title: Rolipram, salbutamol and prostaglandin E2 suppress TNFalpha release from human monocytes by activating Type II cAMP-dependent protein kinase.
abstract: The extent to which cAMP-dependent protein kinase (PKA) mediates the inhibitory effects of cAMP-elevating drugs on tumour necrosis factor (TNF) alpha release from lipopolysaccharide (LPS)-stimulated human monocytes is equivocal. Here, we have investigated the role of this kinase by exploiting the ability of certain novel cAMP analogues to inhibit or activate PKA and the recently described cAMP-guanine nucleotide-exchange factors (GEFs). Pre-treatment of monocytes with Rp-8-Br-cAMPS, a selective inhibitor of Type I PKA that has no effect on basal or stimulated Rap1 (a downstream effector of cAMP-GEFs) activity, potentiated LPS-induced TNFalpha output but had little or no effect on the suppression of this cytokine effected by rolipram (a PDE4 inhibitor), prostaglandin (PG) E2 and salbutamol (a beta2-adrenoceptor agonist). In contrast, Rp-8-pCPT-cAMPS, which selectively blocks Type II PKA with only weak activity against Rap1, significantly antagonised or abolished the inhibitory effect of these cAMP-elevating agents. Pre-treatment of monocytes with 8-pCPT-2'-O-Me-cAMPS, a potent activator of cAMP-GEFs, failed to suppress TNFalpha output at concentrations known to profoundly activate Rap1. Collectively, these results indicate that cAMP-elevating drugs suppress TNFalpha release from LPS-stimulated human monocytes by activating PKA independently of cAMP-GEFs. Furthermore, by using phosphorothioate cAMP analogue PKA inhibitors we provide evidence that the Type II PKA isoenzyme is functionally the most important.
authors.affiliation: Dermatology Section, Division of Medicine, Faculty of Medicine, Hammersmith Hospital, Imperial College London, DuCane Road, London W12 ONN, UK.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Seldon
authors.forename: Koremu K
authors.initials: KK
authors.lastname: Meja
authors.forename: Mark A
authors.initials: MA
authors.lastname: Giembycz
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000420:Albuterol / Q000494:pharmacology*; D015153:Blotting, Western; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015232:Dinoprostone / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000378:metabolism; D020889:Rolipram / Q000494:pharmacology*; D013873:Thionucleotides / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: C473217:8-(4-chloro-phenylthio)-2'-O-methyladenosine-3'-5'-cyclic monophosphate; C105593:8-bromoadenosine-3',5'-cyclic monophosphorothioate; D008070:Lipopolysaccharides; D021122:Protein Subunits; D013873:Thionucleotides; D014409:Tumor Necrosis Factor-alpha; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000242:Cyclic AMP; D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases; D020889:Rolipram; D015232:Dinoprostone; D000420:Albuterol