Immortalization of human primary B lymphocytes in vitro with DNA.

AUTOR(ES)

Kempkes, B

RESUMO

Epstein-Barr virus (EBV) is a human DNA tumor virus that efficiently immortalizes human primary B lymphocytes in vitro. Although viral genes that are expressed in latently infected B lymphocytes have been shown to function in cellular growth control, their detailed genetic analysis has been cumbersome for two reasons. The viral genome is too large to permit genetic engineering and human primary B lymphocytes, the only targets for infection by EBV in vitro, are both intractable in culture and recalcitrant to DNA transfection. To overcome these obstacles, we have assembled all the essential genes of EBV on a single recombinant vector molecule in Escherichia coli. We show here that this mini-EBV plasmid can yield immortalized B cells upon transfer of its naked DNA into human primary B lymphocytes. Established cell lines carry recombinant vector DNA and cannot support virus production. Because this DNA can be easily manipulated in E. coli, mutant mini-EBVs as well as foreign genes can now be introduced and studied successfully in recipient B lymphocytes from any human donors. These mini-EBVs therefore are potentially useful for human gene therapy.

Documentos Relacionados

• Immortalization of human B lymphocytes by a plasmid containing 71 kilobase pairs of Epstein-Barr virus DNA.
• Short DNA sequences from the cytoplasm of mouse tumor cells induce immortalization of human lymphocytes in vitro.
• Evidence for increased in vitro recombination with insertion of human hepatitis B virus DNA.
• Immortalization of human lymphocytes by transfection with DNA from mouse L929 cytoplasts.
• Role of human T-cell leukemia virus type 1 X region proteins in immortalization of primary human lymphocytes in culture.