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. 2009 Oct 1;119(11):3473–3486. doi: 10.1172/JCI39199

Figure 1. Generation of the Bcl-2–transduced primary CD4+ T cells.

Figure 1

(A) The structure of the lentiviral vector carrying the human Bcl-2 gene (EB-FLV). The U3 region of 3′ LTR is deleted (3′ΔLTR) for self inactivation. The expression of human Bcl-2 is driven by EF1α promoter. Ψ, packaging signal; RRE, rev responsive element; cPPT, central polypurine tract; pEF1α, EF1α promoter. (B) Strategy to generate Bcl-2–transduced primary CD4+ T cells. Primary CD4+ T cells from normal donors were activated and transduced with the Bcl-2–expressing lentiviral vector. Viable cells were isolated after 3 to 4 weeks of culture in the absence of TCR stimulants or cytokines. (C) Intracellular staining for Bcl-2 with FITC-conjugated anti–Bcl-2 antibody in freshly isolated CD4+ T cells and Bcl-2–transduced cells. The Bcl-2–transduced cells were maintained in culture without cytokines for 4 weeks following transduction. Freshly isolated CD4+ T cells stained with FITC-conjugated isotype control antibodies served as a negative control (purple).