Methods for manufacturing “off-the-shelf” EBV-specific cytotoxic T cells. There are several methods for manufacturing off-the-shelf EBV-specific CTLs. This figure shows 2 common approaches. The cells are collected from a healthy donor. Peripheral blood mononuclear cells (PBMCs) are selected and stimulated using 1 of several methods. Method 1 uses the nucleofection of plasmid DNA into dendritic cells. These dendritic cells are used to present antigen to cytotoxic T cells. This is followed by the selection of cytotoxic T cells with high levels of interferon gamma (IFN-γ) secretion. This method takes 2 to 3 weeks in total. Method 2 uses overlapping peptide mixes or pepmixes of EBV nuclear antigen to stimulate PBMCs. The T cells are, then, selected and expanded in a closed system. This method takes ∼10 days to manufacture the EBV CTLs. EBV CTLs are banked and can be infused into HLA-matched recipients.