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hep26668-sup-0001-suppfigure1.tif3047KsFigure 1. Establishment of an HBV persistent mouse model. (A) C57BL/6 mice were hydrodynamically injected with 6 μg of either pAAV/HBV1.2 or pAAV/control plasmid. Serum HBsAg titers were measured at the indicated time points using immunoradiometric assay (IRMA). (B) The positive detection rate of serum HBsAg levels at different time points in mice that received 6 μg of HBV plasmid. (C) Correlation between serum HBsAg titer and HBV DNA copy number. HBV DNA copy number was determined using quantitative PCR. (D) Liver sections were processed for HBcAg expression and (E) H&E staining at 6 wpi (original magnification: 100×).
hep26668-sup-0002-suppfigure2.tif1934KsFigure 2. Pre-existing anti-HBV immunity prevents HBV persistence in the liver. (A) C57BL6 mice were immunized intramuscularly with either 1 μg of HBV vaccine or control PBS within a 2-week interval; 1 week later, they were hydrodynamically injected with HBV plasmid as described for establishing HBV-carrier mice. Serum samples were collected and analyzed for HBsAg (B) and HBeAg (C) by IRMA at the indicated time points. (D) Immunohistochemical staining for HBcAg in the livers of HBsAg- or PBS-immunized mice at 6 wpi (original magnification: 100×). Data are expressed as mean ± SEM (n = 10 mice/group) and represent 3 independent experiments that gave similar results.
hep26668-sup-0003-suppfigure3.tif766KsFigure 3. Reconstitution of adaptive immune cells after cellular transfer. Splenocytes were isolated from HBV-carrier and control mice and were transferred intravenously (5 × 107) into Rag1-/- recipient mice. T and B cell percentages were shown 1 week after splenocyte transfer as determined by FACS analysis.
hep26668-sup-0004-suppfigure4.tif796KsFigure 4. Spleoncytes from HBsAg vaccinated mice decrease serum HBsAg and HBeAg levels in HBV+Rag1-/- recipient mice. Donor mice were immunized with HBsAg vaccine or PBS as control. Donor cells were divided into 4 parts: total splenocytes from control mice; total splenocytes from HBsAg vaccinated mice (anti-HBV); CD19+ cells of splenocytes purified by MACS from HBsAg vaccinated mice; and CD19- cells of splenocytes from HBsAg vaccinated mice. These cells were transferred into HBV+Rag1-/- mice respectively. Two weeks after cells transfer, the serum levels of HBsAg (A) and HBeAg (B) were detected in recipient mice. Data are expressed as mean ± SEM. The numbers above bars in Fig. B mean the number of HBeAg negative mice within total recipient mice.

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