Construction of a Vero Cell Line Expressing Human ICAM1 for the Development of Rhinovirus Vaccines

Figures

• 

  Figure 1. Basal surface expression of ICAM1, CD155 (the poliovirus receptor) and CAR on Vero cells as determined by staining with PE-labelled antibodies followed by flow cytometry. Vero cells express both CD155 and CAR on the surface but do not express ICAM1. This is indicated by the percentage of PE-positive cells in the right part of each graph.

• 

  Figure 2. Expression of ICAM1 renders Vero cells susceptible to infection with HRV-B14. (a) Transient transfection of a plasmid encoding ICAM1 induces expression of ICAM1 in Vero cells as determined with immunohistochemistry; (b) infection of ICAM1 transiently transfected Vero cells with HRV-B14 induces replication of the virus as determined by the observation of CPE, whereas HRV-B14 cannot replicate in wild-type Vero cells. The upper row shows uninfected cells which function as negative control.

• 

  Figure 3. Level of ICAM1 on the surface of two stably transfected Vero clones. Expression of ICAM1 on the cell surface of two selected clones (Vero_ICAM1#4C4 and Vero_ICAM1#9G4), as determined by flow cytometric analyses. In red, staining with a PE-labelled isotype antibody and, in blue, staining with ICAM1-specific antibody. Both clones show ICAM1 expression, with highest expression in Vero_ICAM1#9G4. Percentages indicate proportion of cells expressing ICAM1.

• 

  Figure 4. Vero_ICAM1#4C4 cells stably expressing ICAM1 can be infected with HRV-B14, HRV-A16, HRV-A57 and HRV-B37. (a) Infection of Vero_ICAM1#4C4 and Vero_ICAM1#9G4 with HRV-B14 at an MOI of 0.1 induces replication in the ICAM1-expressing clones, but not in the parental Vero cells, indicated by the presence of CPE in both clones; (b) HRV-A16, HRV-A57 and HRV-B37 can replicate in Vero_ICAM1#4C4 cells. HRV-B14 and EV71_C4 were included as positive controls; (c) growth kinetics of HRV-B14 infection on Vero_ICAM1#4C4, Vero_ICAM1#9G4 and RD cells after infection at an MOI of 0.01.