RESUMEN
T cell subset determinations were performed on 146 peripheral blood samples from healthy volunteers, and on 112 samples from immune deficient patients using two fluorescence-activated cell sorters (the FACSIV laser, and the FACSTm mercury lamp analyzer). The procedures necessary for the use and calibration of the FACSTm analyzer are discussed, and detailed. Using the FACSTm analyzer, counts were made of T and B cell subsets in 28 patients with multiple infections, 9 patients suffering from the acquired immune deficiency syndrome (AIDS) and 16 patients with a primary immunodeficiency disease. These results were compared with data obtained from 47 healthy volunteers, as control references. Results from the two instruments proved closely comparable, both qualitatively and quantitatively.
Asunto(s)
Linfocitos B/clasificación , Separación Celular , Citometría de Flujo , Linfocitos T/clasificación , Fluoresceína-5-Isotiocianato , Fluoresceínas , Humanos , Rayos Láser , Mercurio , TiocianatosRESUMEN
Optimal conditions are defined for hybridoma formation between mouse spleen cells and mouse myeloma cells. The results of using different numbers of spleen cells in the fusion process is reported in 2 parts. Part I deals with the number of spleen cells in relation to hybridoma formation and antibody production. Part II treats of the purity of hybridoma clones and the loss of antibody production following fusion. Part I. Two series of experiments show that when cell fusion is performed properly the total number of antibody producing clones is greater than in non-standard conditions. The yield of hybridomas obtained with a ratio of mouse myeloma to mouse spleen cells of 1:10 did not differ from that reported by De Blas et al. (1981). The number of hybridomas formed seems to depend mainly on the number of mouse spleen cells available. The most satisfactory yield of monoclonal antibodies is obtained under conditions producing growth in approximately 100% of the wells. Part II. Two weeks after fusion a number of antibody producing clones were cultured in limiting dilution. Analysis of the hybridomas indicated that at least 40% of the antibody producing clones disappear during the first 3 weeks. Antibody producing hybridomas were as a rule not outgrown by non-antibody producing clones.
Asunto(s)
Fusión Celular , Separación Celular/métodos , Hibridomas/inmunología , Activación de Linfocitos , Animales , Células Productoras de Anticuerpos/inmunología , Recuento de Células , Células Clonales/inmunología , Humanos , Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Mieloma Múltiple/inmunología , Bazo/citologíaAsunto(s)
Glicoproteínas , Mescalina/farmacología , Mitosis/efectos de los fármacos , Tubulina (Proteína) , Sitios de Unión , Biotransformación , Células Cultivadas , Glicoproteínas/metabolismo , Mescalina/metabolismo , Microtúbulos/metabolismo , Organoides/efectos de los fármacos , Relación Estructura-Actividad , Tubulina (Proteína)/metabolismoRESUMEN
HeLa cells were grown as monolayer cultures in Spinner medium and given 12-minute pulse labels of (3)H thymidine. Synchronous crops of cells labelled in the last minutes of S showed a metaphase pattern of chromosomes hot-labelled close to the kinetochores (centromeres). Such cells in subsequent stages of interphase showed a concentration of label at the nuclear envelope. A control experiment with cells labelled in the first minutes of S yielded chromosomes labelled at the tips and in the nucleolar organizers. These cells showed label concentrated in the central area of the nucleus in the next interphase. The results are compatible with the view that the kinetochore regions of chromosomes are attached to the nuclear envelope in interphase. The significance of these results is considered with reference to interpreting DNA replication patterns, the segregation of chromosomes, eukaryote differentiation and repression, and the rationale for chromosome numbers. It is argued that the nuclear envelope serves as a 'repressor organelle' in higher organisms, and that DNA methylation may be involved in the control of RNA transcription, and hence gene expression.