Eradication of Neuroblastoma Cells in Vitro by Monoclonal Antibody and Human Complement: Method for Purging Autologous Bone Marrow

Ulla M. Saarinen, Peter F. Coccia, Nai Kong V. Cheung, Stanton L. Gerson, Robert Pelley

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Abstract

We describe an in vitro method which is useful for purging autologous bone marrow of neuroblastoma cells. The method utilizes a single murine monoclonal antibody 3G6 (an immunoglobulin Mk) which we have previously developed against the ganglbside G02; undiluted human complement; and unfraction-ated whole bone marrow at 1 x 107 nucleated cells/ml. Tumor cell clonogenic assays, Hoechst 33342 fluorescent nuclear stain, and trypan blue viability stain methods were used to assay cytotoxicity. This complement-mediated cytotoxicity technique killed 99.9-100% of neuroblastoma cell lines NMB-7, LAN-1, LAN-5, and IMR-6, while normal marrow precursor cells were not detectably damaged. The presence of normal bone marrow did not inhibit the human complement-mediated cytotoxicity. Applying the cytotoxicity method to whole unseparated bone marrow demonstrated killing of seeded neuroblastoma cells, with no gross hemolysis or cell clumping. The method did not require expensive special equipment, use of animal complement sera, or prior fractionation of the bone marrow. The average marrow nucleated cell recovery was 95%. These studies indicate that in vitro purging of autologous marrow infiltrated with neuroblastoma with monoclonal antibody 3G6 and human complement is both technically feasible and effective in eradicating residual tumor while preserving bone marrow stem cells.

Original languageEnglish (US)
Pages (from-to)5969-5975
Number of pages7
JournalCancer Research
Volume45
Publication statusPublished - Nov 1 1985

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ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Saarinen, U. M., Coccia, P. F., Cheung, N. K. V., Gerson, S. L., & Pelley, R. (1985). Eradication of Neuroblastoma Cells in Vitro by Monoclonal Antibody and Human Complement: Method for Purging Autologous Bone Marrow. Cancer Research, 45, 5969-5975.