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CD34+ cells in the blood of patients with multiple myeloma express CD19 and
IgH mRNA and have patient-specific IgH VDJ gene rearrangements
AJ Szczepek, PL Bergsagel, L Axelsson, CB Brown, AR Belch and LM Pilarski
Department of Oncology, University of Alberta, Edmonton, Canada.
Peripheral blood mononuclear cells (PBMC) from patients with multiple
myeloma (MM) are here shown to include 23% +/- 2% of CD34+ cells, the
majority of which coexpress CD19, as identified by a panel of 17 anti- CD34
antibodies. The expression of CD34 mRNA by sorted CD34+ PBMC from MM was
confirmed by in situ reverse transcriptase-polymerase chain reaction
(RT-PCR) with CD34-specific primers. The majority of CD34+ MM PBMC were
CD19+ cells that expressed mRNA for CD19 and for rearranged IgH as
identified with consensus IgH VDJ primers, as well as having cytoplasmic
Ig, definitively identifying them as B cells, in absolute numbers of 0.06
to 0.69 x 10(9)/L of blood. CD34 is largely absent from normal B cells. To
determine the clonal relationship of CD34+ B cells to autologous MM plasma
cells, IgH VDJ DNA rearrangements of sorted CD34+ MM blood B cells were
amplified by nested PCR using consensus primers followed by Southern
blotting with allele-specific oligonucleotides for 7 MM patients, and
clonotypic IgH mRNA expression was assessed for 4 MM patients using
quantitative patient-specific in situ RT-PCR. For 9 of 11 myeloma patients
tested, CD34+ blood B cells included IgH gene rearrangements or expressed
IgH mRNA identical to that of autologous bone marrow plasma cells. For 4 of
4 MM patients, 74% to 94% of individual sorted CD34+19+ B cells expressed
clonotypic IgH mRNA, as detected by in situ RT-PCR with patient-specific
primers. Clonotypic IgH VDJ sequences were absent from B cells of unrelated
MM patients and of normal donors. Clonotypic CD34+ B cells were detected
before, during, and after treatment, and during relapse. Our results
indicate a clonal relationship between CD34+ MM B cells and malignant
plasma cells. We speculate that CD34 may play an important role in the
biology of myeloma by facilitating extravasation from blood and thus spread
of myeloma through the skeletal system.
Volume 89,
Issue 5,
pp. 1824-1833,
03/01/1997
Copyright © 1997 by The American Society of Hematology
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