A Study of Cell-Surface Characteristics of Macrophage Hybridomas and How tthey are Affected by Activation of the Macrophage Hybridomas with Gamma-Interferon
A series of macrophage hybridomas were geerated by fusion of splenic adherent cells with macrophage P388Dl tumor cells. The production of such a series of macrophage clones is of great assistance in analyzing macrophage biology and dissecting structural and functional relationships since they possess the advantage of maintaining stable phenotypic markers and functional properties during continuous in vitro culture, whereas in a live mouse relatively few macrophage cells can maintain such stable properties. Qualitative and quantitative differences in phenotype and functional activity were noted within this series of macrophage hybridomas. A sample from the forty-two clones originally produced was selected for initial analysis; this sample of clones was selected because they illustrated the heterogeneity noted within the entire panel. To characterize the surface markers on each of the macrophage cell lines, certain monoclonal antibodies of known specificity were added to each cell line using the technique of radioimmunoassay. The macrophage cell lines were then activated using a lymphokine, gamma-interferon, and the monoclonal antibodies were added to the activated hybridomas to observe any changes in their cell surface characteristics as a result of activation. Time and dose studies were performed to determine the optimum time and concentration of gammainterferon to be used to activate the macrophage hybridomas.
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