Figure 3.

Exogenous IFN-β Reduces LV Transduction Efficiency on Monocytic Cells but Not on HEK293T or Primary T Cells

(A and C) LV encoding GFP was used to infect (A) HEK293T or (C) THP-1 cells in the presence or absence of IFN-β titrations with infection measured by enumerating GFP-positive cells by flow cytometry (mean ± SD, n = 2 biological replicates). TU, transducing units. (B, D, and F) qRT-PCR for the indciated ISGs was performed after 24 h incubation of (B) HEK293T cells, (D) THP-1 cells, or (F) primary T cells stimulated with anti-CD3 and anti-CD28 beads with the indicated IFN-β concentrations (mean ± SD, n = 3 biological replicates). Measurements for each ISG were first normalized to GAPDH and then to mock-treated cells to generate a fold change. (E) Activated T cells were transduced with therapeutic CAR-encoding LV in the presence or absence of increasing amounts of IFN-β and transduction efficiency was assessed at day 5 by flow cytometry with an AffiniPure F(ab’) fragment antibody.