Loading of antigen on particles as well as the choice of skin as target organ for vaccination were independently described as effective dose-sparing strategies for vaccination. Combining these two strategies, sufficient antigen recognition may be achievable via the transcutaneous route even with minimal-invasive tools. Here, we investigated the skin penetration and cellular uptake of topically administered virus-like particles (VLPs), composed of the HIV-1 precursor protein Pr55gag, as well as the migratory activity of skin antigen-presenting cells (APCs). We compared VLP administration on ex vivo human skin pre-treated with cyanoacrylate tape stripping (CSSS, minimal-invasive) to administration by skin pricking and intradermal injection (invasive). CSSS as well as pricking treatments resulted in penetration of VLPs in the viable skin layers. Electron microscopy confirmed that at least part of VLPs remained intact during the penetration process. Flow cytometry of epidermal, dermal, and HLA-DR+ APCs harvested from culture media of skin explants cultivated at air-liquid interface revealed that a number of cells had taken-up VLPs. Similar results were found between invasive and minimal-invasive VLP application methods. CSSS pre-treatment was associated with significantly increased levels of IL-1α levels in cell culture media as compared to untreated and pricked skin. Our findings provide first evidence for effective cellular uptake of VLPs after dermal application and indicate that even mild physical barrier disruption, as induced by CSSS, provides stimulatory signals that enable the activation of APCs and uptake of large antigenic material.