Expression of glycolytic isoenzymes in activated human peripheral lymphocytes: cell cycle analysis using flow cytometry.
Human peripheral lymphocytes activated with concanavalin A and phorbol myristate ester exhibit an increase in glycolysis on a time-course similar to that for DNA synthesis. Elevated glycolysis is accompanied by increased specific activities of the glycolytic enzymes. Increased enzyme activities are accounted for by the appearance of specific isoenzyme forms (muscle forms) normally expressed in rapidly growing tumor cells or in growth-stimulated cells. In the present study we analyzed the expression of the glycolytic isoenzymes during cell cycle progression of activated human lymphocytes using two-parameter (DNA and protein) flow cytometry. Time-course studies and analysis of subpopulations prepared by elutriation centrifugation showed that the inducible isoenzymes are expressed at low levels or not at all in G0 cells. They are expressed first during the G0 to G1 transition or in early G1. However, expression increases throughout G1, reaching a maximum in S-phase. Thus, induction of glycolytic isoenzymes provides an excellent marker of T-cell activation and progression toward DNA synthesis.
Marjanovic
S
S
Department of Biochemistry, Arrhenius Laboratory, University of Stockholm, Sweden.
Skog
S
S
Heiden
T
T
Tribukait
B
B
Nelson
B D
BD
D002453:Cell Cycle; D003208:Concanavalin A / Q000494:pharmacology; D005434:Flow Cytometry; D006019:Glycolysis; D006801:Humans; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000201:enzymology* / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors
D007527:Isoenzymes; D007773:Lactates; D003208:Concanavalin A; D007770:L-Lactate Dehydrogenase; D011770:Pyruvate Kinase; D010751:Phosphopyruvate Hydratase; D013755:Tetradecanoylphorbol Acetate