engleski

Measuring innate immune response in epithelial cells after adenovirus infection

Adenovirus vectors based on a serotype 5 (AdV5) are the most commonly investigated adenoviral vectors for gene therapy and vaccination. Limitations for using AdV5 as a vector are activation of the host innate immune responses as well as preexisting immunity to AdV5 wild type. The aim of this study was to investigate the differences in the activation of innate immune response after infection with AdV5 and low seroprevalent serotypes of adenovirus: AdV35, and AdV26, which may be used to overcome the preexisting immunity to AdV5. After infecting SK-OV-3 and A549 cell lines with replication deficient AdV5, AdV26 and AdV35, we measured the levels of mRNA and corresponding proteins for molecules known to be involved in innate immune response using qPCR, western blot and ELISA. We showed that in contrast to AdV5, infection with AdV26 and AdV35 increased expression of genes known to be involved in initiation of inflammation, namely IL-6, IL-8, IFN-α1, TNF-α and IL-1β. We also observed differences in adaptor molecules situated downstream of pattern recognition receptors and those involved in inflammasome formation. We hypothesize that there are differences in role of innate immune response in AdV26 and AdV35 infection which might be used for construction of vectors with better performances for DNA transfer and vaccination purposes.

adenovirus ; innate immune response ; gene transfer and vaccination

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