Figure 5.

Lentiviral vector–mediated HSC perforin gene transfer restores PRF expression and corrects antigen-specific CTL killing in prf^−/− mice. LV vectors encoding either wild-type human PRF or a mutant human PRF (PRFmut) under the PGK promoter were used to transduce LSK cells from P14.prf^−/− mice, which were transplanted into irradiated prf^−/− recipients. (a) CTL lines derived from these mice were analyzed for GFP expression by flow cytometry, and data is listed as %GFP above the western blot. Perforin expression was analyzed by western blot using Δ G9 anti-perforin Ab. Samples from gene-transduced P14.prf^−/− mice were compared to P14.WT and P14.prf^−/− mice. (b) ⁵¹Cr release from GP33-loaded EL4 target cells co-incubated with CTL from mice reconstituted with PGK.PRF (n = 3) or with PGK.PRFmut (n = 3), P14.WT (n = 5), and P14.prf^−/− mice (n = 3).