Figure 6.

The hepatitis B virus e antigen (HBeAg) -specific double-negative (DN) T-cell population originates from DN progenitor T cells. HBeAg × 7/16-5 double transgenic (dbl-Tg) splenocytes were harvested and prepared as single cell suspensions. The CD4⁺, CD8⁺ or both CD4⁺/CD8⁺ T cells were depleted, respectively, using magnetic beads, and flow through from each preparation was cultured. For the progenitor DN-depleted culture, positively purified CD4⁺ and CD8⁺ cells were cultured with antigen-presenting cells. The HBeAg-specific DN progenitor cells were depleted as described in the Materials and methods. Each preparation of cells was cultured in the presence of 2 μg/ml of p120–140 for 4 days. Cells were gated for DN (CD4⁻ and CD8⁻) and further analysed for Vβ11 positivity.