Breast cancer cells are effectively purged from peripheral blood progenitor cells with an immunomagnetic technique.

Peripheral blood progenitor cells (PBPCs) are being used increasingly to provide hematopoietic support after intensive chemotherapy. However, many investigators have detected tumor cells contaminating PBPC collections. Methods that eliminate the tumor cells and spare the normal hematopoietic progenitor cells may improve the number of long-term, disease-free survivors after intensive chemotherapy. We developed an effective method using anti-breast cancer murine monoclonal antibodies (MoAbs) and immunomagnetic beads to eliminate a low percentage of breast cancer cells from PBPCs. We identified optimal anti-breast cancer MoAbs that react with membrane glycoproteins and conditions for selective removal of tumor cells. Using three anti-breast cancer MoAbs (260F9, 317G5, and 520C9) at 0.8 microgram/ml, a cell concentration of 2 x 10(8) cells/ml and a bead:total cell ratio of 0.75 beads:1 cell, we eliminated 3.3-4.8 (mean, 4.1) logs of tumor cells consistently from a model system with 1% breast cancer cells and 99% normal PBPCs. Similar levels of tumor cell elimination were obtained with three breast cancer cell lines. Colony-forming units were not affected adversely, with a mean recovery of 200% compared with the control. A clinical trial has begun that uses immunomagnetically purged, autologous bone marrow and PBPCs to support patients with metastatic breast cancer receiving high-dose chemotherapy.

[1]  N. Davidson,et al.  Absence of breast cancer cells in a single-day peripheral blood progenitor cell collection after priming with cyclophosphamide and granulocyte-macrophage colony-stimulating factor. , 1995, Blood.

[2]  J. Gribben,et al.  Detection of minimal residual disease in patients with lymphomas using the polymerase chain reaction. , 1994, Important advances in oncology.

[3]  Rill,et al.  Direct demonstration that autologous bone marrow transplantation for solid tumors can return a multiplicity of tumorigenic cells. , 1994, Blood.

[4]  N. Davidson,et al.  Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques. , 1993, Blood.

[5]  G. Rosner,et al.  Comparative effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) on priming peripheral blood progenitor cells for use with autologous bone marrow after high-dose chemotherapy. , 1993, Blood.

[6]  D. Crowther,et al.  Peripheral blood progenitor cell transplantation in lymphoma and leukemia using a single apheresis. , 1993, Blood.

[7]  R. M. Fox,et al.  Effect of peripheral-blood progenitor cells mobilised by filgrastim (G-CSF) on platelet recovery after high-dose chemotherapy , 1992, The Lancet.

[8]  Gorin Nc Marrow purging: present status and future perspectives--efficacy in AML. , 1992 .

[9]  R. Bast,et al.  Elimination of clonogenic breast cancer cells from human bone marrow. A comparison of immunotoxin treatment with chemoimmunoseparation using 4‐hydroperoxycyclophosphamide, monoclonal antibodies, and magnetic microspheres , 1991, Cancer.

[10]  J. Vredenburgh,et al.  Reactivity of anti-CD15 monoclonal antibody PM-81 with breast cancer and elimination of breast cancer cells from human bone marrow by PM-81 and immunomagnetic beads. , 1991, Cancer research.

[11]  J. Vredenburgh,et al.  Elimination of small cell carcinoma of the lung from human bone marrow by monoclonal antibodies and immunomagnetic beads. , 1990, Cancer research.

[12]  R. Bast,et al.  Detection of breast carcinoma cells in human bone marrow using fluorescence-activated cell sorting and conventional cytology. , 1990, American journal of clinical pathology.

[13]  D. Hayes,et al.  Evaluation of monoclonal antibody DF3 conjugated with ricin as a specific immunotoxin for in vitro purging of human bone marrow. , 1990, Cancer research.

[14]  R. Bast,et al.  Elimination of malignant clonogenic breast cancer cells from human bone marrow. , 1989, Cancer research.

[15]  R. Bast,et al.  Heterogeneity of antigen expression in benign and malignant breast and ovarian epithelial cells , 1989, International journal of cancer.

[16]  S. Ménard,et al.  Monoclonal antibody detection of carcinoma cells in bone marrow biopsy specimens from breast cancer patients , 1988, Cancer.

[17]  S. Funderud,et al.  Monosized magnetic particles and their use in selective cell separation , 1988 .

[18]  R. Gale,et al.  Advanced breast cancer: high-dose chemotherapy and bone marrow autotransplants. , 1988, Annals of internal medicine.

[19]  D. Weisenburger,et al.  Autologous peripheral hematopoietic stem cell transplantation restores hematopoietic function following marrow ablative therapy. , 1988, Blood.

[20]  L. To,et al.  Early lympho-hemopoietic recovery after autografting using peripheral blood stem cells in acute non-lymphoblastic leukemia. , 1988, Transplantation proceedings.

[21]  L. Gordon,et al.  Autologous bone marrow transplantation in breast cancer: separation of clonogenic tumor cell colonies by gradient fractionation. , 1988, Experimental hematology.

[22]  B. Dörken,et al.  Immunomagnetic removal of B-lymphoma cells from human bone marrow: a procedure for clinical use. , 1988, Bone marrow transplantation.

[23]  R. Bast,et al.  Combined chemoseparation and immunoseparation of clonogenic T lymphoma cells from human bone marrow using 2'-deoxycoformycin, deoxyadenosine, 3A1 monoclonal antibody, and complement. , 1987, Cancer research.

[24]  C. Patte,et al.  Repeated high-dose chemotherapy followed by purged autologous bone marrow transplantation as consolidation therapy in metastatic neuroblastoma. , 1987, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[25]  S. Margel,et al.  Purging breast cancer cells in preparation for autologous bone marrow transplantation. , 1987, Bone marrow transplantation.

[26]  V. Bosnes,et al.  DEPLETION OF T LYMPHOCYTES FROM HUMAN BONE MARROW: Use of Magnetic Monosized Polymer Microspheres Coated with T‐Lymphocyte‐Specific Monoclonal Antibodies , 1987, Transplantation.

[27]  R. Braylan,et al.  Selective loss of expression of a tumor-associated antigen on a human leukemia cell line induced by treatment with monoclonal antibody and complement. , 1987, Journal of the National Cancer Institute.

[28]  A. Gee,et al.  Low antigen density tumor cells: an obstacle to effective autologous bone marrow purging? , 1987, Cancer investigation.

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

[30]  BONE-MARROW AUTOTRANSPLANTATION IN MAN Report of An International Cooperative Study , 1986, The Lancet.

[31]  T. Powles,et al.  In vitro and in vivo effects of a monoclonal antibody-toxin conjugate for use in autologous bone marrow transplantation for patients with breast cancer. , 1986, Cancer research.

[32]  R. Bast,et al.  Elimination of clonogenic tumor cells from human bone marrow using a combination of monoclonal antibody:ricin A chain conjugates. , 1986, Cancer research.

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

[34]  J. Graham-Pole,et al.  Antibody-complement killing of neuroblastoma cells. , 1985, Progress in clinical and biological research.

[35]  T. Powles,et al.  DETECTION OF MICROMETASTASES IN PATIENTS WITH PRIMARY BREAST CANCER , 1983, The Lancet.

[36]  R. Bast,et al.  Elimination of leukemic cells from human bone marrow using monoclonal antibody and complement. , 1983, Cancer research.

[37]  R. Bast,et al.  Elimination of leukemic cells from rat bone marrow using antibody and complement. , 1981, Cancer research.

[38]  E. Lasfargues,et al.  Isolation of two human tumor epithelial cell lines from solid breast carcinomas. , 1978, Journal of the National Cancer Institute.

[39]  J. Fogh,et al.  Absence of HeLa cell contamination in 169 cell lines derived from human tumors. , 1977, Journal of the National Cancer Institute.

[40]  J. Macnab Molecular Studies in Viral Neoplasia: A Collection of Papers Presented at the Twenty-Fifth Annual Symposium on Fundamental Cancer Research, 1972, at the University of Texas M. D. Anderson Hospital and Tumor Institute at Houston , 1975 .

[41]  J. Fogh,et al.  New Human Tumor Cell Lines , 1975 .

[42]  B. W. Brown,et al.  The Spearman Estimator for Serial Dilution Assays , 1961 .