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Monoclonal Antibody-defined Correlations in Melanoma between Levels of G_D2 and G_D3 Antigens and Antibody-mediated Cytotoxicity¹

The Wistar Institute, Philadelphia, PA 19104 [J. T., M. T., Y. K., M. H., M. D. L., M. S., Z. S., H. K.]; The Pigmented Lesion Study Group, Comprehensive Cancer Center, Department of Pathology, University of Pennsylvania, Philadelphia, PA 19104 [D. E. E., W. M. C.]; and the Department of Medical Biochemistry, University of Göteborg, Göteborg, Sweden [K-A. K.]

A monoclonal antibody is described that specifically detects the ganglioside antigens G_D2 and G_D3, binding preferentially to G_D2, in melanoma. Antibody specificity was demonstrated with solid-phase radioimmunoassay and enzyme-linked immunosorbent assay as well as by immunostaining on thin-layer chromatography plates using structurally characterized gangliosides. Binding of both the IgG3 antibody and its IgG2a switch variant were assayed on live cells by cytofluorography and by immunoperoxidase staining on frozen tissue sections. The binding patterns correlated with antitumor activity in antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity assays with human effector cells and complement in an ¹¹¹In-release assay using cell lines derived from the same individual. The significant level of killing in all tumor cells tested that express G_D2, G_D3, or both, suggests the importance of multiple specificity towards tumor antigens, i.e., binding of a monoclonal antibody to two or more tumor-associated antigens.

¹ This work was supported by Grant 3967 from the Swedish Medical Research Council and by Grants CA-25874, CA-21124, and CA-10815 from the National Institutes of Health.

Received 8/25/86. Revised 11/19/86. Accepted 11/25/86.