B-1 cells are innate-like lymphocytes that generate natural, polyreactive antibodies with important functions in tissue homeostasis and immune defense. While B-1-cell frequencies in secondary lymphoid tissues are low, relative high frequencies are found within peritoneal and pleural cavities of mice, including both CD5(+) B-1a and CD5(-) B-1b cells. They represent reservoirs of B-1 cells that can be activated for migration to lymphoid tissues to secrete antibodies and/or cytokines. Here, we outline efficient methods for the extraction and magnetic isolation of B-1a cells from the peritoneal and pleural cavities and the separation and phenotypic characterization of B-1a and B1-b cells by flow cytometry.
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