Signaling dynamics distinguish high- and low-priority neutrophil chemoattractant receptors

Human neutrophils respond to multiple chemoattractants to guide their migration from the vasculature to sites of infection and injury, where they clear pathogens and amplify inflammation. To properly focus their responses during this complex navigation, neutrophils prioritize pathogen- and injury-derived signals over long-range inflammatory signals, such as the leukotriene LTB4, secreted by host cells. Different chemoattractants can also drive qualitatively different modes of migration even though their receptors couple to the same Gα i family of G proteins. Here, we used live-cell imaging to demonstrate that the responses differed in their signaling dynamics. Low-priority chemoattractants caused transient responses, whereas responses to high-priority chemoattractants were sustained. We observed this difference in both primary neutrophils and differentiated HL-60 cells, for downstream signaling mediated by Ca 2+ , a major regulator of secretion, and Cdc42, a primary regulator of polarity and cell steering. The rapid attenuation of Cdc42 activation in response to LTB4 depended on the phosphorylation sites Thr 308 and Ser 310 in the carboxyl-terminal tail of its receptor LTB4R in a manner independent of endocytosis. Mutation of these residues to alanine impaired chemoattractant prioritization, although it did not affect chemoattractant-dependent differences in migration persistence. Our results indicate that distinct temporal regulation of shared signaling pathways distinguishes between receptors and contributes to chemoattractant prioritization. Neutrophils prioritize chemoattractant inputs according to the duration of the signaling output. Neutrophils are simultaneously exposed to multiple chemoattractants that bind to receptors that activate Gα i proteins. Lundgren et al . investigated how neutrophils prioritize responses to one chemoattractant over another. In primary human neutrophils and a neutrophil cell line, the signaling initiated by high-priority chemoattractants (such as those released from sites of infection or damage) lasted longer than that induced by low-priority chemoattractants (such as those released from sites of inflammation). The rapid attenuation of signaling downstream of the receptor for a low-priority chemoattractant required the phosphorylation of two sites in the C-terminal tail of the receptor. Together, these results reveal how neutrophils rank chemoattractants that activate a common signaling pathway. —Wei Wong