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Lack of t(14;18) polymerase chain reaction-positive cells in highly
purified CD34+ cells and their CD19 subsets in patients with follicular
lymphoma
MT Voso, S Hohaus, M Moos and R Haas
Department of Internal Medicine V, University of Heidelberg and German
Cancer Research Center.
Follicular lymphoma (FL) is characterized in a significant proportion of
cases by the t(14;18) chromosomal translocation, which results in the
juxtaposition of the oncogene bcl-2 to the joining region of the
immunoglobulin heavy chain (IgH) gene. Molecular sequence analysis
indicates that the t(14;18) rearrangement occurs in a B-lymphoid progenitor
cell at the time of IgH rearrangement. We were interested whether
hematopoietic stem and progenitor cells as characterized by CD34 expression
bear the translocation. Bone marrow (BM)-CD34+ cells were enriched from 14
patients with FL whose BM was known to be positive for bcl-2/IgH (major
breakpoint region [MBR]). Six patients were in complete remission (CR), two
patients were in partial remission (PR), and six patients had active
disease. Six patients had histological BM involvement when the samples were
obtained. Using an immunomagnetic selection device (MINI-MACS), a mean
purity of 88.7% +/- 4% CD34+ cells was achieved. The CD34+ cells were
further enriched by fluorescence activated cell sorting (FACS) using CD34
fluorescein isothiocyanate (FITC)- and CD19 phycoerythrin (PE)-conjugated
antibodies. The IgH gene was rearranged in the CD34+/CD19+ cell subset of
all patients assessed by polymerase chain reaction (PCR). This population
is thought to represent the progenitor stage at which the bcl-2/IgH
translocation occurs. The unseparated BM mononuclear cell fraction from all
14 patients was positive for bcl-2/IgH using a nested PCR, but the BM-CD34+
cell fraction and the respective CD34+/CD19+ subset were negative in 13 of
these 14 patients. The one patient with a positive PCR signal in the CD34+
cell subset had a relapse with BM involvement. We conclude that CD34+
progenitor cells including CD34+/CD19+ B-cell progenitors are not involved
in the malignant cell clone. These data are in agreement with a transgenic
mouse model, which indicates that the malignant phenotype in FL is
sustained by mature B cells.
Volume 89,
Issue 10,
pp. 3763-3768,
05/15/1997
Copyright © 1997 by The American Society of Hematology
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