Due to their ability to trigger strong immune responses, adenoviruses (HAdVs) in general and the serotype5 (HAdV5) in particular are amongst the most popular viral vectors in research and clinical application. However, efficient transduction using HAdV5 is predominantly achieved in coxsackie and adenovirus receptor (CAR)-positive human cells. In the present study, we used the transduction enhancer LentiBOOST® comprising the polycationic Polybrene to overcome these limitations. Using

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... Polybrene, we yielded transduction rates higher than 50% in murine bone marrow derived dendritic cells (BMDCs), while maintaining their cytokine expression profile and their capability to induce T-cell proliferation. In human dendritic cells (DCs), we increased the transduction rate from 22% in immature (i)DCs or 43% in mature (m)DCs to more than 80%, without inducing cytotoxicity. While expression of specific maturation markers was slightly upregulated using LentiBOOST®/ Polybrene on iDCs, no effect on mDC phenotype or function was observed. Moreover, we achieved efficient HAdV5 transduction also in human monocytes and were able to subsequently differentiate them into proper iDCs and functional mDCs. In summary, we introduce LentiBOOST® comprising Polybrene as a highly potent adenoviral transduction agent for new in vitro applications in a set of different immune cells in both mice and humans.