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. 2022 Apr 22;13:852830. doi: 10.3389/fimmu.2022.852830

Figure 1.

Figure 1

Ex vivo lentiviral mediated transduction delivers ADA2 protein expression and enzyme activity with no detrimental effect to the proliferative potential and colony forming capacity of CD34+haematopoietic stem progenitor cells (HSPC). (A) Relative ADA2 mRNA expression was examined in FACS sorted lymphocytes, monocytes, NK cells and neutrophils derived from both healthy controls (n = 2) and patients with DADA2 (n = 2). Both monocytes and lymphocytes were confirmed to express ADA2. (B) Lentivirus construct design containing codon optimised ADA2 c-DNA employing an EFS promoter and tagged to GFP. (C) ADA2 enzymatic activity was increased in macrophages derived from EFS-ADA2-GFP transduced healthy control CD34+HSPC compared to EFS-GFP alone treated cells (n = 3). (D, E) EFS-ADA2-GFP transduction of healthy control CD34+HSPC had no impact on cell proliferation and colony forming capacity across all myeloid and erythroid lineages (n=3). DADA2, deficiency of adenosine deaminase type 2; 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; NK, natural killer; UT, untransduced. WPRE, Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element.