Fig. 4.

GC B cells and Tfh cells are low in the DLN of HBV-carrier mice after HBsAg vaccination. At 12 d after HBsAg vaccination at the tibialis anterior muscle, lymphocytes from the DLN (popliteal LN) were stained with different antibodies. (A) The DLN was photographed (Left) and total cells were counted (Right). (B) DLN sections were stained with Biotin-PNA; GCs are shown (Scale bar, 200 μm). (C) GC B cell (B220⁺Gl-7⁺Fas⁺) frequency and (D) Bcl-6 expression levels were examined. The MNC amount of the popliteal LN was too low from control mice without immunization with HBsAg (red symbol), so we put MNCs from two mice into one tube for intracellular staining by flow cytometry. (E) Tfh cell (CD4⁺CXCR5⁺PD-1⁺) frequency and (F) CD44⁺ ICOS⁺ cell percentage among CD4⁺ T cells were measured. Results represent three independent experiments (n = at least 3 per group). *P < 0.05, **P < 0.01, and ***P < 0.001.