Enhancement of humoral immune responses to HBsAg by heat shock protein gp96 and its N-terminal fragment in mice

Figure 3 Induced humoral immune response in Balb/c mice. A: Groups (eight mice per group) of Balb/c mice were immunized with different gradient: group 1, PBS only; group 2, 10 μg gp96 only; group 3, 10 μg NTF only; group 4, 10 μg HBsAg only; group 5, 10 μg HBsAg+10 μg NTF; group 6, 10 μg HBsAg+10 μg gp96; group 7, 10 μg HBsAg+IFA; group 8, 10 μg HBsAg+10 μg NTF (95 °C heated for 30 min) at d 0 (day of first vaccination). At d 21, and 42 mice were re-immunized with the same material. At d 63 anti-HBsAg antibodies from different groups were compared using the value at 450 nm absorbance. The values of antibody were expressed as geometry mean±SD; B: Sera from the mice immunized with HBsAg plus heated NTF, HBsAg plus NTF or gp96 (groups 5, 6 and 8) were collected on d 14, 35 and 64 from first vaccination and stored at -20 °C and titers at different dilution were tested at the same time.