| Popis: |
Introduction: Adenovectors are promising vectors for gene delivery to the target cells. During gene therapy, AdV interact with plasma components particularly vitamin K-dependent coagulation factors. In this study, we analyzed the comparison between cell entrance, inflammatory patterns, and innate immune induction which induced by the Adenoventor and Adenovector coated by FVII, FVIII, and FIX on two cell-lines; HepG2, MCF7. Methods: The Adenovector expressing GFP (AdVGFP) was prepared and then loading of AdV by coagulation factors were analyzed by zeta potential measurement. The non-toxic MOI of vector employed alone or in complex with coagulation factors VII, FVIII, and FIX applied on HepG2 and MCF7 cell lines. The transduction rates of complexes were analyzed by fluorescent microscopy and flow cytometry. The expression levels of innate immune genes (PKR, STING, IRF-3 and MX-1) were measured by Real-time PCR. Also the level of IL-6 and IL-1β were evaluated by using ELISA assay. Results: The loading of Adenovector by FVII and FIX decreased the zeta charge of the complex particles and enhanced the entry rate in both HepG2 (FVII/AdV: 38.3% and FIX/AdV: 61.9%) and MCF7 (FVII/AdV: 31.2% and FIX/AdV: 36.6%). The expression of IL-6 cytokine enhanced when AdV exposed to FVII (P value: 0.005) in MCF7 and also FVII (P value: 0.01) and FIX (P value: 0.009) in HepG2. Adenovector coated by FVII and FIX could significantly alter the expression pattern of innate immune genes. Conclusion: The findings are highlighted the role of FVII and in particular FIX in facilitating the entry of vector into the cells; also they are enhanced the inflammation and innate immune responses. Interestingly, FVIII had no effect or even adverse effect on entry, inflammation, and innate immune induction. |
