Kavathas P, Herzenberg L A
Nature. 1983;306(5941):385-7. doi: 10.1038/306385a0.
Using previously isolated mouse L-cell transferents for the human T-cell differentiation antigen Leu-2, we now report the first example of spontaneous gene amplification for membrane antigens. The Leu-2 (or T8) antigen is normally expressed on T lymphocytes that have cytotoxic or suppressor functions. Cells of a Leu-2 transfected clone were stained with fluorescein-tagged monoclonal anti-Leu-2, and the brightest 0.1-0.3% of cells were viably separated using a fluorescence activated cell sorter (FACS). After growth of these selected cells, sorting and regrowth was repeated six times, resulting in a population of cells that, compared with the starting population, stains 40 times brighter for Leu-2 and whose DNA transforms 20 times more efficiently for Leu-2. In addition, these cells have 10- to 50-fold amplified human DNA sequences and numerous double minute chromosome fragments, a common indicator of gene amplication in mouse cells.
利用先前分离得到的携带人类T细胞分化抗原Leu-2的小鼠L细胞转染体，我们现在报道膜抗原自发基因扩增的首个实例。Leu-2（或T8）抗原通常在具有细胞毒性或抑制功能的T淋巴细胞上表达。用荧光素标记的抗Leu-2单克隆抗体对Leu-2转染克隆的细胞进行染色，然后使用荧光激活细胞分选仪（FACS）将最亮的0.1 - 0.3%的细胞进行活细胞分选。这些分选后的细胞生长后，重复分选和再生长过程六次，最终得到一群细胞，与起始细胞群体相比，这群细胞对Leu-2的染色亮度提高了40倍，其DNA对Leu-2的转化效率提高了20倍。此外，这些细胞具有10至50倍扩增的人类DNA序列以及大量双微小染色体片段，这是小鼠细胞中基因扩增的常见指标。
