Figure 8. Inhibition of direct polarised epithelial cell infection.

Recombinant EBV carrying a deletion for gp350 (Δgp350) was used to directly infect polarized tonsillar epithelial cells from the basolateral surface. (A) Purified Δgp350 at an MOI 100 was pre-incubated for 1 hour at 37°C with proteins of the extracellular matrix: laminin β-1, fibronectin, vitronectin, heparan sulphate, or expressed on the surface of epithelial cells: ICAM-1 or VCAM-1 as a control. The virus was then incubated for 1 hour with the epithelial cells. Epithelial infection was monitored by flow cytometry for GFP and plotted as % infection (mean of triplicates) relative to the positive control. (B) The Δgp350 virus was pre-incubated with an RGD peptide, a peptide containing the KGD motif specific to gp85 and a peptide containing an RGD motif specific to BMRF2, plus control scrambled peptides in binding buffer for 1 hour at 37°C. The virus was then incubated for 1 hour with the epithelial cells. Epithelial infection was again monitored by flow cytometry for GFP expression and plotted as % infection relative to a positive control without peptides.