Amplification of a gene coding for human T-cell differentiation antigen

Abstract

Using previously isolated mouse L-cell transferees 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)^1,2 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.