Fig. 5

CpG-ODN-induced monocyte mobilization regulates TVGV-HBx-mediated HBV clearance. a-b HBV carrier mice (N = 3) received TVGV-HBx on day 0. Gemcitabine (40 mg/kg) was given by intraperitoneal injection on days 0 and 1. CLs (200 μL) were given by intravenous injection on day 0. PBMCs were isolated on day 2. PBMCs were stained with fluorochrome-conjugated anti-CD11b, anti-CD115, anti-Ly6C and anti-Ly6G antibodies and then subjected to FACS analysis. a Frequency of CD11b⁺CD115⁺Ly6G⁻ monocytes among total PBMCs. b Representative FACS plots of Ly6C⁺ monocyte subsets (gated by CD11b + CD115 + Ly6G-) before and after vaccination and drug administration. The cell frequencies of each subpopulation are shown on the plot. c-d HBV carrier mice (N = 4 ~ 5) were administered TVGV-HBx vaccination (black arrow with the dosage indicated, 100 μg of antigen plus 20 μg of CpG-ODN as 1.0x) and a monocyte-depleting drug (red arrow) at the indicated time points. Blood samples were collected at the indicated time points. c Serum HBs titer after TVGV-HBx vaccination and CL treatment. d Serum HBs titer after TVGV-HBx vaccination and gemcitabine treatment. e Splenocyte IFN-γ ELISpot results (mice N = 5) after TVGV-HBx vaccination and gemcitabine treatment. The vaccination and drug administration protocol were the same as in (d), and the splenocytes were harvested on day 14. Cells were stimulated with HBx-derived 15-mer overlapping peptides. Statistics: Student’s t-test. *, p < 0.05; **, p < 0.01; NS, not significant; LoD, limit of detection; SFCs, spot-forming cells; GEM, gemcitabine