Fig. 1: Design and characterization of MVA-VLP-SUDV vaccine.

a MVA-VLP-SUDV construction. Sequences encoding SUDV GP and EBOV VP40 were inserted into the MVA backbone at two distinct sites between essential genes under direction of the modified H5 promoter. b Schematic representation of the mechanisms resulting in high immunogenicity of the MVA-VLP-SUDV construct. c MVA-VLP-SUDV antigen expression. Lysates and supernatant were collected from MVA-VLP-SUDV or parental MVA infected DF1 cells, subjected to Western blotting and probed with antibodies specific for SUDV GP (top) or VP40 (bottom). Expression of SUDV GP and EBOV VP40 was visualized in both the lysate and supernatant of recombinant vaccine infected cells. d VLP formation from MVA-VLP-SUDV infected cells. Thin layer sections of MVA-VLP-SUDV infected DF1 cell monolayers were visualized by TEM and reveal filamentous particle structure budding from the cell surface similar to native SUDV. Immunogold staining is specific for SUDV GP. Representative micrographs are shown with scale bars depicted on the bottom left corner. The scale bar sizes for the three microphotographs, from left to right are: 100 nm, 100 nm, 50 nm.