Transformation of B and non-B cell lines with the 2,4,6,-trinitrophenyl (TNP)-specific immunoglobulin genes.
The rearranged mu and kappa genes from the 2,4,6-trinitrophenyl (TNP)-specific hybridoma Sp6 have been introduced into B cells from three different stages of differentiation as well as 5 non-B cell lines to determine the levels and modes of immunoglobulin (Ig) gene expression. In pre-B cells transformed with the mu and kappa genes, low levels of Sp6-specific mu RNA were produced and approximately 210-fold less mu and 800-fold less kappa proteins were produced than in the hybridoma Sp6. The Ig proteins were present intracellularly, but were not detected on the cell membrane. In mature surface sIg+ B cell transformants, higher levels of mu Sp6 and kappa Sp6 proteins and RNA were produced than in the pre-B cell transformants (12 X mu, 70 X kappa). These transformants displayed the mu Sp6 and kappa Sp6 proteins on the cell membrane and also secreted the transfected Ig product. Plasma cell transformants produced the highest amounts of mu Sp6 and kappa Sp6 proteins. These transformants secreted pentameric IgM but did not display detectable amounts of these proteins on the cell membrane. T cell and one fibroblast transformant produced Ig as normal sized mu Sp6 and kappa Sp6 proteins. All other mu Sp6 and kappa Sp6 non-B cell transformants (melanoma, teratoma and macrophage) failed to produce enough Ig to determine whether the Ig proteins were of the correct molecular weights. The T cell and fibroblast transformants that produced Ig proteins did not secrete or display detectable Ig on the cell membrane. The expression of Ig did not inhibit the expression of the T cell antigen Thy-1 in the T cell transformants.
McCubrey
J
J
McKearn
J P
JP
Köhler
G
G
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D002460:Cell Line; D004247:DNA / Q000032:analysis; D005347:Fibroblasts / Q000378:metabolism; D005796:Genes; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D007136:Immunoglobulins / Q000032:analysis / Q000235:genetics*; D008213:Lymphocyte Activation; D008264:Macrophages / Q000378:metabolism; D008545:Melanoma / Q000235:genetics; D051379:Mice; D009578:Nitrobenzenes / Q000276:immunology*; D010950:Plasma Cells / Q000378:metabolism; D012313:RNA / Q000032:analysis; D051381:Rats; D013234:Stem Cells / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism; D013724:Teratoma / Q000235:genetics; D014162:Transfection; D014301:Trinitrobenzenes / Q000276:immunology*
D007145:Immunoglobulin kappa-Chains; D007148:Immunoglobulin mu-Chains; D007136:Immunoglobulins; D009578:Nitrobenzenes; D014301:Trinitrobenzenes; D012313:RNA; D004247:DNA