Cyclic nucleotide phosphodiesterases from purified human CD4+ and CD8+ T lymphocytes.
BACKGROUND
CD4+ and CD8+ T-lymphocytes are suggested to differentially affect airway inflammation in asthma. Agents which increase intracellular cAMP levels, such as PDE inhibitors, have been shown to diminish lymphocyte growth and differentiation, and to affect cytokine expression. Differences in the PDE isoenzyme profile between CD4+ and CD8+ cells might form a basis to differentially modify their functions by PDE inhibitors.


OBJECTIVE
The study investigates and compares the PDE isoenzyme activity profiles of human peripheral blood CD4+ and CD8+ T-lymphocytes.


METHODS
CD4+ and CD8+ T-lymphocytes were purified (> 98%) from peripheral blood mononuclear cells by negative selection. PDE isoenzyme activity profiles were investigated using PDE isoenzyme selective inhibitors and activators.


RESULTS
In CD4+ and CD8+ T-lymphocyte homogenates, PDE IV and PDE III activities were the predominant PDE isoenzyme activities at 0.5 microM cyclic nucleotide substrate concentrations. PDE IV was localized in the soluble fraction whereas PDE III was membrane bound. Low PDE I, II and V activities were detected. About 20% of total cAMP hydrolysing capacity at 0.5 microM cAMP was insensitive to PDE isoenzyme selective inhibitors and activators and therefore could not be assigned to PDE I-IV. The PDE isoenzyme pattern was not different between CD4+ and CD8+ T-lymphocytes. Moreover, representative inhibitors of PDE III and IV activity inhibited cAMP hydrolysis in soluble fractions of both T-lymphocyte subsets with similar potency. Enzyme kinetic analysis similarly did not reveal differences between CD4+ and CD8+ T-lymphocytes.


CONCLUSIONS
Normal CD4+ and CD8+ T-lymphocytes are likely to be equally sensitive targets for the effects of PDE inhibitors.
Tenor
H
H
Immunopharmacology Group, University of Southampton, UK.
Staniciu
L
L
Schudt
C
C
Hatzelmann
A
A
Wendel
A
A
Djukanović
R
R
Church
M K
MK
Shute
J K
JK
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors / Q000097:blood; D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000097:blood; D015496:CD4-Positive T-Lymphocytes / Q000201:enzymology*; D018414:CD8-Positive T-Lymphocytes / Q000201:enzymology*; D002469:Cell Separation; D006801:Humans; D007527:Isoenzymes / Q000097:blood; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000097:blood*; D013347:Subcellular Fractions / Q000201:enzymology
D007527:Isoenzymes; D010726:Phosphodiesterase Inhibitors; D010727:Phosphoric Diester Hydrolases; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D015106:3',5'-Cyclic-GMP Phosphodiesterases