Purification and Immune Phenotyping of B-1 Cells from Body Cavities of Mice.

B-1 cells are fetal-origin B lymphocytes with unique developmental and functional characteristics that can generate natural, polyreactive antibodies with important functions in tissue homeostasis and immune defense. While B-1 cell frequencies in bone marrow and secondary lymphoid tissues are low, relative high frequencies exist within peritoneal and pleural cavities of mice, including both CD5 ⁺ and CD5 ^- B-1 cells. These cells represent B-1 reservoirs that, when activated, migrate to lymphoid tissues to secrete antibodies and/or cytokines. Here, we outline efficient methods for the extraction and magnetic isolation of CD5 ⁺ B-1 cells from the peritoneal and pleural cavities as well as the separation and phenotypic characterization of CD5 ⁺ and CD5 ^- B-1 cells by flow cytometry.