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.

HepG2 cells were transduced (MOI 5) with the HSV-1 amplicon vector sWa[VP2/6/7]. After 24 h, the cells were scraped into the medium and resuspended in 2.5% glutaraldehyde (GA), centrifuged for 20 min at 4000 g and the pellet was embedded in Epon according to a standard protocol previously described [50] . Briefly, the cell pellet was fixed with 2.5% GA and postfixed with 1% osmium tetroxide. Before embedding in epoxy resin (Epon), the cell pellets were dehydrated through series of solvents. Finally, ultrathin sections were stained with uranyl acetate and lead citrate and coated with carbon. Specimens were analyzed in a transmission electron microscope (CM12, Philips) equipped with a CCD camera (Orius SC1000W, Gatan) at an acceleration voltage of 100 keV.