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Lymphocyte Blastogenesis Studied by Volume Spectroscopy
Author(s) -
STEEN H. B.,
NIELSEN V.
Publication year - 1979
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1979.tb03268.x
Subject(s) - concanavalin a , thymidine , incubation , volume (thermodynamics) , coulter counter , microbiology and biotechnology , mitosis , lymphocyte , chemistry , biology , immunology , andrology , in vitro , biochemistry , medicine , physics , quantum mechanics
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 wan dispersed by 3 h of incubation will α‐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 μg/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 3 H‐thymidine incorporation could he 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 (150μg/ml) which strongly suppressed 3 H‐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.