Adenosine Receptor Agonists in Lymphocytes

In lymphocytes, the potency of the well-known A_2A and A₃ adenosine agonists CGS 21680 and Cl-IB-MECA (0.1 nM–1 μM) was investigated. Cells were seeded in 96-well white half-area microplate (Perkin Elmer, Boston, MA, USA) in a stimulation buffer composed of Hank Balanced Salt Solution, 5 mM HEPES, 0.5 mM Ro 20-1724, 0.1% BSA. Forskolin (1 μM) was used to stimulate adenylyl cyclase activity after the addition of Cl-IB-MECA. cAMP levels were then quantified by using the AlphaScreen cAMP Detection Kit (Perkin Elmer) following the manufacturer’s instructions. At the end of the experiments, plates were read with the Perkin Elmer EnSight Multimode Plate Reader.