Figure 4.

LentiBOOST (LB) increases transduction efficiency of CB CD8⁺-transduced T cells and maintains proliferation in vitro

(A) Transduction efficiency, presented as percentage of GFP⁺ cells, is improved by the use of LB together with LV at MOI 10 (left) and MOI 1 (right). (B) Protein expression, calculated as MFI of EGFP signal, is increased by the use of LB together with both LV at MOI 10 (left) and MOI 1 (right). (C) qPCR analysis of the VCN in CB CD8⁺ T cells transduced with an MOI 10 in the absence (black) or presence (black stripes) of LB or transduced with an MOI 1 in the absence (gray) or presence (gray stripes) of LB. (D) Representative flow cytometry plot of proliferation analysis using CellTrace Violet (CTV) on CB CD8⁺ T cells transduced with the STND method or with STND+LB. (E) Proliferation rate (as CTV-MFI of day 0/day 4) in CB CD8⁺ T cells untransduced and transduced in the presence or absence of LB. (F) Proliferation indexes (calculated using proliferation model of FlowJo v.10) define proliferation, division, and expansion rate of CB CD8⁺ T cells transduced in the presence of LB (black stripes) compared to STND protocol (black). (G) Cell viability, respectively, at day 0 and day 4 of proliferation assay in CB CD8⁺ T cells transduced in the presence of LB (black stripes) compared to STND protocol (black). The data correspond to ≥3 independent experiments and are shown as average ± standard deviation; ∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001.