Rapidly-Generated EBV-Specific T Cells (Ebvst-Cells) to Treat Type 2 Latency Lymphoma

Up to 30 % of Hodgkin and non-Hodgkin lymphomas carry the Epstein Barr virus (EBV) genome and express the viral latency type 2 proteins EBNA-1, LMP-1, LMP-2 and BARF-1 in a pattern termed Type 2 latency. We have previously shown that Epstein-Barr virus-specific T-cells (EBVSTs) can be expanded from the peripheral blood of lymphoma patients by stimulation with dendritic cells and autologous EBV-transformed lymphoblastoid cell lines (LCLs) modified with an adenoviral vector encoding LMP1 and LMP2. These EBVSTs induce complete clinical responses in over 50% of patients with active disease (Bollard et al J Clin Oncol 2014). Since the requirement for an LCL as an antigen-presenting cell added 3-4 months to the manufacturing time, we shortened and simplified the process. We replaced the LCL with autologous, activated T-cells together with HLA-negative K562 costimulatory cells as a source of antigen-presenting cell and replaced live virus (EBV) and adenoviral-vector components as a source of antigen with overlapping peptide libraries (pepmixes) spanning the all 4 Type 2 latency antigens (EBNA1, LMP1, LMP2 and BARF1). We also added IL4 and IL7, cytokines that increased the repertoire and expansion of EBVSTs as previously demonstrated by Gerdemann et al. (Mol Ther 2012) These changes reduced the manufacturing time to 3-4 weeks and increased the number of eligible patients, since patients with a short life expectancy were previously excluded and LCLs could not be generated from patients who had received the B-cell depleting antibody rituximab.