The number of memory B cells in peripheral blood has been assayed in various diseases by using CD27 as a memory B-cell marker. However, the defining differences of characteristic and function between the two memory B-cell subpopulations separated by immunoglobulin (Ig)D expression remain to be clearly elucidated. We analyzed here IgD(+)CD27(+) B cells (circulating B cells 2, cB2) and IgD(-)CD27(+) memory B cells (cB3) in comparison with IgD(+)CD27(-) naive B cells (cB1). cB2 were found to be morphologically similar to cB3 with abundant cytoplasm, whereas cB3 expressed CD80, CD86, and CD95 on their surface more predominantly than cB2. A majority of cB2 expressed both IgD and IgM, and cB3 expressed IgA or IgG. Mature gamma1 and gamma2 transcripts were found in cB3, but at very low levels in cB2, and activation-induced cytidine deaminase (AID) mRNA expression was recognized only in cB3. The frequencies of somatic hypermutation in cB2 and cB3 were comparable levels studied by VH5. cB2 did not shift to cB3 in vitro by the stimuli such as via B-cell receptor or CD40. cB2 produced large amounts of IgM predominantly and promptly, which is in accordance with the known characteristics of memory B cells. Taken together, although cB2 are unclass-switched, cB2 have the functions of memory B cells and are not in the process of transition from naive to switched memory B cells, playing a crucial role in secondary immune response by producing high-affinity IgM in the early phase of infections.