HYBRIDS FROM NORMAL, GERM FREE, NUDE AND NEONATAL MICE PRODUCE MONOCLONAL AUTOANTIBODIES TO 8 DIFFERENT INTRACELLULAR STRUCTURES

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  • 1 January 1985
    • journal article
    • research article
    • Vol. 60  (2) , 417-426
Abstract
The supernatants of hybrids produced by fusion of splenocytes from normal, germ-free, nude and neonatal BALB c mice with mouse NS-1 myeloma cells were examined for immunofluorescence reactivity with viable and acetone fixed monolayers of syngeneic mouse fibroblasts and mouse 3T3 cells and with viable cell suspensions of syngeneic mouse erythrocytes and thymocytes. The supernatants of 70 of 419 (16.7%) hybrids from normal mice, 87 of 627 (13.9%) from germ-free mice, 28 of 240 (11.7%) from nude mice and 42 of 1020 (4%) from neonatal mice reacted with 8 different intracellular structures in mouse fibroblasts and 3T3 cells, and with rat and human fibroblasts. The reactive intracellular structures comprised stress fibers, intermediate filaments, cell membrane associated, Golgi complex, cytoplasm, cytoplasmic vesicles, mitochondria and nuclei. Of 45 stable clones derived by limiting dilution, 43 produced IgM antibodies and 2 IgG2b antibodies. Only 1 of the 2306 (0.04%) hybrids produced an autoantibody to the surface membrane of mouse thymocytes. These results show that B cells with reactivity towards different intracellular structures are present in the normal B cell repertoire whereas surface reactive B cells may be deleted or more profoundly suppressed or anergic and unable to form viable Ig producing hybridomas.