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. Author manuscript; available in PMC: 2020 Feb 1.
Published in final edited form as: Clin Cancer Res. 2018 Nov 8;25(3):1000–1011. doi: 10.1158/1078-0432.CCR-18-0963

Figure 1. Lentiviral and retroviral vector characterization.

Figure 1.

A. LV-NY-ESO-1 TCR/sr39TK (RRL-MSCV-optNYESO-optsr39TK-WPRE) self-inactivating third-generation lentiviral vector scheme. B. NY-ESO-1 TCR expression in human PBMCs three days after transduction. Percentage of NY-ESO-1 TCR and CD3 expression were measured by flow cytometry with NYESO-1(157–162) dextramer and anti-CD3 antibody in mock-transduced (top panel) and transduced (bottom panel) PBMCs. C. In vitro sr39TK functionality in hCD34+. Mock-transduced or LV-NY-ESO-1 TCR/sr39TK-transduced human CD34+ cells were treated with 0, 0.02, 0.2, 2, 20 or 200μM ganciclovir (GCV) for 48 days. Left, TCR expression (measured by Vβ13 staining) in cells not treated with GCV. Percentage of Vβ13+ (center) and Vβ13- cells (right) in the transduced CD34+ cells after GCV treatment at the indicated concentrations. Vβ13 expression was measure by flow cytometry D. RV-NY-ESO-1 TCR (MSGV1-A2aB-1G4A-LY3H10) gamma-retroviral vector scheme. E. NY-ESO-1 TCR expression in murine T cells from HLA-A2/Kb mice two days after transduction. Surface (top panels) and total (surface + intracellular, bottom panels) TCR expression was measured by Vβ13 TCR beta chain and surface CD3 staining detected by flow cytometry. Abbreviations: Ψ, packaging signal; cPPT, central polypurine tract; LTR, long terminal repeat; MSCV, murine stem cell virus promoter; RRE, Rev response element; SA, splicing acceptor; SD, splicing donor; WPRE, woodchuck hepatitis virus posttranslational response element.