Monitoring of tumor cell purging after highly efficient immunomagnetic selection of CD34 cells from leukapheresis products in breast cancer patients: comparision of immunocytochemical tumor cells staining and RT-PCR

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Item Type:	Article
Title:	Monitoring of tumor cell purging after highly efficient immunomagnetic selection of CD34 cells from leukapheresis products in breast cancer patients: comparision of immunocytochemical tumor cells staining and RT-PCR
Creators Name:	Mapara, M.Y., Koerner, I.J., Hildebrandt, M., Bargou, R.C., Krahl, D., Reichardt, P. and Doerken, B.
Abstract:	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.
Keywords:	CD34 Antigens, Antineoplastic Combined Chemotherapy Protocols, Breast Neoplasms, Cisplatin, Complementary DNA, Neoplasm DNA, Epirubicin, Etoposide, Evaluation Studies as Topic, Granulocyte Colony-Stimulating Factor, Hematopoietic Stem Cell Transplantation, Ifosfamide, Immunoenzyme Techniques, Immunomagnetic Separation, Keratins, Leukapheresis, Melphalan, Neoplasm Proteins, Circulating Neoplastic Cells, Polymerase Chain Reaction, Messenger RNA, Neoplasm RNA, Epidermal Growth Factor Receptor, Transplantation Conditioning, Biological Tumor Markers
Source:	Blood
ISSN:	0006-4971
Publisher:	American Society of Hematology
Volume:	89
Number:	1
Page Range:	337-344
Date:	1 January 1997
Official Publication:	http://bloodjournal.hematologylibrary.org/cgi/content/abstract/89/1/337
PubMed:	View item in PubMed

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