a differential role for macropinocytosis in mediating entry of the two forms of vaccinia virus into dendritic cells微分作用对macropinocytosis调解两种牛痘病毒进入树突细胞.pdfVIP

A Differential Role for Macropinocytosis in Mediating Entry of the Two Forms of Vaccinia Virus into Dendritic Cells 1,2¤ 1 1 3 Kerrie J. Sandgren , John Wilkinson , Monica Miranda-Saksena , Gerald M. McInerney , Karen Byth- 1 4 1 Wilson , Phillip J. Robinson , Anthony L. Cunningham * 1 Centre for Virus Research, Westmead Millennium Institute, Sydney, New South Wales, Australia, 2 Faculty of Medicine, University of New South Wales, Sydney, New South Wales, Australia, 3 Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden, 4 Children’s Medical Research Institute, Westmead, Sydney, New South Wales, Australia Abstract Vaccinia virus (VACV) is being developed as a recombinant viral vaccine vector for several key pathogens. Dendritic cells (DCs) are specialised antigen presenting cells that are crucial for the initiation of primary immune responses; however, the mechanisms of uptake of VACV by these cells are unclear. Therefore we examined the binding and entry of both the intracellular mature virus (MV) and extracellular enveloped virus (EV) forms of VACV into vesicular compartments of monocyte-derived DCs. Using a panel of inhibitors, flow cytometry and confocal microscopy we have shown that neither MV nor EV binds to the highly expressed C-type lectin receptors on DCs that are responsible for capturing many other viruses. We also found that both forms of VACV enter DCs via a clathrin-, caveolin-, flotillin- and dynamin-independent pathway that is dependent on actin, intracellular calcium and host-cell cholesterol. Both MV and EV entry