Clonal propagation of Epstein-Barr virus (EBV) recombinants in EBV-negative Akata cells

N Shimizu, H Yoshiyama, K Takada
1996 Journal of Virology  
We lack a host cell supporting an efficient lytic replication of Epstein-Barr virus (EBV). Recently, we isolated EBV-negative cell clones from the Akata cell line (referred as Akata ؊ [N. Shimizu, A. Tanabe-Tochikura, Y. Kuroiwa, and K. Takada, J. Virol. 68:6069-6073, 1994]). Since the parental Akata line is one of the highest EBV producers, we examined whether Akata ؊ cells had become a good host for EBV propagation. The parental Akata cells have about 20 copies of EBV plasmid per cell. A drug
more » ... id per cell. A drug resistance gene was inserted into one of them by homologous recombination. The resultant virus preparation, a mixture of wild-type and recombinant EBV, was used to infect Akata ؊ cells. After incubation in the selective medium, drug-resistant Akata ؊ cell clones were isolated and proved to be infected with recombinant EBV only. By treatment of the cells with antiimmunoglobulin antibodies, a large amount of recombinant EBV (i.e., more than 10 g/1-liter culture) was produced. In contrast, three other B-lymphoma lines, BJAB, Ramos, and Louckes, were nonpermissive for virus replication. These results indicate that Akata ؊ cells are suitable for propagation of recombinant EBV clonally, which becomes a powerful tool for determining EBV genetics and which makes it possible to use EBV as a vector for gene therapy. on May 10, 2020 by guest Downloaded from 7262 NOTES J. VIROL. on May 10, 2020 by guest Downloaded from FIG. 4. Schematic representation of the Akata cell system, which allows propagation of EBV recombinants clonally and in large quantities. neo r , Neo r gene.
doi:10.1128/jvi.70.10.7260-7263.1996 fatcat:h6sz3b63pzcefoe2tidijvznfa