Stable transformation of mouse L cells for human membrane T-cell differentiation antigens, HLA and beta 2-microglobulin: selection by fluorescence-activated cell sorting.
Stable transformation of mouse L cells for human membrane T-cell differentiation antigens, HLA and beta 2-microglobulin: selection by fluorescence-activated cell sorting.
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小鼠 L 细胞稳定转化为人膜 T 细胞分化抗原、HLA 和 β2-微球蛋白：通过荧光激活细胞分选进行选择。
DOI：
10.1073/pnas.80.2.524
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发表时间：
1983
影响因子：
11.1
通讯作者：
Herzenberg,LA
中科院分区：
文献类型：
--
作者：
Kavathas,P;Herzenberg,LA
我们通过使用单克隆抗体的免疫荧光和荧光激活细胞分选仪（FACS）的选择，分离了表达人细胞表面分化抗原的小鼠L细胞的稳定转化体。小鼠 L 细胞 (TK-) 与人类细胞 DNA 和单纯疱疹病毒胸苷激酶 (TK) 基因共转化。首先选择TK+转化体。 TK+群体用针对膜抗原的各种荧光抗体进行染色，并使用FACS对阳性细胞进行分选和克隆。分离了HLA I类抗原、β2-微球蛋白以及T细胞分化抗原Leu-1和Leu-2的转化子。 TK+转化体中抗原转化体的频率约为0.5×10(-3)。转化体表达的 HLA、Leu-1 和 Leu-2 分子的大小与 DNA 供体细胞上存在的蛋白质的大小相同。
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.