Abstract
Pseudomonas aeruginosa, a gram-negative multidrug-resistant (MDR) opportunist, belongs to the ESKAPE group of pathogens associated with the highest risk of mortality. Neutrophil swarming is a host defense strategy triggered by larger threats, where neutrophil swarms contain and clear damage/infection. Current ex vivo models designed to study neutrophil-pathogen interactions largely focus on individual neutrophil engagement with bacteria and fail to capture neutrophil swarming. Here, we report an ex vivo model that reproducibly elicits neutrophil swarming in response to bacterial aggregates. A rapid and robust swarming response follows engagement with pathogenic targets. Components of the type III secretion system (T3SS), a critical P. aeruginosa virulence determinant, are involved in swarm interaction. This ex vivo approach for studying neutrophil swarming in response to large pathogen targets constitutes a valuable tool for elucidating host-pathogen interaction mechanisms and for evaluating novel therapeutics to combat MDR infections.