Preparation and Inflammasome Activation of Murine Bone Marrow–Derived and Resident Peritoneal Macrophages

Cholera toxin (CT), secreted by Vibrio cholerae, not only causes cholera but also functions as an immune adjuvant. Macrophages, which respond to a variety of immune adjuvants, are quite heterogenous and their development and function depend on the tissues where they are localized. We have characterized the effects of the B subunit of CT (CTB) on two types of murine macrophages, that is, bone marrow–derived macrophages (BMMs) and resident peritoneal macrophages (rPMs). CTB could induce production of interleukin-1β (IL-1β) from both macrophages in synergy with lipopolysaccharides. However, underlying molecular mechanisms for IL-1β induction were different. Here, we describe the protocols for preparation and stimulation of BMMs and rPMs.