Elimination of myeloma cells from bone marrow by using monoclonal antibodies and magnetic immunobeads.

The efficacy of immunomagnetic beads to purge human myeloma cells from bone marrow ex vivo was evaluated. The optimal conditions for purging were studied first by using three myeloma cell lines: RPMI-8226, SKO-007, and SKMM-2. Myeloma cells labeled with the vital fluorescent dye Hoechst 33342 were admixed with normal bone marrow cells, and two monoclonal antibodies reactive with the myeloma cells (PCA-1 and BL-3) were added alone or in combination with the cells. Magnetic beads coated with goat antimouse immunoglobulin G were then added, and the tumor cells to which beads were attached were separated from the mixture with a magnet. The efficacy of tumor cell removal was dependent on the bead-to-tumor ratio; a ratio of more than 500 was optimal in the presence of excess normal marrow cells. The combination of monoclonal antibodies PCA-1 and BL-3 increased the tumor cell removal as compared with either antibody alone. Two cycles of treatment were more effective than one cycle was. Under optimal conditions, 2.3 to 4 logs of tumor cells could be removed from the mixture containing 10% myeloma cells without a significant loss of normal hematopoietic progenitors as measured by CFU-GM, CFU-GEM, and BFU-E. When the efficacy of this procedure was tested on fresh bone marrow from patients with multiple myeloma (MM) by using the combination of PCA-1, BL-3, and J-5, 1.6 to 2.5 logs of tumor cells could be removed by one cycle of treatment, even from marrows containing less than 10% myeloma cells. These observations support the use of monoclonal antibody combinations and immunobeads as a reliable and nontoxic method to eliminate contaminating myeloma cells ex vivo in preparation for autologous bone marrow transplantation in patients with MM.

[1]  M. Slevin,et al.  Multiple myeloma treated with high dose intravenous melphalan , 1987, British journal of haematology.

[2]  J. Gerdes,et al.  Novel type of proliferating lymphoplasmacytoid cell with a characteristic spotted immunofluorescence pattern. , 1987, The Journal of clinical investigation.

[3]  J. Gabrilove,et al.  Activities of four purified growth factors on highly enriched human hematopoietic progenitor cells. , 1987, Blood.

[4]  O. Fodstad,et al.  Elimination of B-lymphoma cells from human bone marrow: model experiments using monodisperse magnetic particles coated with primary monoclonal antibodies. , 1987, Cancer research.

[5]  J. Kersey,et al.  Marrow purging in autologous bone marrow transplantation for T-lineage acute lymphoblastic leukemia: efficacy of ex vivo treatment with immunotoxins and 4-hydroperoxycyclophosphamide against fresh leukemic marrow progenitor cells. , 1987, Blood.

[6]  M. Stone,et al.  Characterization of a monoclonal antibody having selective reactivity with normal and neoplastic plasma cells. , 1987, Blood.

[7]  R. Seeger,et al.  Model system for removing neuroblastoma cells from bone marrow using monoclonal antibodies and magnetic immunobeads. , 1986, Cancer research.

[8]  Zander,et al.  High-dose melphalan with autologous bone marrow transplantation for multiple myeloma. , 1986, Blood.

[9]  J. Katzmann,et al.  A monoclonal antibody reactive with a subset of human plasma cells , 1986, British journal of haematology.

[10]  G. Spitzer,et al.  Clinical studies of autografting in acute lymphocytic leukaemia. , 1986, Clinics in haematology.

[11]  H. Zola,et al.  Immunological phenotype of neoplasms involving the B cell in the last step of differentiation , 1986, British journal of haematology.

[12]  D. Campana,et al.  Identification of malignant plasma cell precursors in the bone marrow of multiple myeloma. , 1985, The Journal of clinical investigation.

[13]  T. Grogan,et al.  CALLA-positive myeloma: an aggressive subtype with poor survival. , 1985, Blood.

[14]  J. Leonard,et al.  Preclinical studies on the use of selective antibody-ricin conjugates in autologous bone marrow transplantation. , 1985, Blood.

[15]  R. Bast,et al.  Elimination of clonogenic Burkitt's lymphoma cells from human bone marrow using 4-hydroperoxycyclophosphamide in combination with monoclonal antibodies and complement. , 1985, Blood.

[16]  T. Lebien,et al.  Utilization of a colony assay to assess the variables influencing elimination of leukemic cells from human bone marrow with monoclonal antibodies and complement. , 1985, Blood.

[17]  D. Bergsagel.  Controversies in the treatment of plasma cell myeloma. , 1985, Postgraduate medical journal.

[18]  W. O'Fallon,et al.  Multiple myeloma: significance of plasmablastic subtype in morphological classification. , 1985, Blood.

[19]  R. Bast,et al.  Elimination of malignant clonogenic cells from human bone marrow using multiple monoclonal antibodies and complement. , 1985, Cancer research.

[20]  C. Y. Wang,et al.  Preparation and characterization of monoclonal antibodies recognizing three distinct differentiation antigens (BL1, BL2, BL3) on human B lymphocytes. , 1984, Journal of immunology.

[21]  L. Nadler,et al.  A monoclonal antibody with reactivity restricted to normal and neoplastic plasma cells. , 1984, Journal of immunology.

[22]  M. Greaves,et al.  An immunotoxin with therapeutic potential in T cell leukemia: WT1-ricin A , 1984 .

[23]  J. Kemshead,et al.  REMOVAL OF NEUROBLASTOMA CELLS FROM BONE MARROW WITH MONOCLONAL ANTIBODIES CONJUGATED TO MAGNETIC MICROSPHERES , 1984, The Lancet.

[24]  L. Nadler,et al.  Antigens on human plasma cells identified by monoclonal antibodies. , 1983, Journal of immunology.

[25]  M. Körbling,et al.  4‐Hydroperoxyclophosphamide: a model for eliminating residual human tumour cells and T‐lymphocytes from the bone marrow graft , 1982, British journal of haematology.

[26]  R. Storb,et al.  Identical twin marrow transplantation in multiple myeloma. , 1982, Acta haematologica.

[27]  A. Houghton,et al.  Heterogeneity in surface antigen and glycoprotein expression of cell lines derived from different melanoma metastases of the same patient. Implications for the study of tumor antigens , 1981, The Journal of experimental medicine.

[28]  R. Bast,et al.  Absence of common ALL antigen on normal bipotent myeloid, erythroid, and granulocyte progenitors. , 1981, Blood.

[29]  H. Kaplan,et al.  Human-human hybridomas producing monoclonal antibodies of predefined antigenic specificity. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[30]  S. Salmon,et al.  Pretreatment tumor mass, cell kinetics, and prognosis in multiple myeloma. , 1980, Blood.

[31]  H. Lazarus,et al.  A monoclonal antibody to human acute lymphoblastic leukaemia antigen , 1980, Nature.

[32]  M. Greaves,et al.  Acute lymphoblastic leukaemia associated antigen. IV. Expression on non-leukaemic 'lymphoid' cells. , 1980, Leukemia research.

[33]  T. Lister,et al.  Plasma cell leukemia (PCL): A report on 15 patients. , 1978, Blood.

[34]  B. Clarkson,et al.  Improved survival times in multiple myeloma treated with melphalan, prednisone, cyclophosphamide, vincristine and BCNU: M-2 protocol. , 1977, The American journal of medicine.

[35]  B. Clarkson,et al.  Immunoglobulin synthesis by human reticulum sarcoma cells in vivo and during long-term culture in vitro. , 1969, Cancer research.

[36]  G. Moore,et al.  Production of Free Light Chains of Immunoglobulin by a Hematopoietic Cell Line Derived from a Patient with Multiple Myeloma.∗ , 1967, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.