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In Vitro Bone Marrow Purging of Multidrug-resistant Cells with a Mouse Monoclonal Antibody Directed against M_r 170,000 Glycoprotein and a Saporin-conjugated Anti-Mouse Antibody¹

Istituto di Ematologia "L. & A. Seragnoli" [A. D., P. L. T., G. V., G. V., M. G., A. B., C. T., S. P., S. T.] and Dipartimento di Patologia Sperimentale [A. B., F. S.], Universita' di Bologna, Bologna, Italy; Cattedra di Ematologia [M. M., R. F., M. B.] and Istituto di Scienze Mediche [D. D.], Universita' di Udine, Udine, Italy; Farmitalia Carlo Erba Research Center, Nerviano, Italy [M. G.]; and Division of Experimental Chemotherapy, Japanese Foundation for Cancer Research, Tokyo, Japan [T. T.]

Selective elimination f multidrug resistance-positive cells (LoVo/Dx) was obtained by using the monoclonal antibody MRK 16, which recognizes a surface epitope of the M_r 170,000 glycoprotein, and a sheep antimouse immunoglobulin antibody, conjugated to the ribosome-inactivating protein saporin 6. The killing was greatly decreased or even abolished by adding the monoclonal antibody at a 100-fold concentration. Both the MRK 16 and anti-mouse saporin 6 conjugate did not show any killing activity when they were used separately. In cell suspensions composed of 90% normal bone marrow cells and 10% multidrug resistance-positive cells, the monoclonal antibody MRK 16 followed by the anti-mouse immunotoxin caused the elimination of 99% multidrug resistance-positive cells, as revealed by immunofluorescence and immunocytochemistry as well as by a clonal assay. Human normal hematopoietic precursors (granulomonocytic colony-forming units, erythroid burst-forming units, and multipotent granulomonocytic, erythroid, and megakaryocytic-forming units) were not affected by the MRK 16 plus immunotoxin treatment. This technique might be suitable for ex vivo bone purging in an appropriate clinical setting, such as autologous bone marrow transplantation.

¹ Supported by Regione Emilia Romagna, Delibera n. 1970 del 13/5/86, CNR finalized project "Oncologia," and by Associazione Italiana per la Ricerca sul Cancro.

² Supported by a grant from Associazione Italiana per la Ricerca sul Cancro (AIRC), Milano, Italy.

³ To whom requests for reprints should be addressed, at Istituto di Ematologia "L. & A. Seragnoli," Via Massarenti 9, 40138 Bologna, Italy.

Received 6/29/89. Revised 1/ 4/90.

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