Fatty acid metabolism in human lymphocytes. II. Activation of fatty acid desaturase-elongase systems during blastic transformation.
The fatty acid desaturation-elongation ability of human T-lymphocytes during blastic transformation was determined both by gas-liquid chromatography and incubation with radiolabeled precursors. Human peripheral blood mononuclear cells (PBMC) were activated with phytohemagglutinin (PHA) and cultured in media supplemented with different fatty acids (18:0, 18:1(n - 9), 18:2(n - 6), 18:3(n - 3) and 20:4(n - 6)) at a final concentration of 30 microM. All the fatty acids added were elongated by activated PBMC and the maximal activity was observed on 20:4(n - 6) (a 25% of conversion to 22:4(n - 6)). Supplementation with stearic acid increased the proportion of oleic (from 21.4% to 23.7%) and eicosaenoic (from 3.1% to 5.7%) acids in cellular lipids, indicating the existence of a delta 9-desaturase activity. Supplementation with linoleic and linoleic acids increased slightly the cell content in their more unsaturated derivatives. Direct measurement of desaturase activities was performed by incubating quiescent and activated PBMC with [1-14C]stearic, [1-14C]linoleic and [1-14C]linolenic acids. Quiescent cells exhibited a very low delta 9-desaturase and no sign of delta 6-desaturase activity. A moderate and progressive activation of delta 9-, delta 6- and delta 5-desaturases was observed during blastic transformation of human PBMC. Up to 8% of 18:0 was converted to monoenes, 4% and 1.5% of 18:2(n - 6) was converted to trienes and tetraenes, respectively, and 14.5% of 18:3(n - 3) was converted to pentaenes. The maximal relative activities were found after 48 h of PHA-stimulation for delta 9-desaturase (around 90 pmol of 18:0 converted per 10(6) cells in the last 24 h) and at 72 h for delta 6- and delta 5-desaturases (around 75 and 140 pmol of 18:2 and 18:3, respectively, converted per 10(7) cells in the last 24 h). Although these activities are not enough to explain all the changes in fatty acid composition of human PBMC during blastic transformation, they may contribute to a more controlled cell phospholipid composition.
Anel
A
A
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Zaragoza, Spain.
Naval
J
J
González
B
B
Uriel
J
J
Piñeiro
A
A
D000123:Acetyltransferases / Q000097:blood*; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000097:blood / Q000494:pharmacology; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D044943:Fatty Acid Desaturases / Q000097:blood*; D000081031:Fatty Acid Elongases; D005227:Fatty Acids / Q000097:blood* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D019787:Linoleic Acid; D008041:Linoleic Acids / Q000097:blood / Q000494:pharmacology; D008042:Linolenic Acids / Q000097:blood / Q000494:pharmacology; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects / Q000201:enzymology*; D019301:Oleic Acid; D009829:Oleic Acids / Q000097:blood / Q000494:pharmacology; D010835:Phytohemagglutinins / Q000494:pharmacology; D013229:Stearic Acids / Q000097:blood / Q000494:pharmacology
D001095:Arachidonic Acids; D005227:Fatty Acids; D008041:Linoleic Acids; D008042:Linolenic Acids; D009829:Oleic Acids; D010835:Phytohemagglutinins; D013229:Stearic Acids; D016718:Arachidonic Acid; D019301:Oleic Acid; C031183:stearic acid; D019787:Linoleic Acid; D044943:Fatty Acid Desaturases; D000123:Acetyltransferases; D000081031:Fatty Acid Elongases