title: Mechanism of cytokine inhibition of beta-adrenergic agonist stimulation of cyclic AMP in rat cardiac myocytes. Impairment of signal transduction.
abstract: Studies conducted in our laboratory have demonstrated that activated immune cells produce a soluble inhibitor(s) of cardiac myocyte contractile and cyclic AMP (cAMP) responses to beta-adrenergic stimulation. To examine the mechanism of this effect, metabolic assays were conducted on cultured rat cardiac myocytes incubated in the presence and absence of supernatants harvested from rat activated splenocyte cultures. Intracellular cAMP accumulation in response to isoproterenol was inhibited by up to 74% in a dose-dependent fashion by conditioned media containing soluble cytokines from activated immune cells. By use of myocyte cultures in which contaminating nonmyocyte proliferation was inhibited by nonlethal irradiation, this phenomenon was shown to be independent of mitogenic effects. Isobutylmethylxanthine, a phosphodiesterase inhibitor, did not ablate cytokine-induced inhibition of cAMP accumulation. Parameters of beta-adrenergic receptor binding and affinity were also unaffected. cAMP suppression was maintained after cholera toxin stimulation of cAMP production via stimulatory G protein ADP-ribosylation. cAMP inhibition was not apparent when cells were stimulated with forskolin, a direct adenylate cyclase activator. Importantly, pertussis toxin treatment significantly ablated cytokine-induced cAMP inhibition. Thus, interference with agonist-occupied beta-adrenergic receptor coupling to adenylate cyclase to produce cAMP and subsequent contractile responses is induced by a factor(s) elaborated by activated immune cells. This interference occurs at the level of signal transduction across the membrane, can be overridden by pertussis toxin, and may involve changes in the coupling of the stimulatory/inhibitory G proteins to adenylate cyclase. These results demonstrate a novel mechanism of cytokine-induced myocyte dysfunction and may have important pathophysiological ramifications in immune-mediated myocardial diseases.
authors.affiliation: Department of Medicine, Jewish Hospital of St. Louis, Washington University Medical Center, MO 63110.
authors.forename: M K
authors.initials: MK
authors.lastname: Chung
authors.forename: T S
authors.initials: TS
authors.lastname: Gulick
authors.forename: R E
authors.initials: RE
authors.lastname: Rotondo
authors.forename: G F
authors.initials: GF
authors.lastname: Schreiner
authors.forename: L G
authors.initials: LG
authors.lastname: Lange
mesh_terms: D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000032:analysis; D000818:Animals; D001685:Biological Factors / Q000494:pharmacology*; D002478:Cells, Cultured; D003470:Culture Media; D000242:Cyclic AMP / Q000032:analysis*; D016207:Cytokines; D007545:Isoproterenol / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D009206:Myocardium / Q000032:analysis / Q000166:cytology / Q000378:metabolism*; D037342:Pertussis Toxin; D011869:Radioligand Assay; D051381:Rats; D011943:Receptors, Adrenergic, beta / Q000187:drug effects; D015398:Signal Transduction; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
chemical_list: D037361:Adenylate Cyclase Toxin; D001685:Biological Factors; D003470:Culture Media; D016207:Cytokines; D011943:Receptors, Adrenergic, beta; D010566:Virulence Factors, Bordetella; D000242:Cyclic AMP; D037342:Pertussis Toxin; D000262:Adenylyl Cyclases; D007545:Isoproterenol