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Sensitivity of Immunocytochemical Detection of Breast Cancer Cells in Human Bone Marrow¹

Breast Cancer Research Laboratory, Department of Surgery [M. P. O., S. A.], Department of Epidemiology and Biostatistics [G. Y. W.], Human Tumor Immunology Laboratory [L. J. O.], and Department of Pathology [R. J. C., P. P. R.], Memorial Sloan-Kettering Cancer Center, New York, New York 10021

We have previously shown that occult micrometastases can be detected in the bone marrow of breast cancer patients, at the time of initial treatment, using a panel of epithelial specific monoclonal antibodies indirectly labeled with fluorescein. These monoclonal antibodies permit us to detect cancer cells at a concentration of two/million normal bone marrow cells. Immunofluorescence carries the disadvantage that detailed morphological examination of detected cells cannot be accomplished. A modification of the alkaline phosphatase anti-alkaline phosphatase method has been used to detect cancer cells and to observe their morphology in human bone marrow. The sensitivity of this method has been examined using an established human metastatic breast cancer cell line (MCF-7) mixed with normal bone marrow cells at various dilutions from 400 cancer cells/10⁶ marrow cells to 10 cancer cells/10⁶ marrow cells. The number of immunocytochemically stained MCF-7 cells counted at each concentration was related to the concentration by a simple nonlinear statistical model. At a concentration of 10 cancer cells/10⁶ bone marrow cells, the model shows that this method has the sensitivity to detect between four and six MCF-7 cells 95% of the time. Extrapolation, using this model, predicts that at the very low concentration of one cancer cell/10⁶ marrow cells, there is a 95% chance of detecting the cancer cell. This assay may be a very sensitive method for detecting cancer cells in the bone marrow of breast cancer patients.

¹ This project was supported by an American Cancer Society Clinical Research Award (PDT-367), the Society of Sloan-Kettering, the Charles and Helen Lazarus Charitable Foundation, and the Iris and B. Gerald Cantor Foundation.

² To whom requests for reprints should be addressed, at Breast Cancer Research Laboratory, Department of Surgery, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021.

Received 5/21/90. Accepted 3/ 1/91.

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