Interference of HSV cell-to-cell spread by expression of gEΔCT and gI in cells

HSVAd vectoraSize of HSV plaquesbNo. of plaquescYield of HSVd
Wild typeNo Ad100100320
AdTet-trans94.4 ± 2.896ND
AdtetgI94.6 ± 7.8116ND
AdgE(E1−)/gI96.1 ± 4.496350
AdgEΔCT/gI35.5 ± 3.992280
AdgD52.7 ± 5.611ND
AdgEΔCT/gI and AdgD30.1 ± 3.49ND
F-gEβNo Ad29.7 ± 0.6100ND
AdgEΔCT/gI30.2 ± 0.495ND
  • a HaCaT cells were infected with various Ad vectors or combinations of vectors with 300 PFU/cell for each of the single infections with AdTet-trans, AdtetgI, and AdgD; 150 PFU of each virus per cell for the dual infections, AdgE(E1−)/gI and AdgEΔCT/gI; and 100, 100, and 200 PFU/cell, respectively, for the AdgEΔCT-AdtetgI-AdgD infections. After 12 h, the cells were infected with wild-type HSV-1 strain F or the gE-null mutant, F-gEβ, with ∼100 PFU/dish for 48 to 60 h.

  • b The cell monolayers were fixed and stained with polyclonal anti-HSV antibodies, secondary peroxidase-conjugated antibodies, and peroxidase substrate. The diameter of ∼20 plaques was measured, and standard deviations are shown.

  • c The numbers of plaques produced on cells not infected with Ad vectors were arbitarily set at 100, and plaques formed on other monolayers were compared to this.

  • d Cells infected with Ad vectors were infected with HSV with 10 PFU/cell for 18 to 20 h, then the cells and media were harvested and sonicated, and infectious HSV titers were determined on Vero cell monolayers. Yield of HSV indicates PFU per cell. ND, not determined.