Transfer of Anti-Rotavirus Antibodies during Pregnancy and in Milk Following Maternal Vaccination with a Herpes Simplex Virus Type-1 Amplicon Vector

Meier, Anita F.; Suter, Mark; Schraner, Elisabeth M.; Humbel, Bruno M.; Tobler, Kurt; Ackermann, Mathias; Laimbacher, Andrea S.

We conclude that the observed circular structures represent indeed RVLPs formed in the cytoplasm of HSV-1 transduced cells resembling viroplasms found in RV-infected cells. We conclude that the observed circular structures represent indeed RVLPs formed in the cytoplasm of HSV-1 transduced cells resembling viroplasms found in RV-infected cells. The cells were harvested 24 hpt from monolayers by pelleting and fixation with 4% formaldehyde and embedded in LR White. Ultrathin sections were directly collected on grids, prepared for immune fluorescence and images were taken using a fluorescence microscope. Thereafter, the same grids were further processed for electron microscopy. The fluorescent image was opened with the software Maps on the scanning electron microscope (SEM) computer, aligned with the secondary electron image and a tileset of images was recorded using the high-angle annular darkfield detector. (C) Overlay of the fluorescent and the electron microscope image of a stitched tileset of images covering the entire cell of interest. On the fluorescence micrograph, 4′,6-diamidino-2phenylindole (DAPI) was used to stain the nuclei (blue), and RV proteins were stained using the anti-RV serum and the secondary antibody conjugated with Alexa-Fluor 488 (green). The cells were harvested 24 hpt from monolayers by pelleting and fixation with 4% formaldehyde and embedded in LR White. Ultrathin sections were directly collected on grids, prepared for immune fluorescence and images were taken using a fluorescence microscope. Thereafter, the same grids were further processed for electron microscopy. The fluorescent image was opened with the software Maps on the scanning electron microscope (SEM) computer, aligned with the secondary electron image and a tileset of images was recorded using the high-angle annular dark-field detector; (C) Overlay of the fluorescent and the electron microscope image of a stitched tileset of images covering the entire cell of interest. On the fluorescence micrograph, 4 ,6-diamidino-2-phenylindole (DAPI) was used to stain the nuclei (blue), and RV proteins were stained using the anti-RV serum and the secondary antibody conjugated with Alexa-Fluor 488 (green); (D) Enlargement of the area of interest of the stitched tileset showing the highly RV-positive-marked electron-dense region marked in (C). Scale bar: (C,D) 1 µm.