Although extracellular ATP is abundant at sites of inflammation, its role in activating inflammasome signalling in neutrophils is not well characterized. In the current study, we demonstrate that human and murine neutrophils express functional cell-surface P2X₇R, which leads to ATP-induced loss of intracellular K⁺, NLRP3 inflammasome activation and IL-1β secretion. ATP-induced P2X₇R activation caused a sustained increase in intracellular [Ca²⁺], which is indicative of P2X₇R channel opening. Although there are multiple polymorphic variants of P2X₇R, we found that neutrophils from multiple donors express P2X₇R, but with differential efficacies in ATP-induced increase in cytosolic [Ca²⁺]. Neutrophils were also the predominant P2X₇R-expressing cells during Streptococcus pneumoniae corneal infection, and P2X₇R was required for bacterial clearance. Given the ubiquitous presence of neutrophils and extracellular ATP in multiple inflammatory conditions, ATP-induced P2X₇R activation and IL-1β secretion by neutrophils likely has a significant, wide ranging clinical impact.