Affiliations: SmithKline Beecham Pharmaceuticals, Department of Cellular Biochemistry and Immunology, King of Prussia, PA, USA
Note: [] Address reprint requests to Dr. Zdenka L. Jonak, Department of Cellular Biochemistry and Immunology, 709 Swedeland Road (UW2109), King of Prussia, PA 19406-2799, USA.
Note: [] Katherine Muirhead's present address is Zynaxis, 371 Phoenixville Pike, Malvern, PA 19355, USA.
Abstract: Electroporation was used to deliver genomic DNA from a lymphoid tumor to activated/stimulated human peripheral blood lymphocytes to create immortalized lymphoid cell lines. Activation of the recipient lymphocytes was essential for efficient immortalization. A panel of human B cell transfectant clones, each phenotypically representing specific stages of differentiation, resulted from the transfection. Monoclonal antibody production was measured, and the level produced depended on the phenotype of the cells, with the more mature B cell transfectants secreting up to 10 μg/mL of immunoglobulin. The transfectants were stable with respect to their morphological appearance, growth rate, and antibody production. Chromosome analysis indicated that the transfectants displayed a normal karyotype, devoid of abnormalities. We have shown that electroporation is an effective method of immortalizing human lymphocytes at different stages of differentiation. The transfectants provide a panel of cells that can readily be studied with respect to their phenotypic/karyotypic stability, regulation, and production of immunoglobulin, lymphokines, and growth factors. These data demonstrate the feasibility of generating immortalized human B cells to provide an important resource for the study of B cell differentiation and immortalization.
Keywords: immortalization, novel cell lines, human monoclonal antibodies, in vitro immunization, oncogenic DNA, electroporation
