Induction of cell surface insulin antigenicity in fibroblasts transfected with islet genomic DNA.
Ltk- cells were transfected with total genomic DNA obtained from rat islets or from insulinoma cells mixed with DNA tagged with fluorescent dye. Fluorescence-activated cell sorter (FACS) was used for initial selection of successful transfectants, monitoring fluorescent DNA incorporated into the cell. For subsequent selections the cells were treated with anti-insulin antiserum labeled with fluorescent dye and selected by FACS. Beta-cell DNA, but not DNA from murine leukemic cells, induced the appearance of cell surface insulin antigenicity in fibroblasts. This phenotypic expression was transient. Together with our previous demonstration of the induction of insulin secretion in Ltk- cells [(1986) Biochem. Biophys. Res. Commun. 136, 638-644], these results indicate that beta-cell-specific characteristics can be transfected to non-endocrine cells by genomic DNA transfection.
D000818:Animals; D000941:Antigens / Q000032:analysis*; D002469:Cell Separation; D004247:DNA / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics; D005434:Flow Cytometry; D007328:Insulin / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D000078790:Insulin Secretion; D007340:Insulinoma / Q000235:genetics; D007515:Islets of Langerhans / Q000032:analysis; D007739:L Cells / Q000032:analysis / Q000276:immunology*; D007942:Leukemia, Experimental; D008297:Male; D010190:Pancreatic Neoplasms / Q000235:genetics; D051381:Rats; D014162:Transfection
D000941:Antigens; D004273:DNA, Neoplasm; D007328:Insulin; D004247:DNA