Within Germinal Centers, Isotype Switching of Immunoglobulin Genes Occurs after the Onset of Somatic Mutation

Human tonsillar B cells were separated into naive IgD +CD38 −CD23 − (Bm1) and IgD +CD38 −CD23 + (Bm2), germinal center IgD −CD38 +CD23 − centroblasts (Bm3) and IgD −CD38 +CD77 − centrocytes (Bm4), and memory IgD −CD38 − (Bm5) subsets. Previous IgV H sequence analysis concluded that the triggering of somatic mutations occurs during the transition from Bm2 subset into the Bm3 subset. To determine the initiation of isotype switching, sterile transcript expression was analyzed by amplification, cloning, and sequencing. A selective sterile Iγ, Iα, and Iε expression was observed at centrocyte (Bm4) stage, suggesting that isotype switch is triggered within germinal centers, after somatic mutation is initiated within centroblasts (Bm3). Finally, the high level of 5′Sγ–Sμ3′ DNA switching circles observed in germinal center B cells indicates that within human tonsils, germinal center is a major location for isotype switching.