Burrone O R, Calabi F, Kefford R F, Milstein C
MRC Laboratory of Molecular Biology, Cambridge, UK.
EMBO J. 1983;2(9):1591-5. doi: 10.1002/j.1460-2075.1983.tb01629.x.
Somatic variants with constitutive changes in the expression of the cortical thymocyte differentiation antigen HTA 1 were derived from the T-cell leukemia line Molt 4 using monoclonal antibody NA1/34 and the fluorescent activated cell sorter. Cells with the highest and lowest fluorescence were sorted and expanded. After several cycles, high expressor variants, with 10- to 12-fold increased surface HTA 1 and low expressors, with a level of one third relative to the wild-type, were obtained. The stability of the high expressors was improved by cloning. In spite of the large differences in the level of HTA 1 between the various mutants and the wild-type, the expression of HLA-A,B,C or its increase following interferon-alpha stimulation, remained unchanged. Although in HTA 1 there was only a trace of beta2-microglobulin (beta2m) detected by lactoperoxidase labelling of the high expressor variants, significant levels of both beta2m and HTA 1 light chain beta(t) were observed in gels stained with Coomassie blue. The physiological association of beta(t) with HTA 1 was confirmed by the substantial increase of beta(t) which parallels the increase of HTA 1.
利用单克隆抗体NA1/34和荧光激活细胞分选仪,从T细胞白血病系Molt 4中获得了皮质胸腺细胞分化抗原HTA 1表达发生组成性变化的体细胞变体。对荧光强度最高和最低的细胞进行分选和扩增。经过几个循环后,获得了高表达变体,其表面HTA 1增加了10至12倍,以及低表达变体,其水平相对于野生型为三分之一。通过克隆提高了高表达变体的稳定性。尽管各种突变体与野生型之间HTA 1水平存在很大差异,但HLA - A、B、C的表达或其在α干扰素刺激后的增加保持不变。虽然在用高表达变体的乳过氧化物酶标记中仅检测到痕量的β2 - 微球蛋白(β2m),但在考马斯亮蓝染色的凝胶中观察到了显著水平的β2m和HTA 1轻链β(t)。β(t)与HTA 1的生理关联通过β(t)与HTA 1增加平行的大幅增加得到证实。