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Elimination of Malignant Clonogenic Cells from Human Bone Marrow Using Multiple Monoclonal Antibodies and Complement¹

Divisions of Tumor Immunology [R.C.B., P. D. F., C. M., L. N., J. R.], Medicine [R. C. B.], Biostatistics [R. G.], and Pediatric Oncology [J. L., S. S.], Dana-Farber Cancer Institute; Department of Medicine [R. C. B., L. N., J. R.], Brigham and Women's Hospital; Department of Hematology and Oncology [J. L., S. S.], Children's Hospital Medical Center; Department of Biostatistics [R. G.], Harvard School of Public Health; and the Departments of Medicine [R. C. B., P. D. F., L. N., J. R.] and Pediatrics [J. L., S. S.], Harvard Medical School, Boston, Massachusetts 02115

A clonogenic assay has been developed that utilizes Burkitt's lymphoma tumor cell lines to detect elimination of up to 5 logs of tumor cell contamination within human bone marrow. Different Burkitt's lymphoma lines bear one or more of a group of markers, including common acute lymphoblastic leukemia antigen gp26 (glycoprotein with a molecular weight of 26,000), B1, surface membrane immunoglobulin, HLA, ß₂-microglobulin, and la. Burkitt's tumor cells of the Namalwa line have been mixed with a 20-fold excess of irradiated human bone marrow cells. After treatment with one or more monoclonal antibodies and rabbit complement (RC), mixtures have been grown on a monolayer of irradiated human bone marrow cells and tumor cells enumerated by limiting dilution. Multiple treatments with antibody and RC were more effective than a single treatment in destroying clonogenic tumor cells which bore relevant determinants. Human serum components inhibited the lytic activity of RC in the presence of murine monoclonal antibodies. The total concentration of bone marrow cells proved critical in determining the complete elimination of tumor. Incubation of the Namalwa tumor cell line with RC and the J2 anti-gp26 eliminated more than 3 logs of malignant cells from a 20-fold excess of human bone marrow. Combinations of two monoclonal antibodies were more effective than any single antibody in eliminating Namalwa cells. A combination of three monoclonal reagents was no more effective than a combination of J2 and B1 or J2 and J5 in eliminating Namalwa cells. Treatment of human bone marrow with three antibodies and RC did not, however, produce a selective loss of nonmalignant GM-CFU-C, CFU-E, or BFU-E.

¹ Supported in part by National Cancer Institute Grants CA28740 and CA19589.

² Scholar of the Leukemia Society of America. To whom requests for reprints should be addressed, at Box 3843, Duke University Medical Center, Durham, NC 27710.

³ Supported by the American-Italian Cancer Research Foundation and the Associazione Italiana per la Ricerca sul Cancro.

⁴ Fellow of the Dyson Foundation.

⁵ Scholar of the Leukemia Society of America.

Received 1/23/84. Accepted 10/16/84.

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