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P2X₇ Nucleotide Receptor Mediation of Membrane Pore Formation and Superoxide Generation in Human Promyelocytes and Neutrophils¹

Byung-Chang Suh^*, Jong-So Kim^*, Uk Namgung^*, Hyunjung Ha and Kyong-Tai Kim^2,*

^* Department of Life Science, Division of Molecular and Life Science, Pohang University of Science and Technology, Hyoja-Dong, Pohang, Republic of Korea; and Division of Life Sciences, Chungbuk National University, Cheongju, Republic of Korea

The P2X₇ receptor, which induces cation channel opening imparting significant permeability to Ca²⁺ and pore formation with changes in the plasma membrane potential, has been known to be rather restrictedly expressed in cells of the macrophage lineage including dendrites, mature macrophages, and microglial cells. However, we show here that the P2X₇ receptor is also expressed in cells of granulocytic lineage such as HL-60 promyelocytes, granulocytic differentiated cells, and neutrophils. Exposure of these cells to 2',3'-O-(4-benzoyl)benzoyl-ATP (BzATP) triggered intracellular Ca²⁺ rise through the mediation of phospholipase C-independent and suramin-sensitive pathways. BzATP also induced depolarization of the plasma membrane in the absence of extracellular Ca²⁺, whereas it hyperpolarized the cells in the presence of external Ca²⁺, probably in part through the activation of Ca²⁺-activated K⁺ channels. However, the hyperpolarization phenomenon was markedly attenuated in differentiated HL-60 cells and neutrophils. RT-PCR and Northern blot analysis revealed the presence of P2X₇ receptors on both HL-60 and neutrophil-like cells. This was further confirmed by pore formation through which the uptake of Lucifer yellow and YO-PRO1 occurred on BzATP treatment. BzATP stimulated in a concentration-dependent manner the production of superoxide in differentiated HL-60 cells via a pathway partially dependent on extracellular Ca²⁺. Moreover, in human neutrophils, BzATP was a more effective inducer of superoxide generation than PMA. Taken together, this is a first demonstration of the expression of P2X₇ receptors on neutrophils, which shows that the receptor is functionally involved in the defense mechanism by activation of the respiratory burst pathway.

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