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Department of Immunology, Duke University Medical Center, Durham, NC 27710
We have recently demonstrated that a novel somatically mutated B220- memory B cell subset rapidly dominates the secondary immune response to (4-hydroxy-3-nitrophenyl) acetyl (NP). Upon adoptive transfer with Ag, B220+NP+ memory B cells produce large numbers of B220-NP+ B cells that can rapidly differentiate into plasma cells. Therefore, it is not clear whether the novel B220- memory compartment is a consequence of secondary Ag challenge or whether it develops as a stable memory subset after initial Ag challenge. In this study, we demonstrate the gradual emergence of B220-NP+ B cells in the spleen to maximal numbers 3 wk after initial Ag exposure. Like their B220+ counterparts, the B220- B cells initially appear unmutated at days 5–7; however, the majority rapidly accumulate affinity increasing mutations by days 9–14 of the primary immune response. More extensive cell surface phenotype (GL7-BLA-1-CD24-CD43+) argues strongly against germinal center localization and direct analysis in situ places a cohort of B220-CD11b+NP+ B cells in the red pulp of the spleen and not in the MZs. These data provide direct evidence for the development of B220- memory B cells as a unique cellular consequence of primary Ag exposure. The cellular dynamics and molecular attributes of these unique memory B cells suggest they are distinct cellular products of the germinal center reaction in the primary response and are maintained long-term in the spleen and bone marrow.
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