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.