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Immunomagnetic tumor cell selection--implications for the detection of disseminated cancer cells
W Krüger1, C Datta, A Badbaran
1Bone Marrow Transplantation Center, Department of Transfusion Medicine, University Hospital Eppendorf, Hamburg, Germany. krueger@uke.uni-hamburg.de
Background:
The optimal method for the detection of disseminated epithelial cancer cells has not yet been found. The standard method, using immunocytochemistry, offers a sensitivity of up to 10(-6). Molecular methods such as cytokeratin-19 RT-PCR are about 10 times as sensitive, but they are hampered by interference such as illegitimate gene expression.
Study Design And Methods:
Immunomagnetic bead selection of epithelial cancer cells using conjugates directed against the human epithelial antigen (HEA) followed by immunocytochemistry testing was investigated in this trial.
Results:
No cytokeratin-positive cells could be enriched from 56 control samples. In 104 clinical samples of bone marrow aspirations, PBPC collections, and venous blood obtained from breast cancer patients, the cytokeratin-positive rate increased significantly, from 29.9 percent before selection to 54.8 percent after enrichment. Even the yield of detected cancer cells was significantly higher after selection. Up to 2.5 x 10(8) MNCs were easily processed. However, the mean cancer cell recovery after HEA enrichment was only 24.4 percent. Subsequently, selected epithelial cells were successfully immunophenotyped by use of a double-stain technique detecting cytokeratin-positive cells and the urokinase-like plasminogen activator receptor.
Conclusion:
HEA bead selection in combination with the standard immunocytochemistry method is a powerful and specific tool for the detection of disseminated cancer cells without false-positive results. Furthermore, it delivers enough cells for subsequent investigations such as characterization studies.