FIG. 3.

gB and gH peptides inhibit pp65 uptake into fibroblast cells. CMV was inoculated onto NHDF cells at an MOI of approximately 0.05 PFU/cell in the presence or absence of gB and gH peptides and allowed to bind and penetrate for 90 min at 37°C. Nonpenetrated virus was inactivated, and the cells were incubated for approximately 18 h at 37°C. Immunofluorescence analysis was performed to detect pp65, and the percentage of pp65-positive cells was scored for each treatment. (A) In addition to inhibiting expression of IE gene products, gB-COIL-30 and gH-COIL inhibited uptake and trafficking of the major tegument protein pp65 to the nucleus of infected cells. Since delivery of pp65 to infected cells is an earlier marker for virus entry than IE gene expression, these data support the hypothesis that the inhibiting peptides act during virus entry and not at the level of transcription or translation of IE gene products. Data are reported in duplicate, and error bars represent the range. (B) Representative immunofluorescence images for mock, no peptide, or gB-COIL-30 treatment showing pp65 in the nucleus of infected fibroblast cells.