Figure 1.

Vpx-Containing Lentiviral Vectors Expressing CD40L Efficiently Transduce and Activate HSC-DCs

(A) Diagrams of the ∼5.2-kb lentiviral vectors expressing mutant CD40L (mtCD40L), CD40L-M1, or CD40L-SL9 and the Vpx motif chimeric Gag/Pol and SIVmac Vpx expression vectors used to produce Vpx-containing virions are shown. 293T cells cotransfected with the vectors and expression vectors for VSV-G and HIV-1 Rev (not shown) yield Vpx-containing lentiviral vector stocks. (B) HSC-DCs were transduced with the Vpx-containing lentiviral vectors and after 72 h stained with anti-CD40L-PE and analyzed by flow cytometry to determine the percentage of CD40L⁺ cells. The results shown are representative of three different donor HSC-DCs. (C) The expression of HLA-DR, CD83, and ICAM-1 on the transduced HSC-DCs was quantified by flow cytometry. Pooled results from five donors are shown. Data represent mean ± SD. *p < 0.05, **p < 0.01 by unpaired two-tailed t test with Welch’s correction. (D) IL-6, IL-12p70, and TNF-α in the transduced HSC-DC supernatant were quantified by cytokine bead array. Data represent triplicate samples from one donor with mean ± SD. nd, not detected. Additional donors are shown in Figure S2C.