Skip to main content
. Author manuscript; available in PMC: 2009 Sep 1.
Published in final edited form as: Best Pract Res Clin Haematol. 2008 Sep;21(3):405–420. doi: 10.1016/j.beha.2008.06.002

Figure 1. Generation of LMP2-specific cytotoxic T lymphocytes (CTLs).

Figure 1

(a) Lymphoblastoid cell line (LCL) generation and transduction with Ad5F35LMP2 vector. An LCL is generated by infecting patient peripheral blood mononuclear cells (PBMCs) with a laboratory strain of Epstein-Barr virus (EBV B95-8). The resultant LCL is then transduced with the Ad5F35LMP2 vector, cultured for 2 days, and then either frozen for future CTL stimulations or used fresh to stimulate and expand LMP-specific CTLs. (b) Dendritic cell (DC) generation and transduction with Ad5F35LMP2 vector. DCs are generated from patient PBMCs (obtained from a fresh blood sample) in the presence of GM-CSF and interleukin (IL)-4 for 5 days. These immature DCs are then transduced with an Ad5F35LMP2 vector and matured in the presence of GM-CSF, tumour necrosis factor-α, IL-4 and PGE1. After 2 days, the Ad5F35LMP2-transduced DCs are irradiated and used to initiate the LMP2-CTL. (c) Generation of LMP2-specific CTLs. Patient PBMCs are co-cultured with autologous, irradiated transduced DCs at a ratio of 20 PBMCs to 1 DC. Cultures are then stimulated on day 10 with irradiated Ad5F35LMP2-transduced LCL at a ratio of 4:1. IL-2 is first added on day 13 and then used twice weekly thereafter. Weekly stimulations with transduced LCLs in the presence of IL-2 are continued until sufficient numbers of LMP2-specific CTLs are generated.