Figure 4.

β₂ integrins' expression in gene therapy (GT)-treated CD18^HYP mice. Lin⁻ cells from CD18^HYP mice were isolated and transduced with hCD18-LVs. Transduced cells were transplanted into lethally irradiated CD18^HYP mice. As control groups, CD18^HYP mice were transplanted with LV:SFFV.eGFP-transduced CD18^HYP and CD18^WT lin⁻ cells. (A) Percentage of hCD18⁺/mCD11a⁺ peripheral blood lymphocytes (PBLs) at 1 and 3 mpt in ex vivo primary and secondary GT-treated CD18^HYP mice. (B) Mean of mCD11a surface expression levels (mean fluorescence intensity, MFI) in PBLs at the different time points after transplant of transduced cells in primary (black) and secondary (scratched black) CD18^HYP recipient mice and eGFP control animals (dark and light gray). Presented results correspond to six independent experiments. The significance of differences between groups is expressed as ****p < 0.0001 and is always referred to the CD18^HYP + LV:SFFV.eGFP group. (C) Left Y axis shows the percentage of neutrophils, B-cells, and T-cells expressing hCD18 in ex vivo GT-treated primary recipients. Right Y axis shows physiological mCD18 expression pattern. The upper numbers show the ratio of myeloid and lymphoid CD18 expression. (D) Percentage of hCD18⁺ cells in total BMC population of primary and secondary recipients at 4 and 9 months posttransplantation, respectively. (E) mCD11a surface expression levels observed in the BM of transplanted mice. The total number of transplanted animals on each transduction group for any of the hCD18 therapeutic vectors was as follows: in primary transplants, 34 Chim, 17 UCOE, and 27 PGK; in secondary transplants, 7 Chim, 5 UCOE, and 3 PGK. In the case of control animals, 24 were transplanted with CD18^HYP-GFP-transduced cells and 22 with CD18^WT-GFP-transduced cells. The significance of differences between groups is expressed as *p < 0.05 and **p < 0.01. See Supplementary Table S1 for list of antibodies used.