Kavathas P, Herzenberg L A
Proc Natl Acad Sci U S A. 1983 Jan;80(2):524-8. doi: 10.1073/pnas.80.2.524.
We isolated stable transformants of mouse L cells expressing human cell surface differentiation antigens by using immunofluorescence with monoclonal antibodies and selection with a fluorescence-activated cell sorter (FACS). Mouse L cells (TK-) were cotransformed with human cellular DNA and the herpes simplex virus thymidine kinase (TK) gene. TK+ transformants were first selected. The TK+ populations were stained with various fluorescent antibodies to membrane antigens, and positive cells were sorted and cloned by using a FACS. Transformants for HLA class I antigens, for beta 2-microglobulin, and for the T-cell differentiation antigens Leu-1 and Leu-2 were isolated. The frequency of antigen transformants among the TK+ transformants was about 0.5 X 10(-3). The sizes of the HLA, Leu-1, and Leu-2 molecules expressed by the transformants were the same as those of the proteins present on DNA-donor cells.
我们通过使用单克隆抗体进行免疫荧光检测，并利用荧光激活细胞分选仪（FACS）进行筛选，分离出了表达人细胞表面分化抗原的小鼠L细胞稳定转化体。小鼠L细胞（TK-）与人细胞DNA和单纯疱疹病毒胸苷激酶（TK）基因共转化。首先筛选出TK+转化体。用各种针对膜抗原的荧光抗体对TK+群体进行染色，然后使用FACS对阳性细胞进行分选和克隆。分离出了HLA I类抗原、β2-微球蛋白以及T细胞分化抗原Leu-1和Leu-2的转化体。在TK+转化体中，抗原转化体的频率约为0.5×10-3。转化体所表达的HLA、Leu-1和Leu-2分子的大小与DNA供体细胞上存在的蛋白质大小相同。
