Sequential Cisplatin Therapy and Vaccination with HPV16 E6E7L2 Fusion Protein in Saponin Adjuvant GPI-0100 for the Treatment of a Model HPV16+ Cancer
Figure 2
Effect upon HPV16-specific T cell and neutralizing antibody responses to vaccination with TA-CIN/GPI-0100 after a freeze/thaw cycle.
A. Schematic illustration of the experimental protocol. Briefly, 5∼8 weeks old female BALB/c mice (5 mice/group) vaccinated subcutaneously with either 6.25 µg/mouse of TA-CIN, or formulated with 50 µg of GPI-0100, or formulated with 50 µg of GPI-0100 in 50.7 mg/mL of mannitol and subjected to a single freeze/thaw cycle. The mice were boosted twice with the same regimen with 2-week interval. Two weeks after the last vaccination, sera and splenocytes were harvested. B. Summary of the flow cytometric analysis of TA-CIN-specific CD4+ T cell responses analyzed by IFN-γ intracellular staining (and representative data is shown in S3A Fig.). C. Summary of the flow cytometry analysis examining HPV16-specific CD4+ T cell responses induced by TA-CIN formulated with GPI-0100 in mannitol and frozen/thawed once. Splenocytes were harvested after vaccination and stimulated with either TA-CIN or HPV16 L2 transfected CT26 cells (and representative data is shown in S3B Fig.). D. Summary of the flow cytometry analysis of TA-CIN-specific CD8+ T cell responses analyzed by IFN-γ intracellular staining. The data were acquired with FACSCalibur and analyzed with CellQuest (and representative data is shown in S3C Fig.). E. Summary of HPV16 L2-specific neutralizing antibody titer analyzed with HPV16-SEAP pseudovirus based neutralization assay.