USE OF MONOCLONAL ANTIBODIES SPECIFIC FOR NEUROBLASTOMA (NB) IN THE DETECTION AND COMPLEMENT MEDIATED LYSIS OF MICROSCOPIC DISEASE

Abstract

Three murine hybridomas [3A7(IgM), 3G6(IgM), 3F8(IgG₃)] against human NB cell surface antigens were established using standard hybridization techniques. Both by ELISA and immunofluorescence, these monoclonal antibodies do not bind to normal bone marrow (BM) or peripheral blood cells. They all bind intensely to human NB and NB cell lines (except SKNSH and SKNMC). The cross reactivity patterns with other sarcomas and their relative degree of binding suggest that the three monoclonal antibodies have different specificities and are not reacting with common neuro-ectodermal antigens. Using immunofluorescence these antibodies can detect reproducibly ≤0.1% of tumor cells seeded in BM samples. Patients with widespread NB even though negative in their BM aspirate and biopsy by standard light microscopy are often positive by immunofluorescence in their BM aspirates with all 3 monoclonals. Less than 250 ng of 3A7 or 3G6 can kill 100% of 3X10⁵ NB cells in the presence of 1:8 dilution of guinea pig complement, while not affecting normal blood or BM cells. In summary (1) these antibodies appear to be useful in detecting and monitoring residual NB in BM samples, (2) they are potent cytotoxic antibodies and may be suitable for complete and specific removal of residual NB cells before autologous BM transplantation.

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