Inhibition of leukocyte function and interleukin-2 gene expression by 2-methylarachidonyl-(2'-fluoroethyl)amide, a stable congener of the endogenous cannabinoid receptor ligand anandamide.
Arachidonylethanolamide (anandamide, AEA) has been identified as an endogenous ligand for cannabinoid receptors CB1 and CB2. Characterization of the direct cannabimimetic actions of anandamide has been hampered by its short duration of action and rapid degradation in in vivo and in vitro systems to arachidonic acid, a precursor in the biosynthesis of a broad range of biologically active molecules. In the present studies, we utilized 2-methylarachidonyl-(2'-fluoroethyl)amide (F-Me-AEA), an analog of anandamide resistant to enzymatic degradation, to determine whether F-Me-AEA modulated T cell function similar to that of plant-derived cannabinoids. Indeed, F-Me-AEA at low micromolar concentrations exhibited a marked inhibition of phorbol ester plus calcium ionophore (PMA/Io)-induced IL-2 protein secretion and steady state mRNA expression. Likewise, a modest suppression of the mixed lymphocyte response was observed in the presence of F-Me-AEA indicating an alteration in T cell responsiveness to allogeneic MHC class II antigens. F-Me-AEA was also found to modestly inhibit forskolin-stimulated adenylate cyclase activity in thymocytes and splenocytes, a hallmark of cannabinoid receptor agonists. Further characterization of the influence of F-Me-AEA on the cAMP signaling cascade revealed an inhibition of CREB-1/ATF-1 phosphorylation and subsequently, an inhibition of CRE DNA binding activity. Characterization of nuclear binding proteins further revealed that NF-AT and, to a lesser extent, NF-kappaB DNA binding activities were also suppressed. These studies demonstrate that F-Me-AEA modulates T cell function in a similar manner to plant-derived and endogenous cannabinoids and therefore can be utilized as an amidase- and hydrolysis-resistant endogenous cannabinoid.
Kaplan
Barbara L F
BL
Department of Pharmacology and Toxicology and Center for Integrative Toxicology, Michigan State University, East Lansing, MI 48824, USA.
Ouyang
Yanli
Y
Herring
Amy
A
Yea
Sung Su
SS
Razdan
Raj
R
Kaminski
Norbert E
NE
D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism* / Q000494:pharmacology*; D063386:Cannabinoid Receptor Agonists; D002186:Cannabinoids / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000037:antagonists & inhibitors; D004247:DNA / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D024202:Electrophoretic Mobility Shift Assay; D063388:Endocannabinoids; D005260:Female; D015870:Gene Expression; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors; D053284:Polyunsaturated Alkamides; D012333:RNA, Messenger / Q000096:biosynthesis; D043882:Receptors, Cannabinoid / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D014157:Transcription Factors / Q000037:antagonists & inhibitors
C443759:2-methylarachidonyl-2'-fluoroethylamide; D001095:Arachidonic Acids; D063386:Cannabinoid Receptor Agonists; D002186:Cannabinoids; D004268:DNA-Binding Proteins; D063388:Endocannabinoids; D007376:Interleukin-2; D016328:NF-kappa B; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D053284:Polyunsaturated Alkamides; D012333:RNA, Messenger; D043882:Receptors, Cannabinoid; D014157:Transcription Factors; D004247:DNA; D000242:Cyclic AMP; C078814:anandamide