Isolation of Epstein-Barr virus-infected clones of the human T-cell line MT-2: use of recombinant viruses with a positive selection marker

Abstract

Certain classes of human T-cell lymphomas have been shown to be persistently infected with Epstein-Barr virus (EBV). To achieve an experimental system of persistent EBV infection in T cells, we used EBV recombinants with a positive selection marker. Infection of the human T-cell line MT-2 with EBV recombinants that had a hygromycin resistance gene and subsequent selection with this drug permitted isolation and long-term maintenance of EBV-infected MT-2 clones. For each clone, essentially 100% of cells were positive for EBV nuclear antigen. These MT-2 clones harbor monoclonal episomes of EBV DNA and stably express two EBV latent proteins, EBV nuclear antigen 1 and latent membrane protein 1. The growth of these EBV-infected MT-2 clones was slower than that of uninfected clones, suggesting that EBV affects regulation of T-cell proliferation. EBV recombinants with a positive selection marker will be a useful tool in establishing experimental systems of persistent EBV infection in certain non-B-cell lineages.