Surface immunoglobulin of guinea-pig leukaemic lymphocytes.
The surface immunoglobulin of the transplantable L2C leukaemia of strain 2 guinea-pigs has been investigated. The immunoglobulin is seen to be synthesized when the cells are maintained in culture, indicating its intrinsic origin. Immunolabelling of the cell surface and immunochemical study of the Fab released by limited surface proteolysis indicate the presence of immunoglobulin of class IgM. IgG and free light chains were not detected, and there is unlikely to be an appreciable amount of immunoglobulin of any other class. The amount of immunoglobulin present, in terms of 4-chain monomers, is approximately 100,000 molecules per cell. Its half-life, calculated from the rate of reappearance in vitro of surface Fab after proteolytic clearing, is approximately 5 hours. Immunoglobulin secreted into the environment appears to arise predominantly or entirely from the cell surface: there is no evidence of an appreciable export of immunoglobulin which does not have a surface phase. Papain at 0.06 mg/ml rapidly removes the surface Fab. Residual Fcmu can then be detected by immunofluorescence, suggesting that papain cleaves surface IgM at a hinge region with the molecule in situ on the membrane. The released Fab is only moderately susceptible to degradation by papain at the enzyme: substrate ratio prevailing. It has been possible to isolate it from the papain digest by immuno-adsorption, with a notional yield of 75 mug per 10-10 cells, and then to prepare antisera against it.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000032:analysis; D000917:Antibody Formation; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002462:Cell Membrane / Q000276:immunology; D002478:Cells, Cultured; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D007140:Immunoglobulin Fab Fragments / Q000032:analysis; D007128:Immunoglobulin Fragments / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis*; D007457:Iodine Radioisotopes; D007942:Leukemia, Experimental / Q000276:immunology*; D008214:Lymphocytes / Q000276:immunology*; D008968:Molecular Conformation; D009368:Neoplasm Transplantation; D010206:Papain; D011863:Radioimmunoassay; D011947:Receptors, Antigen, B-Cell
D000888:Antibodies, Anti-Idiotypic; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D007075:Immunoglobulin M; D007457:Iodine Radioisotopes; D011947:Receptors, Antigen, B-Cell; D010206:Papain