Protocol for isolating and characterizing effector functions of murine bone marrow-derived eosinophils following bacterial challenge

Understanding eosinophil-associated mechanisms, especially in the context of infection, has been steadily increasing. Here, we present a protocol for isolating and differentiating murine bone marrow-derived eosinophils (bmEos) followed by multi-well plate assays to evaluate eosinophilic responses to bacterial challenge. We describe steps for evaluating eosinophil effector functions including cytotoxicity, bactericidal activity, and cytokine production through multiplex ELISA panels. We use a Bordetella bronchiseptica infection model, but this approach can be modified for an array of microorganisms and different experimental settings. For complete details on the use and execution of this protocol, please refer to First et al.¹.