Memory B lymphocytes in human tonsil do not express surface IgD

To clarify the phenotype of memory B lymphocytes, we have determined the frequency of somatic hypermutations in purified tonsil B cell populations. Our particular interest was the controversial question of whether any memory B-cells express IgD. Ig heavy chain gene rearrangements that used the nonpolymorphic VH6 gene were amplified by PCR, cloned, and sequenced. All eight sequences obtained from the surface IgD (sIgD)-negative fraction contained point mutations, with frequency of one mutation in every 24 bases of sequence. In contrast, only 4 of 12 sequences obtained from the sIgD-positive fraction contained point mutations, with a mutation frequency of one in 600. This frequency was similar to that found for cord blood B cells (one in 550), a population that does not contain memory B cells. These results indicate that although memory B cells are present in the sIgD-negative fraction, no memory cells can be detected in the sIgD-positive fraction of tonsil B lymphocytes.