Flow cytometric analysis of cell proliferation dynamics in the B cell compartment of the mouse

Using a method which allows simultaneous flow cytometric detectionof cell surface markers and 5-bromo-2'-deoxyuridine (BrdU) incorporation,the distribution and proliferative behavior of B lineage subpopulationswas studied in intact adult mice. In the bone marrow we coulddefine two subsets of B cells on the basis of differential expressionof the pan-B cell marker B220 and of membrane-associated µand immunoglobulin heavy chains. B220^dullµ^+– Bcells were found to emerge from rapidly dividing cells and probablyrepresent B cells recently generated from B220^dullµ^–pie-B cells. In contrast, oniy few, If any, of the B220^brightµ⁺⁺B cells were labeled with BrdU after a period of 8 days, suggestingthat these cells represent long-lived B cells residing in thebone marrow. Analysis of BrdU-Incorporation into splenic B cellsshowed that only 20% of these cells had gone through cell divisionduring the preceding 8 days. Almost none of the B cells in theperitoneum, a large fraction of which belongs to the Ly1 B subset,were labeled with BrdU over a period of 7 days in 8-month-oldanimals.