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

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Abstract
Using a method which allows simultaneous flow cytometric detection of cell surface markers and 5-bromo-2'-deoxyuridine (BrdU) incorporation, the distribution and proliferative behavior of B lineage subpopulations was studied in intact adult mice. In the bone marrow we could define two subsets of B cells on the basis of differential expression of the pan-B cell marker B220 and of membrane-associated μ and δ immunoglobulin heavy chains. B220dullμ+δ− B cells were found to emerge from rapidly dividing cells and probably represent B cells recently generated from B220dullμ− pie-B cells. In contrast, oniy few, If any, of the B220brightμ+δ+ B cells were labeled with BrdU after a period of 8 days, suggesting that these cells represent long-lived B cells residing in the bone marrow. Analysis of BrdU-Incorporation into splenic B cells showed that only 20% of these cells had gone through cell division during the preceding 8 days. Almost none of the B cells in the peritoneum, a large fraction of which belongs to the Ly1 B subset, were labeled with BrdU over a period of 7 days in 8-month-old animals.

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