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Blood, Vol. 96 No. 3 (August 1), 2000:
pp. 996-1005
Pharmacologic properties of P2Z/P2X7
receptor characterized in murine dendritic cells: role on
the induction of apoptosis
Oscar Kenji Nihei,
Antonio Carlos Campos de Carvalho,
Wilson Savino, and
Luiz Anastacio Alves
From the Laboratory on Thymus Research, Department of Immunology,
Oswaldo Cruz Institute, FIOCRUZ; and the Laboratory of Excitable
Membranes, "Carlos Chagas Filho" Institute of Biophysics, Federal
University of Rio de Janeiro, Rio de Janeiro, Brazil.
In the immune system, extracellular adenosine 5'-triphosphate
(ATP) mediates a variety of effects mainly through activation of a
particular receptor subtype, the pore-forming
P2Z/P2X7 purinoceptor. This purinergic receptor
has been described chiefly in cells of hemopoietic origin such as T
cells, thymocytes, monocytes, macrophages, and phagocytic cells of
thymic reticulum. In this study, we characterized the
P2Z/P2X7 purinoceptor and the ATP-mediated
apoptosis in murine spleen-derived dendritic cells (DCs). Dye uptake
and apoptosis were evaluated by flow cytometry. ATP-treated DCs were
permeable to different low-molecular-weight fluorescent probes such as
ethidium bromide, YO-PRO 1, and lucifer yellow. Such an effect was
dose-dependent (EC50: 721 µmol/L); mediated by the fully
anionic agonist (ATP4 ); and specifically stimulated by
ATP, BzATP, and ATP S. Additionally, an ATP-induced increase in
intracellular calcium was detected by microfluorometry. Furthermore,
ATP treatment induced a significant increase in apoptotic DCs
(64.46% ± 3.8%) when compared with untreated control cells
(34% ± 5.8%), as ascertained by the TdT-mediated dUTP nick end
labeling technique. Both ATP-induced DC permeabilization and apoptosis
were inhibited by oxidized ATP, a
P2Z/P2X7-specific antagonist. In conclusion, we
characterized the expression of the P2Z/P2X7
purinoceptor in murine spleen-derived DCs and described its role on
the induction of apoptosis.
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