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*
Servizio di Immunologia Clinica and
Servizio di Citometria Centro Biotechnologie Avanzate, Istituto Nazionale per la Ricerca sul Cancro, IST, Genova, Italy;
Dipartimento di Oncologia, Biologia e Genetica, Università degli Studi di Genova, Genova, Italy;
§
Divisione ORL, Istituto Giannina Gaslini, Genova, Italy; and
¶
Department of Medicine, North Shore University Hospital and New York University School of Medicine, Manhasset, NY, 11030
The VH4 genes expressed by both resting and in
vivo-activated subepithelial (SE) B cells from human tonsils were
studied. Resting SE B cells were subdivided according to the presence
(IgDlow) or absence (IgM-only) of surface IgD. CD27 was
abundant on activated SE B cells and low on resting IgM-only B cells.
Resting IgDlow SE B cells could be subdivided into
CD27low and CD27high cell fractions. Resting
IgDlow SE B cells displayed VH4 genes with a
substantial number of mutations (13/29 of the molecular clones were
mutated), whereas 25/26 of the clones from resting IgM-only SE B cells
were unmutated. Moreover, mutated VH4 genes were detected
mainly within the CD27high cell fraction of the
IgDlow SE B cells. Several identical unmutated
VH4DJH sequences (11/32) were found in
different molecular clones from resting IgM-only SE B cells, suggesting
local cellular expansion. Both unmutated (14/25) and mutated (11/25)
sequences were found in µ transcripts of activated SE B cells.
Extensive mutation was observed in the 
transcripts of activated SE
B cells. Therefore, SE B cells are heterogeneous, being comprised of B
cells with mutated Ig VH4 genes, that are Ag-experienced B
cells, and a subset of B cells with unmutated VH4 genes
that are either virgin cells or cells driven by Ags that did not induce
or select for V gene mutations.
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