A Dual-Type L2 11-88 Peptide from HPV Types 16/18 Formulated in Montanide ISA 720 Induced Strong and Balanced Th1/Th2 Immune Responses, Associated with High Titers of Broad Spectrum Cross-Reactive Antibodies in Vaccinated Mice

Figure 6

Generation and characterization of HPV 16 and HPV 18 pseudovirions. Two hundred ninety-three TT cells (2 × 10⁷) were cultured in 162 cm² flasks. Twenty hours after culture, the cells were transfected with 40 μg of either p16L1L2 or p18L1L2 plasmids and 40 μg of the GFP-expressing reporter plasmid (pfwB). Generation of pseudovirions was confirmed by transducing 293 TT cells using crude extract cell lysates at a 1 : 100 dilution in 24-well plates in triplicate for each PsV. Forty-eight hours after infection, cells expressing GFP were analyzed by flow cytometry (a and b) and observed under inverted microscope (c). Furthermore, crude cell extract lysates containing PsV 16 were also subjected to transmission electron microscopy (visualized at a magnification of 50,000) to confirm particles (d).

(a)

(b)

(c)

(d)