McCubrey J, McKearn J P, Köhler G
Eur J Immunol. 1985 Nov;15(11):1117-24. doi: 10.1002/eji.1830151110.
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.
已将来自2,4,6-三硝基苯基(TNP)特异性杂交瘤Sp6的重排μ和κ基因导入处于三个不同分化阶段的B细胞以及5种非B细胞系中,以确定免疫球蛋白(Ig)基因表达的水平和模式。在用μ和κ基因转化的前B细胞中,产生了低水平的Sp6特异性μRNA,与杂交瘤Sp6相比,产生的μ蛋白减少了约210倍,κ蛋白减少了约800倍。Ig蛋白存在于细胞内,但在细胞膜上未检测到。在成熟的表面sIg + B细胞转化体中,产生的μSp6和κSp6蛋白及RNA水平高于前B细胞转化体(μ为12倍,κ为70倍)。这些转化体在细胞膜上展示了μSp6和κSp6蛋白,并且还分泌了转染的Ig产物。浆细胞转化体产生的μSp6和κSp6蛋白量最高。这些转化体分泌五聚体IgM,但在细胞膜上未显示出可检测到的这些蛋白量。T细胞和成纤维细胞转化体产生正常大小的μSp6和κSp6蛋白形式的Ig。所有其他μSp6和κSp6非B细胞转化体(黑色素瘤、畸胎瘤和巨噬细胞)未能产生足够的Ig来确定Ig蛋白的分子量是否正确。产生Ig蛋白的T细胞和成纤维细胞转化体未在细胞膜上分泌或显示可检测到的Ig。Ig的表达并未抑制T细胞转化体中T细胞抗原Thy-1的表达。