Transcriptional Targeting of Mature Dendritic Cells with Adenoviral Vectors via a Modular Promoter System for Antigen Expression and Functional Manipulation

Figure 2

Generation and evaluation of two different Hsp70B heat shock response elements. (a) Plasmid vectors containing Hsp70B_−29/−489 and Hsp70B_−29/−242 were generated by digesting the human hsp70B gene 5′-region with HindIII/BamHI or HindIII/SmaI, respectively, followed by ligation into the pGL3Basic vector. (b) HeLa cells were transfected with 0.5 μg of plasmid DNA of pGL3-SV40-promoter vector (“SV40”), promoterless pGL3-Basic vector (“Basic”), pHsp70B_−29/−489, or pHsp70B_−29/−242. The next day, cells were either left untreated (“Mock”) or transduced with adenoviruses Ad5Luc1 or Ad5mHSF1 at 300 TCID₅₀/cell. Twenty-four hours later, cells were harvested and analyzed by luciferase reporter assays. Results are shown as fold induction relative to the individual pGL3-SV40-promoter control containing no virus, Ad5Luc1 or Ad5mHSF1. Data are mean ± SEM of three independent experiments. , n.s.: not significant (); bars without annotation are not significant (); two-way ANOVA with Bonferroni’s Multiple Comparison post hoc test.

(a)

(b)