Vaccine vectors based on low seroprevalent adenovirus type 26 (HAdV26) are listed as intervention in 28 clinical trials, among which several in phase III. While HAdV26 immunogenicity in vivo is rather well described, basic biology of this virus is still insufficiently understood. Recently we reported αvβ3 integrin as a receptor required for HAdV26 successful infection of epithelial cells, however studies regarding HAdV26 endocytosis and innate immunity induced by HAdV26 infection of epithelial cells are missing. Based on our preliminary data we hypothesize that altered HAdV26 endocytosis due to binding αvβ3 integrin or modified clathrin, could lead to induction of different host innate immune response, hence influence performance of HAdV26-based vector. Therefore, the goal of AdenoIN is to investigate role of clathrin in HAdV26 transduction and address clear gap in understanding the innate immune response induced by HAdV26 infection of epithelial cells in the context of using αvβ3 integrin as a receptor and/or altered virus endocytosis. Final output of AdenoIN will be comprehensive analysis of αvβ3 integrin and clathrin mediated HAdV26 endocytosis and subsequent innate immune signaling in epithelial cells. Data and knowledge obtained within AdenoIN will be valuable for further improvement of HAdV26-based vectors.