Figure 4
Figure 4. The A2UCOE-EGFP vector gives rise to consistent transgene expression in bone marrow and peripheral blood hematopoietic cells in vivo. HSCs were isolated from the bone marrow of C57BL/6J mice and transduced with the A2UCOE-EGFP, SFFV-EGFP, and CMV-EGFP lentiviral vectors (Figure 1B) at an MOI of 20 to 25 and transplanted into lethally irradiated recipient animals. Total peripheral blood and bone marrow cells were obtained at 3 months after transplantation and analyzed for the presence of EGFP-positive cells by FACS. (A) Percentage of total EGFP-positive cells in bone marrow and peripheral blood from A2UCOE, SFFV, and CMV mice (n = 5 in each group). The horizontal bar shown in each column denotes the mean percentage value of EGFP-positive cells in each group. The P values were determined using the Wilcoxon rank sum test. (B) Expression profiles of a representative mouse that received a transplant in each group. The percentage of total EGFP-positive cells is shown in each plot. Note that the A2UCOE-EGFP vector generates a higher percentage of transgene-expressing cells (panel A) as well as discrete peaks of EGFP-positive cell populations (panel B), suggesting a negation of integration-site position effects leading to more consistent transgene expression compared with that achieved with SFFV and CMV viral promoters. (C) Peripheral blood cells from mice transduced with the A2UCOE-, SFFV-, and CMV-EGFP lentiviral vectors were incubated with antibodies against CD3 (T-cell), CD19 (B-cell), and CD11b (myeloid) lineage markers and scored against EGFP expression by FACS analysis. EGFP expression profile in different cell lineages in a representative mouse that received a transplant, corresponding to panel B (bottom row, peripheral blood), in each lentiviral vector group. (D) Total percentage of EGFP-positive cells in T-, B-, and myeloid cell lineages in peripheral blood from A2UCOE, SFFV, and CMV vector–transduced mice (n = 5 in each group). The horizontal bar shown in each column depicts the mean percentage value of EGFP-positive cells in each cell lineage.

The A2UCOE-EGFP vector gives rise to consistent transgene expression in bone marrow and peripheral blood hematopoietic cells in vivo. HSCs were isolated from the bone marrow of C57BL/6J mice and transduced with the A2UCOE-EGFP, SFFV-EGFP, and CMV-EGFP lentiviral vectors (Figure 1B) at an MOI of 20 to 25 and transplanted into lethally irradiated recipient animals. Total peripheral blood and bone marrow cells were obtained at 3 months after transplantation and analyzed for the presence of EGFP-positive cells by FACS. (A) Percentage of total EGFP-positive cells in bone marrow and peripheral blood from A2UCOE, SFFV, and CMV mice (n = 5 in each group). The horizontal bar shown in each column denotes the mean percentage value of EGFP-positive cells in each group. The P values were determined using the Wilcoxon rank sum test. (B) Expression profiles of a representative mouse that received a transplant in each group. The percentage of total EGFP-positive cells is shown in each plot. Note that the A2UCOE-EGFP vector generates a higher percentage of transgene-expressing cells (panel A) as well as discrete peaks of EGFP-positive cell populations (panel B), suggesting a negation of integration-site position effects leading to more consistent transgene expression compared with that achieved with SFFV and CMV viral promoters. (C) Peripheral blood cells from mice transduced with the A2UCOE-, SFFV-, and CMV-EGFP lentiviral vectors were incubated with antibodies against CD3 (T-cell), CD19 (B-cell), and CD11b (myeloid) lineage markers and scored against EGFP expression by FACS analysis. EGFP expression profile in different cell lineages in a representative mouse that received a transplant, corresponding to panel B (bottom row, peripheral blood), in each lentiviral vector group. (D) Total percentage of EGFP-positive cells in T-, B-, and myeloid cell lineages in peripheral blood from A2UCOE, SFFV, and CMV vector–transduced mice (n = 5 in each group). The horizontal bar shown in each column depicts the mean percentage value of EGFP-positive cells in each cell lineage.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal