Extracellular ATP triggers IL-1 beta release by activating the purinergic P2Z receptor of human macrophages.
Extracellular ATP (ATPe) is known to cause release of processed IL-1 beta from LPS-treated macrophages and microglial cells. IL-1 beta release is fast and thought to be associated with cell death. We have reinvestigated this process to identify 1) the purinergic receptor involved; 2) the relationship to cell death; and 3) pharmacologic agonists or antagonists able to modulate IL-1 beta release. Our data confirm that ATPe is a powerful stimulus for IL-1 beta release from LPS-treated human macrophages; however, we also show that IL-1 beta release is not necessarily associated with cell death, as it occurs at lower ATP concentrations and much earlier than leakage of cytoplasmic markers. The selective purinergic P2Z receptor agonist benzoylbenzoyl ATP was at least one order of magnitude more powerful than ATP, but also had a strong cytotoxic effect. 2-Methylthio-ATP was equipotent as ATPe at the optimal concentration of 1 mM, but markedly inhibitory at higher concentrations. The irreversible P2Z blocker-oxidized ATP completely inhibited ATPe-induced IL-1 beta release. IL-1 beta release also was inhibited by increasing the K+ concentration of the incubation medium. These data suggest that ATPe triggers IL-1 beta via the purinergic P2Z receptor recently shown to be expressed by human macrophages and identified as a new member of the P2X family (P2X7), and provide pharmacologic tools for the modulation of IL-1 beta release in vitro and, possibly, in vivo.
Ferrari
D
D
Institute of General Pathology, University of Ferrara, Italy.
Chiozzi
P
P
Falzoni
S
S
Dal Susino
M
M
Melchiorri
L
L
Baricordi
O R
OR
Di Virgilio
F
F
D000255:Adenosine Triphosphate / Q000502:physiology*; D015415:Biomarkers; D016923:Cell Death / Q000187:drug effects / Q000276:immunology; D003593:Cytoplasm / Q000201:enzymology; D004306:Dose-Response Relationship, Immunologic; D010450:Endopeptidases / Q000378:metabolism; D005110:Extracellular Space / Q000502:physiology*; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism*; D008264:Macrophages / Q000201:enzymology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D018048:Receptors, Purinergic P2 / Q000187:drug effects / Q000378:metabolism*; D058486:Receptors, Purinergic P2X7; D053148:Caspase 3
D000255:Adenosine Triphosphate; D015415:Biomarkers; D010450:Endopeptidases; D007375:Interleukin-1; D009419:Nerve Tissue Proteins; D018048:Receptors, Purinergic P2; D058486:Receptors, Purinergic P2X7; C546468:P2RX7 protein, human; C505410:Casp3 protein, rat; D053148:Caspase 3