Exploitation of Herpesviral Transactivation Allows Quantitative Reporter Gene-Based Assessment of Virus Entry and Neutralization
Figure 10
Viral transactivation can be neutralized by monoclonal antibodies in MRC-5 and Jurkat cells.
(A) MRC-5 cells were transiently transfected (Lipofectamine 2000 CD) with the pTA-Control plasmid. Cells were infected with HCMV-HB5 (left panel) or HSV-1 strain F (right panel). Replication competent virus (black bars) was compared with UV-inactivated virus (hatched bars). HCMV was incubated for 1 h at 37°C before infection with 25 µg/ml of the HCMV gB-specific monoclonal antibody ITC88 and HSV-1 with the HSV-1 gD-specific monoclonal antibody HD1 before infection. Cells were lysed and luciferase activity was determined. (B) Jurkat cells were transfected with pTA-Control plasmid using the Lonza transfection reagents and protocol. Cells were infected with UV-inactivated HSV-1 strain F (hatched bar), replication competent HSV-1 (black bars) or left uninfected (white bar). Virus was incubated before infection for 1 h at 37°C with indicated dilutions of HSV gD monoclonal antibody HD1. ∼20 h p. i. cells were lysed and the expressed luciferase activity was measured.