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Abstract
The purpose of this research project was to examine the capacity of anti-idiotypic antibodies and anti-idiotypic-like antibodies to regulate normal and malignant B-cell populations. Out of this project stemmed two new findings. The first was a technique for the rapid comparison of the primary amino acid sequences of antibody molecules. This method was applied to studying a panel of antibodies derived from an immune response characterized by a dominant idiotype where it was able to quickly distinguish between idiotype positive antibodies that differed by only a few amino acids. The technique was also applied to the analysis of monoclonal antibodies derived from fusions between human B-cell tumor containing tissue and mouse myeloma cells in order to identify which hybridomas were derived from the malignant B-cell population. The second finding involved techniques for growing non-murine antibody secreting hybridomas in immunosuppressed mice. Human x mouse and rat x mouse heterohybridomas grew readily in mice treated with cortisone and radiation but completely xenogeneic (rat x rat) hybrids did not. This was overcome by developing a ouabain resistant variant of the SP-2 mouse myeloma cell line and fusing it to the (rat x rat) hybridomas forming a "second generation" hybridoma cell lines that grew readily in immunosuppressed mice.
As an outgrowth of the human anti-idiotype project came an idiotype-like regulation model system. The idiotype-like regulation project was an attempt to extend the idiotype specific regulation effects of anti-idiotypic antibodies to the entire antigen binding antibody response. An antibody construct was made by covalently coupling antibodies specific for photoligand moiety to a peptide consisting of a photoligand attached to a hapten. The resulting complex had a single hapten specifically bound to each antigen binding region of the monoclonal antibody and was expected to influence antigen binding B-cells in much the same way as an anti-idiotypic antibody. However, when these complexes were injected into mice it was found that they did not modulate the subsequent antibody response. This lack of effect was due to a greatly decreased circulation time of the antibody-photoligand-hapten complex.
