Construction and characterization of an experimental ISCOMS-based hepatitis B polypeptide vaccine

AIM: To characterize the biochemical and immunological properties of an experimental ISCOMS vaccine prepared from a novel therapeutic polypeptide based on T cell epitopes of HBsAg, and a heptatis B-ISCOMS was prepared and investigated. METHODS: An immunostimulating complexes(ISCOMS)-based vaccine containing a novel therapeutic hepatitis B polypeptide was prepared by dialysis method, and its formation was visualized by electron microscopy and biochemically verified by SDS-polyacrylamide gel electrophoresis. Amount of the peptide within ISCOMS was determined by Bradford assay, and specific CTL response was detected by ELISPOT assay. RESULTS: Typical cage-like structures of submicroparticle with a diameter of about 40nm were observed by electron microscopy. Results from Bradford assay showed that the level of peptide incorporation was about 0.33g.L(-1). At the paralleled position close to the sixth band of the molecular weight marker(3480kDa) a clear band was shown in SDS-PAGE analysis, indicating successful incorporation of polypeptide into ISCOMS. It is suggested that ISCOMS delivery system could efficiently improve the immunogenicity of polypeptide and elicit specific immune responses in vivo by the results of ELISPOT assay, which showed that IFN-gamma producing cells(specific CTL responses) were increased(spots of ISCOMS-treated group: 47+/-5, n =3; control group: 5+/-2, n =3). CONCLUSION: ISCOMS-based hepatitis B polypeptide vaccine is successfully constructed and it induces a higher CTL response compared with short polypeptides vaccine in vivo.

瞄准：描绘生物化学并且基于 HBsAg 的 T 房间 epitopes 从新奇治疗学的多肽准备的一支试验性的 ISCOMS 疫苗的免疫学的性质，和 heptatis B-ISCOMS 被准备并且调查。方法：刺激建筑群(ISCOMS ) 的免疫基于疫苗的包含新奇治疗学的肝炎 B 多肽被分离方法准备，并且它的形成被 SDS-polyacrylamide 胶化电气泳动化学上验证的电子显微镜学和简历设想。在 ISCOMS 以内的肽的数量被 Bradford 试金决定，并且特定的 CTL 反应被 ELISPOT 试金检测。结果：有大约 40nm 的一条直径的 submicroparticle 的典型像笼子的结构被电子显微镜学观察。从 Bradford 试金的结果证明肽加入的水平关于 0.33g .L (-1) 。在到分子的重量标记(3480kDa ) 的第六个乐队的 paralleled 位置结束，一个清楚的乐队在 SDS 页分析被给看，显示进 ISCOMS 的多肽的成功的加入。ISCOMS 交货系统能高效地改进多肽的 immunogenicity 并且由 ELISPOT 试金的结果在 vivo 得到特定的有免疫力的回答，这被建议，它证明生产房间(特定的 CTL 回答) 的 IFN-gamma 被增加(对待 ISCOMS 的组的点：47+/-5， n =3；控制组：5+/-2， n =3 ) 。结论：基于 ISCOMS 的肝炎 B 多肽疫苗成功地与在 vivo 疫苗的短多肽相比被构造，它导致更高的 CTL 回答。