Cell kinetic effects of incorporated /sup 3/H-thymidine on proliferating human lymphocytes: flow cytometric analysis using the DNA/nuclear protein method

Journal Article · Sun Sep 01 04:00:00 EDT 1985 · Cytometry (Baltimore); (United States)

OSTI ID:6380658

Pollack, A; Moulis, H; Greenstein, D B; Block, N L; Irvin, G.L. 3d.

Phytohemagglutinin-stimulated human peripheral blood lymphocytes incorporating high concentrations of /sup 3/H-thymidine accumulate in G2 and show a consequent reduction in the number of cells entering M (division delay). The simultaneous flow cytometric analysis of DNA content (propidium iodide fluorescence) and nuclear protein content (fluorescein isothiocyanate fluorescence) allows for the accurate quantitation of these events; G2 and M are separated in the bivariate distributions. A good correlation was observed between mitotic indices, quantitated by manually counting mitotic cells, and integration of the M area in DNA/nuclear protein histograms. Moreover, significant differences in G2 nuclear protein levels were found between untreated and /sup 3/H-thymidine-treated lymphocytes. In order to characterize this effect, G2 was empirically divided into low nuclear protein (G2A) and high nuclear protein (G2B) compartments. /sup 3/H-thymidine caused an initial accumulation of lymphocytes in G2A, followed within 3-6 h by a gradual movement of some cells into G2B, with a subsequent accumulation of cells in G2B. The results suggest that the distribution of cells in G2 (G2A and G2B), the average nuclear protein content of G2B cells, and the proportion of cells in M are parameters that when used in combination provide a unique description of radiobiological effects.

• Research Organization: Veterans Administration Medical Center, Miami, FL
• OSTI ID: 6380658
• Journal Information: Cytometry (Baltimore); (United States), Journal Name: Cytometry (Baltimore); (United States) Vol. 5; ISSN CYTOD
• Country of Publication: United States
• Language: English

Related Subjects

550401* -- Genetics-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AGGLUTININS
ANIMAL CELLS
ANTIBODIES
AZINES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CONSTITUENTS
CELL NUCLEI
CELL PROLIFERATION
CONNECTIVE TISSUE CELLS
DNA
FLUORESCENCE
HEMAGGLUTININS
HETEROCYCLIC COMPOUNDS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LEUKOCYTES
LUMINESCENCE
LYMPHOCYTES
MATERIALS
METABOLISM
MITOTIC INDEX
NUCLEIC ACIDS
NUCLEOPROTEINS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PROTEINS
PYRIMIDINES
REACTION KINETICS
RIBOSIDES
SOMATIC CELLS
THYMIDINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS