Figure 2.

Ex vivo lentiviral mediated gene transfer restores ADA2 protein expression, enzyme activity and ameliorated the proliferative potential and colony forming capacity of CD34+cells from a patient with DADA2. (A, B). EFS-ADA2-GFP transduction of CD34+HSPC derived from a patient with DADA2 with bone marrow aplasia (genotype ADA2 p.G47W/p.G47W) improved the ability of these cells to proliferate and their colony forming capacity across all myeloid and erythroid lineages. (C, D). In addition, there was recovery of cellular ADA2 protein expression and enzyme activity in macrophages derived from EFS-ADA2-GFP transduced patient CD34+HSPC compared to EFS-GFP alone treated cells. (E) There was also reduction in the levels of proinflammatory cytokines (TNF-α, IL-6 and IFN-γ) released by macrophages derived from EFS-GFP-ADA2 transduced patient CD34+HSPC compared to EFS-GFP alone treated cells. Blots were performed in cell lysates. DADA2, deficiency of adenosine deaminase type2; HSPC, haematopoietic stem cells; EFS, elongation factor 1α short; GFP, green fluorescent protein; ADA2, adenosine deaminase 2; GEMM, granulocyte, erythroid, macrophage, megakaryocyte; GM, granulocyte–macrophage; BFU-E, burst-forming units erythroid; UT, untransduced; TNF, tumour necrosis factor; IL, interleukin; IFN, interferon.