Figure 6. In vivo assessment of CCL-βAS3-FB LV transduction of BM CD34⁺ cells.

(A) Engraftment of human cells in NSG mice. BM cells isolated from mice from each transplant group (nos. 1–6) were analyzed by flow cytometry to measure the percentage of human CD45⁺ cells among all CD45⁺ cells in the marrow (human and murine) as a measurement of engraftment. Mock transduced, white triangles; CCL-βAS3-FB transduced, black triangles. BM samples from HD were used in transplants 3, 4, and 6 and from SCD donors in transplants 1, 2, and 5. (B) Immunophenotypic analysis of human cells isolated from NSG mice transplanted with transduced BM CD34⁺ cells. Flow cytometry was used to enumerate the percentage of the human CD45⁺ cells that were positive for the markers of B-lymphoid cells (CD19, white), myeloid progenitors (CD33, light gray), hematopoietic progenitors (CD34, dark gray), and erythroid cells (CD71, black). Mean ± SD are shown of 3 independent experiments. Mock, n = 4; βAS3-FB, n = 8 mice. (C) VC/cell in human cells cultured from NSG mice transplanted with transduced BM CD34⁺ cells. Black circles represent samples from mice transplanted with HD BM, and white squares represent mice transplanted with SCD BM. All the human cells examined from mock-transduced mice were negative for VC analysis by qPCR. (D) HBBAS3 mRNA expression measured by qRT-PCR from cells transduced to different VC/cell. Five independent transductions are shown. HD, black circles (n = 6); SCD, white squares (n = 4).