Adoptive Immunotherapy against Posttransplant Lymphoprolifrative Disorders

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There is increasing interest in the use of immunotherapy with antigen-specific cytotoxic T cells (CTL) for the treatment of Epstein-Barr virus (EBV)-associated postransplant lymphoproliferative disorders (PTLD). The present study is aimed to develop a quantitative RT-PCR test for monitoring EBV reactivation in the peripheral blood and to test the safety, persistence and anti-tumor activity of in vitro generated CTL. EBV infection was quantified by realtime PCR assay using Lightcycler. The cellular DNA was purified by EZNA blood kit. The primer pair was set at the latent membrane protein 2A region. This method could detect 1 copy of EBV genome per one million peripheral mononuclear cells. The total time required was one hour. The EBV-specific CTL was induced by cocultivation of peripheral blood mononuclear cells with autologous EBV-immortalized lymphoblastoid cells (LCL) that were killed by γ ray irradiation. They were incubated in the presence of IL2 for 2 to 4 weeks and subjected to cytotoxicity test. The result indicated that the CTL was specific for autologous LCL.