FIG. 4.

Comparison of the uptake of AAVLP-OVA and wtAAVLP by different cell types by immunofluorescence microscopy. Cells grown on glass cover slips were incubated for 2 h with AAVLP-OVA, wtAAVLP, or FITC-dextran, followed by incubation for 18 h after removal of the AAVLPs. The cells were then fixed and processed for immunofluorescence microscopy. (A) AAVLP uptake into HeLa epithelial cells and RAW264.7 macrophages. The localization of the VLPs in close vicinity to the nucleus in both cell lines after 18 h of chase incubation is indicative of cellular uptake of the virus particles (compared to virus localization at 2 h after VLP addition). (B) Uptake of AAVLPs or FITC-dextran into mouse bone marrow–derived DC. In the last row confocal images of anti-CD11c-labeled cells are shown (one central layer), while the other images are non-confocal and the nuclei are labeled. The presented images are representative for the results of at least two independent experiments. The slight red staining seen in one cell in the anti-CD11c-labeled AAVLP-OVA-treated DC is most probably background staining, as a similar staining was also seen in some untreated DC. DC, dendritic cell.