A new method for the detection of cell surface immunoglobulin labeled with isotopic precursors is described. The method consists of the aggregation of surface Ig on cells with specific antibody (heterologous) and the subsequent removal of antigen-antibody complexes by the combination of high speed centrifugation and immunoprecipitation of remaining soluble complexes using antibody to the heterologous Ig. Using this method, the kinetics of appearance of cell surface Ig and its turnover were studied in murine splenocytes. The results suggest that cell surface Ig is synthesized and transported in the same manner as secretory Ig rather than being synthesized on the plasma membrane. The turnover of intracellular and cell surface Ig in lymphocytes is slow. In contrast, intracellular Ig in plasma cells is rapidly secreted and usually without a cell surface phase. Cell surface Ig was shown to be radiolabeled with [3H]glucosamine, -galactose, and -fucose. The proportion of cell surface to intracellular (nonsurface) Ig labeled with these precursors suggests the same sequence of addition of sugars to Ig destined to be on the surface of lymphocytes as with Ig which will be secreted by plasma cells. Results with this new method also confirm earlier conclusions based on experiments using cell surface iodination: 8S IgM is the predominant Ig on the surface of murine splenocytes and the molecule appears to be attached by its µ-chains.
CELL SURFACE IMMUNOGLOBULIN : IX. A NEW METHOD FOR THE STUDY OF SYNTHESIS, INTRACELLULAR TRANSPORT, AND EXTERIORIZATION IN MURINE SPLENOCYTES
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Ellen S. Vitetta, Jonathan W. Uhr; CELL SURFACE IMMUNOGLOBULIN : IX. A NEW METHOD FOR THE STUDY OF SYNTHESIS, INTRACELLULAR TRANSPORT, AND EXTERIORIZATION IN MURINE SPLENOCYTES . J Exp Med 1 June 1974; 139 (6): 1599–1620. doi: https://doi.org/10.1084/jem.139.6.1599
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