Lymphocyte blastogenesis studied by volume spectroscopy.
The blastogenesis of human blood lymphocytes has been studied by measuring the distribution of cellular and nuclear volumes in concanavalin A (Con A)-stimulated leucocyte cultures by means of a modified Coulter counter. Agglutinated and adherent cells were dispersed by 3 h of incubation with alpha-methyl-mannoside and EDTA. Fresh suspensions contained 5--35% monocytes with an average cellular volume 2--2.5 times that of small lymphocytes. The nuclear volume of monocytes was similar to that of small lymphocytes within 10% and remained constant upon stimulation. In cultures given the optimal dose of Con A, 30 micrograms/ml, measurable cell growth commenced within 8--12 h, whereas nuclear growth began some 6 h later, which is approximately 6--8 h before increased 3H-thymidine incorporation could be detected. The average volume of responding cells grew by a factor of about 4 until mitosis, which started after around 48 h, whereas the average nuclear volume increased by a factor of about 3. The percentage of cells responding by volume growth increased with mitogen concentration even for hyperoptimal concentrations (150 micrograms/ml) which strongly suppressed 3H-thymidine incorporation. The results exemplify the type of information obtained by volume spectroscopy of leucocyte cultures. This method appears to have several advantages over conventional assays of immune response.
D002467:Cell Nucleus; D002470:Cell Survival; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology; D003703:Demecolcine / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D007958:Leukocyte Count; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000166:cytology*; D008362:Mannosides / Q000494:pharmacology; D013057:Spectrum Analysis
D008362:Mannosides; D003208:Concanavalin A; D004492:Edetic Acid; D003703:Demecolcine