Figure 1
Cutaneous adenoviral gene delivery. (A) Baseline expression of cytokines and potentially in cytoplasmatic nucleic acid recognition involved receptors or adapter molecules in HaCaT cells and primary human keratinocytes (HKC). Data was generated via qRT-PCR and is displayed as mRNA concentration in ng/g 18S rRNA (* = p < 0,05; ** = p < 0,005). (B) Comparative efficiency of adenoviral vectors in HaCaT cells amd HKC. Quantitative analysis of GFP-positive cells 48 hours after transfection or transduction of HaCaT cells and HKC with 5 μg adenoviral DNA (AdDNA[5μg]) or 1.1 × 109 IU (AdV[0,5μg]) and 1.1 × 1010 IU (AdV[5μg]) adenoviral vectors (* = p < 0,05; ** = p < 0,005). (C) Comparative GFP expression in HaCaT cells and HKC. The specific transcript was determined 48 h after transfection with 5 μg/ml medium of isolated adenoviral DNA (AdDNA[5μg]) and compared to transduction with 1.1 × 109 IU (AdV[0,5μg]) and 1.1 × 1010 IU (AdV[5μg]) of a GFP-encoding adenoviral vector (* = p < 0,05; ** = p < 0,005). (D) Comparative type-I-interferon expression in HaCaT cells and HKC. The specific transcript was determined 6 h after transfection with 5 μg/ml medium of isolated adenoviral DNA (AdDNA[5μg]) and compared to transduction with 1.1 × 109 IU (AdV[0,5μg]) and 1.1 × 1010 IU (AdV[5μg]) of a GFP-encoding adenoviral vector (* = p < 0,05; ** = p < 0,005). (E) Comparative cytokine expression after transfection of HaCaT cells and HKC with 5 μg/ml isolated adenoviral DNA for 15 h (HaCaT) or 6 h (HKC). These time points were determined (maximum expression) by a time course of cytokine expression in hacats in a previous study by Steinstraesser et al. (Steinstraesser et al. 2011). Study groups included n = 18 (type-I-IFN) or n = 6 (cytokines) samples. Data was normalized to a vehicle control (* = p < 0,05; ** = p < 0,005).