de Factor de Crecimiento Nervioso/metabolismo , Receptores del Factor de Necrosis Tumoral/metabolismo , Secuencia de Aminoácidos/genética , Animales , Proteínas Reguladoras de la Apoptosis , Células COS , Proteínas Portadoras/genética , Citoplasma/metabolismo , Proteína Relacionada con TNFR Inducida por Glucocorticoide , Ratones , Datos de Secuencia Molecular , Mutación/genética , Unión Proteica/fisiología , Estructura Terciaria de Proteína/fisiología , Receptores de Factor de Crecimiento Nervioso/genética , Receptores OX40 , Receptores del Factor de Necrosis Tumoral/genética , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/genética , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/metabolismoRESUMEN
To investigate the role of major histocompatibility complex class I and bone marrow stromal cells on in vitro differentiation of natural killer cells, a CD44(low/-) CD2- population was isolated from mouse bone marrow. This NK-1.1- CD3- LFA-3+ B220+ population, when stimulated with IL-2 and co-cultured with supportive syngeneic stromal cells, generated populations of NK-1.1+ Ly49A+ Ly49C/I+ CD3- mature natural killer cells. The effect of anti-H-2b monoclonal antibodies (mAbs) on this phenomenon was assayed. Pre-adhesion of anti-H-2b mAbs to the stromal cells did not exert any effect, whereas when the same mAbs were pre-adhered to progenitors, there was a inhibition of natural killer cell generation that was maximum when the mAbs were added directly to cultures. In addition, the anti-H-2b mAbs did not inhibit the IL-2-induced proliferation of mature natural killer cells. Allogeneic but not H-2b-deficient stromal cells decreased the expression of Ly-49C/I but not Ly49A, thus suggesting that stromal cell haplotypes qualitatively influence the expression of Ly49s repertoire.
Asunto(s)
Antígenos Ly , Antineoplásicos/farmacología , Antígenos H-2/inmunología , Interleucina-2/farmacología , Células Asesinas Naturales/fisiología , Glicoproteínas de Membrana/inmunología , Animales , Células de la Médula Ósea , Técnicas de Cultivo de Célula , Diferenciación Celular , División Celular , Regulación de la Expresión Génica , Lectinas Tipo C , Ratones , Ratones Endogámicos C57BL , Subfamilia A de Receptores Similares a Lectina de Células NK , Receptores Similares a Lectina de Células NK , Células del Estroma/inmunologíaAsunto(s)
Empalme Alternativo , Variación Genética , ARN Mensajero/genética , Receptores de Factor de Crecimiento Nervioso/genética , Receptores del Factor de Necrosis Tumoral/genética , Secuencia de Aminoácidos , Exones , Proteína Relacionada con TNFR Inducida por Glucocorticoide , Humanos , Intrones , Datos de Secuencia Molecular , Señales de Clasificación de Proteína/química , Receptores de Factor de Crecimiento Nervioso/química , Receptores del Factor de Necrosis Tumoral/química , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T/metabolismoRESUMEN
Long-term bone marrow cultures were used to investigate the effect of IL-2, a cytokine widely used in immunotherapy, on natural killer cell differentiation. Specifically, the role of MHC was evaluated by comparing normal B6 and class I-deficient TAP-1-/- mice. The number of cells generated after a 13-day culture was the same in cell cultures from TAP-1-/- or B6 mice but the relative number of natural killer cells, identified as NK-1.1+CD3- cells by flow cytometry analysis, was increased in TAP-1-/- compared to B6 cultures (74.4% and 63.9%, respectively). Addition of an anti-class I mAb determined a strong inhibition of natural killer cell generation in B6 cultures, and its effect was specific since no effect was seen in TAP-1-/- cell cultures. TAP-1-/- natural killer cells or the few natural killer cells escaping the inhibitory effect of anti-class I mAb, were less cytotoxic than total B6 natural killer cells against target cell lines of different haplotype.