Cannabinoid inhibition of adenylate cyclase-mediated signal transduction and interleukin 2 (IL-2) expression in the murine T-cell line, EL4.IL-2

J Biol Chem. 1996 May 31;271(22):13175-83. doi: 10.1074/jbc.271.22.13175.


Cannabinoid receptors negatively regulate adenylate cyclase through a pertussis toxin-sensitive GTP-binding protein. In the present studies, signaling via the adenylate cyclase/cAMP pathway was investigated in the murine thymoma-derived T-cell line, EL4.IL-2. Northern analysis of EL4.IL-2 cells identified the presence of 4-kilobase CB2 but not CB1 receptor-subtype mRNA transcripts. Southern analysis of genomic DNA digests for the CB2 receptor demonstrated identical banding patterns for EL4.IL-2 cells and mouse-derived DNA, both of which were dissimilar to DNA isolated from rat. Treatment of EL4.IL-2 cells with either cannabinol or Delta9-THC disrupted the adenylate cyclase signaling cascade by inhibiting forskolin-stimulated cAMP accumulation which consequently led to a decrease in protein kinase A activity and the binding of transcription factors to a CRE consensus sequence. Likewise, an inhibition of phorbol 12-myristate 13-acetate (PMA)/ionomycin-induced interleukin 2 (IL-2) protein secretion, which correlated to decreased IL-2 gene transcription, was induced by both cannabinol and Delta9-THC. Further, cannabinoid treatment also decreased PMA/ionomycin-induced nuclear factor binding to the AP-1 proximal site of the IL-2 promoter. Conversely, forskolin enhanced PMA/ionomycin-induced AP-1 binding. These findings suggest that inhibition of signal transduction via the adenylate cyclase/cAMP pathway induces T-cell dysfunction which leads to a diminution in IL-2 gene transcription.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenylyl Cyclase Inhibitors*
  • Adenylyl Cyclases / metabolism*
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cannabinol / pharmacology*
  • Cell Line
  • Cyclic AMP / metabolism
  • DNA Primers
  • Dronabinol / pharmacology*
  • Female
  • Interleukin-2 / genetics*
  • Ionomycin / pharmacology
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • RNA, Messenger / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction*
  • T-Lymphocytes / metabolism
  • Tetradecanoylphorbol Acetate / pharmacology
  • Trans-Activators / metabolism
  • Transcription Factor AP-1 / metabolism


  • Adenylyl Cyclase Inhibitors
  • DNA Primers
  • Interleukin-2
  • RNA, Messenger
  • Trans-Activators
  • Transcription Factor AP-1
  • Ionomycin
  • Dronabinol
  • Cannabinol
  • Cyclic AMP
  • Adenylyl Cyclases
  • Tetradecanoylphorbol Acetate