The generation of 1H-NMR-detectable mobile lipid in stimulated lymphocytes: relationship to cellular activation, the cell cycle, and phosphatidylcholine-specific phospholipase C.
Mobile lipids detected using 1H-NMR in stimulated lymphocytes were correlated with cell cycle phase, expression of the interleukin-2 receptor alpha and proliferation to assess the activation status of the lymphocytes. Mobile lipid levels, IL-2R alpha expression and proliferation increased after treatment with PMA and ionomycin. PMA or ionomycin stimulation alone induced increased IL-2R alpha expression but not proliferation. PMA- but not ionomycin-stimulation generated mobile lipid. Treatment with anti-CD3 antibody did not increase IL-2R alpha expression or proliferation but did generate increased amounts of mobile lipid. The cell cycle status of thymocytes treated with anti-CD3, PMA or ionomycin alone indicated an accumulation of the cells in the G1 phase of the cell cycle. The generation of mobile lipid was abrogated in anti-CD3 antibody-stimulated thymic lymphocytes but not in splenic lymphocytes, using a phosphatidylcholine-specific phospholipase C (PC-PLC) inhibitor which blocked cells in the G1/S phase of the cell cycle. This suggests that the 1H-NMR-detectable mobile lipid may be generated in anti-CD3 antibody-stimulated thymic lymphocytes by the action of PC-PLC activity via the catabolism of PC, in the absence of classical signs of activation.
Veale
M F
MF
Department of Pathology, University of Sydney, New South Wales, Australia.
Roberts
N J
NJ
King
G F
GF
King
N J
NJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001952:Bridged-Ring Compounds / Q000494:pharmacology; D017252:CD3 Complex / Q000276:immunology; D002453:Cell Cycle / Q000276:immunology*; D002455:Cell Division / Q000187:drug effects / Q000276:immunology; D002470:Cell Survival / Q000187:drug effects / Q000276:immunology; D004338:Drug Combinations; D015759:Ionomycin / Q000494:pharmacology; D050356:Lipid Metabolism; D008055:Lipids / Q000096:biosynthesis*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008810:Mice, Inbred C57BL; D009636:Norbornanes; D010713:Phosphatidylcholines / Q000378:metabolism*; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D013601:T-Lymphocytes / Q000201:enzymology / Q000276:immunology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013859:Thiocarbamates; D013871:Thiones / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D001952:Bridged-Ring Compounds; D017252:CD3 Complex; D004338:Drug Combinations; D008055:Lipids; D009636:Norbornanes; D010713:Phosphatidylcholines; D015375:Receptors, Interleukin-2; D013859:Thiocarbamates; D013871:Thiones; D015759:Ionomycin; C046498:tricyclodecane-9-yl-xanthogenate; D010738:Type C Phospholipases; C105598:phosphatidylcholine-specific phospholipase C; D013755:Tetradecanoylphorbol Acetate