Viral Hepatitis
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 21, 2005; 11(19): 2858-2863
Published online May 21, 2005. doi: 10.3748/wjg.v11.i19.2858
Enhancement of humoral immune responses to HBsAg by heat shock protein gp96 and its N-terminal fragment in mice
Hong-Tao Li, Jia-Bin Yan, Jing Li, Ming-Hai Zhou, Xiao-Dong Zhu, Yu-Xia Zhang, Po Tien
Hong-Tao Li, Jia-Bin Yan, Ming-Hai Zhou, Xiao-Dong Zhu, Yu-Xia Zhang, Po Tien, Department of Molecular Virology, Institute of Microbiology, Graduate School of the Chinese Academy of Sciences, Beijing 100080, China
Jing Li, Hepatitis B Department, National Vaccine and Serum Institute, Beijing 100024, China
Author contributions: All authors contributed equally to the work.
Supported by the Major State Basic Research Development Program of China, Program 973, Grant No. 2001CB510001
Correspondence to: Professor Po Tien, Department of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Zhongguancun Beiyitiao, Beijing 100080, China. tienpo@sun.im.ac.cn
Telephone: +86-10-62554247 Fax: +86-10-62622101
Received: June 11, 2004
Revised: June 12, 2004
Accepted: June 29, 2004
Published online: May 21, 2005
Abstract

AIM: Most studies on the immune effect of gp96 were focused on its enhancement of CTLs. It is interesting to know whether gp96 could influence the humoral immune response, and whether the recombinant N-terminal fragment of gp96 could substitute native gp96 to stimulate the immune system.

METHODS: gp96 isolated from livers of normal mice and its N-terminal fragment (amino acid 22-355) expressed in E coli were used for immunization of BALb/c mice. Eight groups of mice received one of the following regiments subcutaneously in 100 μL phosphate buffered saline (PBS) at an interval of 3 wk. Group 1: PBS only; group 2: gp96 only; group 3: N-terminal fragment only; group 4: HBsAg only; group 5: HBsAg+gp96; group 6: HBsAg+N-terminal fragment; group 7: HBsAg+incomplete Freud’s adjuvant; group 8: HBsAg+N-terminal fragment (95 °C heated for 30 min). Serum anti-HBsAg antibody levels were assayed by ELISA. CTL responses in splenocytes were analyzed by ELISPOT after the last vaccination.

RESULTS: The average titer of serum anti-HBsAg antibody in the mice immunized with HBsAg together with gp96 or its N-terminal fragment were much higher than those immunized with HBsAg alone detected by ELISA. The cellular immune response of the mice immunized with HBsAg together with gp96 or its N-terminal fragment was not different with those immunized with HBsAg alone measured by ELISPOT assay.

CONCLUSION: gp96 or its N-terminal fragment greatly improved humoral immune response induced by HBsAg, but failed to enhance the CTL response, which demonstrated the potential of using gp96 or its N-terminal fragment as a possible adjuvant to augment humoral immune response against HBV infection.

Keywords: Heat shock protein; gp96 N-terminal fragment; HBV; Hepatitis B virus surface antigen (HBsAg); Vaccine