Successful reconstitution of T cells, B cells, and NK cells in γ_c-deficient mice by a gene therapy approach. (A) Increased thymocytes after reconstitution of γ_c-deficient mice with γ_c-deficient bone marrow that was transduced with human γ_c. Note that the ordinate is a log scale. (B and C) Increased splenic T cells (B) with normalization of the CD4:CD8 ratio (C) in γ_c-deficient mice with γ_c-deficient bone marrow that was transduced with human γ_c; (B) also indicates correction of the B-cell defect in splenocytes. (D) Increased numbers of splenic TCR⁻NK1.1⁺ cells after reconstitution of γ_c-deficient mice with γ_c-deficient bone marrow that was transduced with human γ_c. Note that the TCR⁺NK1.1⁻ cells are partially diagonally shifted to the right (albeit still to the left of the NK1.1⁺ cells that are boxed in each dot plot); this is an artifact that resulted from the use of 4-color flow cytometry. (E) Increased B cells in bone marrow after reconstitution of γ_c-deficient mice with γ_c-deficient bone marrow that was transduced with human γ_c. In (B through E), we have selected representative dot plots for each of the cell types analyzed from our final 3 different gene therapy experiments in which all analyses were performed. Our initial experiments provided consistent information, but not all assays were included in those experiments. In the final 3 experiments, splenic cellularities (mean ± standard error of mean [SEM] × 10⁻⁶) were as follows: wild-type mice (87.0 ± 3.0, top dot plots), γ_c-deficient mice (49.0 ± 5.5, second dot plots), γ_c-deficient mice, which received wild-type bone marrow (77.8 ± 19.9, third dot plots), γ_c-deficient mice, which received human γ_c-transduced bone marrow (82.8 ± 15.6, fourth dot plots), and γ_c-deficient mice, which received mock-transduced bone marrow (20.8 ± 3.9, bottom dot plots). The data in (A) were from the final 2 experiments and those in (B) through (E) were representative of the final 3 experiments. The number of mice analyzed in each experiment was as follows: experiment 1 (mice analyzed at 63 days after bone marrow transplantation): 1 wild-type mouse, 2 γ_c KO mice (no treatment), 2 γ_c KO mice (WT-BMT), 3 γ_c KO mice (hγ_c-transduced), and 1 γ_c KO mouse (mock-transduced). Experiment 2 (mice analyzed at 62 days after bone marrow transplantation): 2 wild-type mice, 2 γ_c KO mice (no treatment), 1 γ_c KO mouse (WT-BMT), 2 γ_c KO mice (hγ_c-transduced), and 1 γ_c KO mouse (mock-transduced). Experiment 3 (mice analyzed at 52 days after bone marrow transplantation): 1 wild-type mouse, 2 γ_c KO mice (no treatment), 1 γ_c KO mouse (WT-BMT), 3 γ_c KO mice (hγ_c-transduced), and 2 γ_c KO mice (mock-transduced). Experiment 3 was not included in (A) because thymocyte numbers were not counted.