Differential signal transduction pathways regulating interleukin-2 synthesis and interleukin-2 receptor expression in stimulated human lymphocytes.
In human peripheral blood lymphocytes stimulated via the T-cell antigen receptor/CD3 complex IL-2 synthesis and cellular proliferation were effectively inhibited by a concentration of ouabain as low as 50 nM, whilst the expression of high affinity IL-2 receptors was not influenced. Binding of the monoclonal antibody, BMA 031 to the T-cell antigen receptor/CD3 complex resulted in a bimodal activation of protein kinase C. The activation of protein kinase C-alpha in the early phase of T-lymphocyte activation was not affected by 50 nM ouabain, in contrast sustained activation of protein kinase C-beta, between 90-240 min of stimulation was completely abolished by the cardiac glycoside. When protein kinase C was directly activated by PMA + ionomycin, 50 nM ouabain was ineffective in inhibiting protein kinase C activation, as well as subsequent IL-2 synthesis, suggesting that the glycoside interfered with signal transducing mechanism(s) upstream of the activation of protein kinase C. Ouabain had no influence on the elevation of intracellular calcium concentration in BMA 031 stimulated lymphocytes, ruling out the possibility that it interfered with the T-cell antigen receptor dependent phosphatidylinositol response. In contrast, lysophosphatide acyltransferase catalysed elevated incorporation of polyunsaturated fatty acids was effectively inhibited by low concentrations of ouabain in BMA 031-stimulated T-lymphocytes, whereas stimulation with PMA + ionomycin had no influence on the plasma membrane phospholipid fatty acid metabolism. These results suggest, that differential signal transduction pathways are involved in the activation of protein kinases C-alpha and -beta. They implicate that elevated incorporation of polyunsaturated fatty acids into plasma membrane phospholipids might contribute to sustained activation of protein kinase C-beta, and establish a link between activation of protein kinase C-beta and induction of IL-2 synthesis in human lymphocytes.
Szamel
M
M
Department of Pharmacology and Toxicology, Medical School Hannover, Germany.
Leufgen
H
H
Kurrle
R
R
Resch
K
K
D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004789:Enzyme Activation; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis*; D015759:Ionomycin / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism; D010042:Ouabain / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000276:immunology; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis*; D015398:Signal Transduction
D005231:Fatty Acids, Unsaturated; D007376:Interleukin-2; D007527:Isoenzymes; D008563:Membrane Lipids; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D015375:Receptors, Interleukin-2; D015759:Ionomycin; D010042:Ouabain; C496628:PRKCA protein, human; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D002118:Calcium