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Monitoring of tumor cell purging after highly efficient immunomagnetic
selection of CD34 cells from leukapheresis products in breast cancer
patients: comparison of immunocytochemical tumor cell staining and reverse
transcriptase-polymerase chain reaction
MY Mapara, IJ Korner, M Hildebrandt, R Bargou, D Krahl, P Reichardt and B Dorken
Department of Internal Medicine (Medical Oncology and Tumorimmunology),
Humboldt University, Virchow Klinikum-Robert Rossle Klinik, Berlin,
Germany.
We studied the efficiency of indirect tumor cell purging via enrichment of
CD34+ hematopoietic progenitor cells from leukapheresis products (LP) in
breast cancer patients based on immunomagnetic selection of CD34+ cells.
Detection of tumor cells was made by immunocytochemical staining. In
addition, we evaluated the capacity of cytokeratin 19 (CK19)- and a novel
epidermal growth factor receptor (EGF-R)-specific reverse
transcriptase-polymerase chain reaction (RT-PCR) for monitoring tumor cell
depletion. LP from 13 breast cancer patients were analyzed. Twenty-three
CD34 selection procedures were performed. A median of 1.4 x 10(10) total
nucleated cells ([TNC] range, 0.88 to 3.5 x 10(10)) with a median CD34
purity of 2.5% (range, 0.4% to 6.3%) were entered into the selection
procedure. Immunomagnetic CD34 enrichment resulted in a median purity of
83.3% (range, 45% to 95.4%) and a median recovery of 73.2% (range, 22% to
95%). Retransfusion of CD34-selected cells after high-dose chemotherapy
resulted in a rapid and sustained hematologic recovery, reaching an
absolute neutrophil count of 500/microL at day +10 and platelet count of
20,000/microL at day +11. Tumor cell depletion was quantified by
immunocytochemical detection of CK19- positive cells. By this method, a
median tumor cell depletion of 1.9 log (range, 0.7 to > 3 log) could be
demonstrated. Immunocytochemical detection of tumor cells was more
sensitive than RT-PCR, yielding positive results in 81% of LP (17 to 21)
versus 58% positive LP (10 of 17). However, EGF-R-based RT-PCR was much
more sensitive than CK19- based RT-PCR (10 of 17 v 1 of 17). Despite highly
efficient CD34 selection, tumor cells were still detectable after CD34
enrichment using immunocytochemistry and EGF-R-specific RT-PCR. Thus, this
novel EGF-R-specific RT-PCR appears to be of value as an additional method
to detect contaminating breast cancer cells within LP.
Volume 89,
Issue 1,
pp. 337-344,
01/01/1997
Copyright © 1997 by The American Society of Hematology
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