Storb U, Hager L, Putnam D, Buck L, Farin F, Clagett J
Proc Natl Acad Sci U S A. 1976 Jul;73(7):2467-71. doi: 10.1073/pnas.73.7.2467.
We investigated by molecular hybridization whether T cells contain RNA sequences homologous to RNA which codes for immunoglobulin kappa-chain (k-chain). A radioactive probe of complementary DNA (cDNA) was prepared by transcription of purified k-chain mRNA from mouse myeloma MOPC-41 with reverse transcriptase (RNA-dependent-DNA nucleotidyltransferase) from avian myeloblastosis virus. The cDNA probably corresponded only to the constant region and 3'-terminus of k-chain mRNA. Kappa-chain cDNA was found to hybridize efficiently with RNA from both thymus cells and an established culture of thymoma cells. The thymus and thymoma cells contained 99.8% and 100% theta-positive cells, respectively. Quantitatively the average thymus T cell (thymus derived lymphocyte) contained about one half as much k-chain mRNA as the average spleen B cell ("bursa" dependent lymphocyte), whereas the thymoma cells contained only 1/33 as much. Control hybridizations of k-chain cDNA with myeloma and liver RNA support the conclusion that T cells in the thymus and in the thymoma cell line synthesize k-chain mRNA-like molecules. The thermal stability of hybrids of k-chain cDNA with RNA from spleen, thymus, thymoma, and another k-chain producing myeloma tumor was lower than that with MOPC-41 RNA. This finding may be due to the existence of several slightly different ck genes in the mouse as suggested by various control experiments.
我们通过分子杂交研究了T细胞是否含有与编码免疫球蛋白κ链(κ链)的RNA同源的RNA序列。通过用禽成髓细胞瘤病毒的逆转录酶(RNA依赖性DNA核苷酸转移酶)转录从小鼠骨髓瘤MOPC-41纯化的κ链mRNA来制备互补DNA(cDNA)放射性探针。该cDNA可能仅对应于κ链mRNA的恒定区和3'末端。发现κ链cDNA与胸腺细胞和已建立的胸腺瘤细胞培养物中的RNA有效杂交。胸腺和胸腺瘤细胞分别含有99.8%和100%的θ阳性细胞。定量地,平均胸腺T细胞(胸腺衍生淋巴细胞)所含的κ链mRNA约为平均脾B细胞(“法氏囊”依赖性淋巴细胞)的一半,而胸腺瘤细胞仅含其1/33。κ链cDNA与骨髓瘤和肝脏RNA的对照杂交支持以下结论:胸腺和胸腺瘤细胞系中的T细胞合成κ链mRNA样分子。κ链cDNA与来自脾脏、胸腺、胸腺瘤和另一种产生κ链的骨髓瘤肿瘤的RNA的杂交体的热稳定性低于与MOPC-41 RNA的杂交体。如各种对照实验所表明的,这一发现可能是由于小鼠中存在几个略有不同的ck基因。