Figure 1.
Assessment of infectivity, L2 incorporation, and reporter DNA encapsidation by PsV preparations of 34 HPV types.
(A) 250 million particles (as estimated based on L1 concentration) of HPV PsV were transferred to 293TT cells, incubated at 37°C for 72 hours before cell lysis. Luciferase activity was measured in cell lysate (n = 4). (B) Western blot analysis detecting L2 in 100 ng of each HPV PsV preparation using rabbit antiserum to L2 α11-88x8. (C) Reporter plasmid copy number of each PsV preparations for each HPV type sample was measured by quantitative real time PCR. Black and blue bars represent alpha and beta type, respectively, and mean ± standard error plotted.
Table 1.
Summary of the phylogeny, tropism, charge and in vitro neutralization by L2 11-88x8 antiserum of the 34 HPV genotypes tested.
Figure 2.
Inhibition of HPV PsV infection by titrations of heparin and carrageenan.
PsVs of each HPV type carrying a luciferase reporter gene were incubated with 1000 µg/mL, 100 µg/mL, 10 µg/ml or 0 µg/ml heparin (A) or carrageenan (B) at 37°C for 1 hour. The mixtures were transferred to 293TT cells for 72 h. After incubation cells were lysed and luciferase activity was measured, and percent infection compared to control was calculated (n = 3). The percent inhibition of infection by each HPV PsV in the presence of 100 µg/mL of heparin (C) or carrageenan (D) was also plotted separately. Red and blue bars and lines represent mucosal and cutaneous HPV types, respectively.
Figure 3.
Impact of furin inhibitor on infection by PsV of diverse HPV.
PsVs of each indicated HPV type carrying a luciferase reporter gene were transferred to 293TT cells for 72 hours in the presence or absence of 20 µM furin inhibitor. After incubation, cells were lysed, luciferase activity was measured and percent inhibition of infectivity compared to control calculated. Red and blue bars represent mucosal and cutaneous HPV types, respectively.
Figure 4.
Impact of γ-secretase inhibitor on infection by PsV of diverse HPV.
PsVs of each indicated HPV type carrying a luciferase reporter gene were transferred to 293TT cells for 72γ-secretase inhibitor XXI (n = 3). After incubation, luciferase activity was measured and percent inhibition of infectivity compared to control calculated. Red and blue bars represent mucosal and cutaneous HPV types, respectively.