Transcriptional Targeting of Mature Dendritic Cells with Adenoviral Vectors via a Modular Promoter System for Antigen Expression and Functional Manipulation
The constitutively active mHSF1 induces specific transgene expression in HeLa cells and human DCs. (a, b) HeLa cells were transfected with 0.7 μg DNA of plasmid vectors expressing MelanA/MART-1 (“MelA”), BclxL or/and IL-12 under the control of the Hsp70B′−29/−242 heat shock response element as indicated. The next day, cells were either left untreated (“Mock”) or transduced with adenoviruses Ad5Luc1 or Ad5mHSF1 at 300 TCID50/cell. (c, d) Human immature DCs were cotransduced in the presence of the maturation cytokine cocktail with adenoviruses as indicated at a total amount of 500 TCID50/cell. (a–d) Twenty-four hours later, cell lysates were analyzed for BclxL and MelanA expression by Western Blot (a, c) and cell culture supernatants for content of IL-12p40 and IL-12p70 by ELISA (b, d). (a and c) Showing one representative experiment out of three; (b and d) data are mean ± SEM of three independent experiments with different donors. , , and n.s.: not significant (); bars without annotation are not significant (); one-way ANOVA with Bonferroni’s Multiple Comparison post hoc test.
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