Dong Xiao-Nan, Qi Yun, Ying Jian, Chen Xi, Chen Ying-Hua
Laboratory of Immunology, Department of Biology, Tsinghua University, Protein Science Laboratory of the Ministry of Education, Beijing 100084, PR China.
Vaccine. 2006 Jan 23;24(4):426-34. doi: 10.1016/j.vaccine.2005.08.002. Epub 2005 Aug 15.
Previously, two candidate multi-peptide-vaccines (MPVs) consisted of five overlapping synthetic peptides covering the antigenic domain B/C (aa693-777) on envelope protein E2 were prepared in our lab. And they successfully induced peptide-specific neutralizing antibodies and provided pigs with complete protection from the lethal challenge of virulent classical swine fever virus (CSFV) strain Shimen. In this study, these five peptides were conjugated to bovine serum albumin (BSA), with which five groups of pigs (n=10) were inoculated, respectively. Among these candidate peptide-vaccines (PVs), PV-BC1 (BC1: aa693-716) exhibited the most potent protective activity, PV-BC3, PV-BC4 and PV-BC5 (BC3: aa723-745; BC4: aa741-760; BC5: aa757-777) had weaker effects, while no effect of PV-BC2 (BC2: aa712-727) had been detected. Moreover, the polyclonal antibodies induced by PV-BC1 and PV-BC4 were capable of neutralizing C-strain virus in vitro. Thus, a principal sequential neutralizing determinant (aa693-716) and a minor sequential neutralizing determinant (aa741-760) were proved to lie in the antigenic domain B/C, which can be recruited into developing more effective "marker vaccine" by epitope-vaccine strategy. Our study also indicates that scanning with a panel of sequential peptide-immunogens is an effective method to locate sequential neutralizing epitopes.
此前,我们实验室制备了两种候选多表位疫苗(MPV),其由覆盖包膜蛋白E2上抗原结构域B/C(氨基酸693 - 777)的五条重叠合成肽组成。它们成功诱导了肽特异性中和抗体,并为猪提供了针对强毒经典猪瘟病毒(CSFV)石门株致死性攻击的完全保护。在本研究中,将这五条肽与牛血清白蛋白(BSA)偶联,分别接种五组猪(每组n = 10)。在这些候选肽疫苗(PV)中,PV - BC1(BC1:氨基酸693 - 716)表现出最强的保护活性,PV - BC3、PV - BC4和PV - BC5(BC3:氨基酸723 - 745;BC4:氨基酸741 - 760;BC5:氨基酸757 - 777)的效果较弱,而未检测到PV - BC2(BC2:氨基酸712 - 727)的效果。此外,由PV - BC1和PV - BC4诱导的多克隆抗体能够在体外中和C株病毒。因此,证明主要的序列中和决定簇(氨基酸693 - 716)和次要的序列中和决定簇(氨基酸741 - 760)位于抗原结构域B/C中,可通过表位疫苗策略用于开发更有效的“标记疫苗”。我们的研究还表明,用一组序列肽免疫原进行扫描是定位序列中和表位的有效方法。