利用重叠的preS1片段融合蛋白鉴定乙肝表面抗原（HBsAg）preS1区域中的免疫原性结构域。
Identification of the immunogenic domains in HBsAg preS1 region using overlapping preS1 fragment fusion proteins.
作者信息
Hu Wei-Guo, Wei Jun, Xia Heng-Chuan, Yang Xin-Xiu, Li Feng, Li Guang-Di, Wang Yuan, Zhang Zu-Chuan
机构信息
Key Laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue-yang Road, Shanghai 200031, China.
出版信息
World J Gastroenterol. 2005 Apr 14;11(14):2088-94. doi: 10.3748/wjg.v11.i14.2088.
AIM
The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purpose was to further investigate its immunogenic domains for the epitope-based hepatitis B vaccine design.
METHODS
Eight GST fusion proteins containing overlapping preS1 fragments in preS1 (21-119) region were expressed in E.coli. Using these purified fusion proteins, the immunogenic domains in preS1 region were identified in detail in mice and humans by Western blot analysis and ELISA.
RESULTS
The results in mice showed that the immu-nogenic domains mainly existed in preS1 (21-59) and preS1 (95-109). Similarly, these fragments had strong immunogenicity in humans; whereas the other parts except for preS1 (60-70) also had some immunogenicity. More importantly, a major immunogenic domain, preS1 (34-59), which has much stronger immunogenicity, was identified. Additionally, the antibodies against some preS1 fragments, especially preS1 (34-59), were speculated to be virus-neutralizing.
CONCLUSION
Eight GST fusion proteins containing overlapping preS1 fragments were prepared successfully. They were used for the study on the immunogenic dom-ains in preS1 (21-119) region. The preS1 (34-59) fragments were the major immunogenic domains in the preS1 region, and the antibodies against these fragments were speculated to be virus-neutralizing. Therefore, the incorporation of preS1 (34-59) fragments into epitope-based HBV vaccines may be efficient for enhancement of immune response. Additionally, the results also imply that there are more complex immune responses to preS1 region and more abundant immunogenic domains in humans.
目的
将乙肝病毒（HBV）前S1区纳入基于表位的乙肝疫苗已被广泛接受，但前S1序列的掺入位点和大小存在争议。因此，我们的目的是进一步研究其免疫原性结构域，以用于基于表位的乙肝疫苗设计。
方法
在大肠杆菌中表达了8种在preS1（21-119）区域包含重叠preS1片段的GST融合蛋白。使用这些纯化的融合蛋白，通过蛋白质印迹分析和酶联免疫吸附测定在小鼠和人类中详细鉴定了preS1区域的免疫原性结构域。
结果
小鼠实验结果表明，免疫原性结构域主要存在于preS1（21-59）和preS1（95-109）。同样，这些片段在人类中具有较强的免疫原性；而除preS1（60-70）外的其他部分也具有一定的免疫原性。更重要的是，鉴定出了一个主要的免疫原性结构域preS1（34-59），其免疫原性更强。此外，推测针对某些preS1片段，尤其是preS1（34-59）的抗体具有病毒中和作用。
结论
成功制备了8种包含重叠preS1片段的GST融合蛋白。它们用于研究preS1（21-119）区域的免疫原性结构域。preS1（34-59）片段是preS1区域的主要免疫原性结构域，推测针对这些片段的抗体具有病毒中和作用。因此，将preS1（34-59）片段纳入基于表位的乙肝疫苗可能有效增强免疫反应。此外，结果还表明，人类对preS1区域存在更复杂的免疫反应，免疫原性结构域更丰富。
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