Regulatory roles of adenylate cyclase and cyclic nucleotide phosphodiesterases 1 and 4 in interleukin-13 production by activated human T cells.
We studied the activities of 3',5'-adenosine-cyclic monophosphate (cAMP)- synthesizing adenylate cyclase (AC) and cAMP-hydrolyzing cyclic nucleotide phosphodiesterase (PDE) in phytohemagglutinin (PHA)- or anti-CD3 plus anti-CD28-stimulated human T cells, and examined their roles in interleukin-13 (IL-13) production. The AC inhibitor MDL 12,330A [cis-N-(2-phenylcyclopentyl)azacyclotridec-1-en-2-amine hydrochloride] completely blocked PHA- or anti-CD3/CD28-induced IL-13 production. The PDE 1 inhibitor 8-methoxymethyl-3-isobutyl-1-methylxanthine or the PDE4 inhibitor rolipram partially inhibited IL-13 production, and the addition of both resulted in 100 or 85% inhibition in PHA- or anti-CD3/CD28-stimulated T cells, respectively. AC in T cells was transiently activated 5 min after stimuli, followed by the transient activation of PDE4 at 30 min. PDE1 activity, undetectable in resting T cells, was detected 3 hr after stimuli, and then increased gradually. Although PDE1-, 2-, 3-, and 4-independent PDE activity was low (< or = 15% of total), it began to increase 3 hr after anti-CD3/CD28; the increase was blocked by PDE7 antisense oligonucleotide, and such an increase was not induced by PHA. PHA or anti-CD3/CD28 induced PDE1B mRNA expression, undetectable in resting T cells. PDE4 mRNA level in T cells was not altered by either stimulus. PDE7 mRNA expression was detected in resting T cells, and was enhanced by anti-CD3/CD28, but not by PHA. The cAMP level of T cells increased 5 min after stimuli, returned to the basal level at 2 hr, and then continued to decrease. These results suggest that PHA or anti-CD3/CD28 initially (< or = 5 min) increases cAMP in T cells via AC, then reverses the increase via PDE4 (< or = 2 hr), and in the later phase (> 2 hr) further decreases cAMP via PDE1. Both the time-dependent increase and decrease of cAMP may be required for IL-13 production.
Kanda
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Department of Dermatology, Teikyo University, School of Medicine, 11-1, Kaga 2, Itabashi-Ku, Tokyo 173-8605, Japan. nmk@med.teikyo-u.ac.jp
Watanabe
S
S
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000328:Adult; D000906:Antibodies / Q000494:pharmacology; D018106:CD28 Antigens / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D000242:Cyclic AMP / Q000378:metabolism; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D018793:Interleukin-13 / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D008213:Lymphocyte Activation / Q000502:physiology*; D008297:Male; D010835:Phytohemagglutinins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D013997:Time Factors
D000067956:Adenylyl Cyclase Inhibitors; D000906:Antibodies; D018106:CD28 Antigens; D017252:CD3 Complex; D004791:Enzyme Inhibitors; D018793:Interleukin-13; D007527:Isoenzymes; D010835:Phytohemagglutinins; D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D000262:Adenylyl Cyclases