Dong Xiao-Nan, Wei Ke, Liu Zu-Qiang, Chen Ying-Hua
Department of Biology, Research Center for Medical Science, Tsinghua University, Beijing 100084, PR China.
Vaccine. 2002 Dec 13;21(3-4):167-73. doi: 10.1016/s0264-410x(02)00466-8.
Former investigations demonstrated that the envelope glycoprotein E2 could protect pigs from classical swine fever virus (CSFV). Based on these findings, we prepared synthetic peptide vaccine using E2 N-terminal antigenic units B/C and hoped to induce protective activity against lethal challenge of virulent CSFV strain Shimen. Five overlapped peptides sequence-covering amino acids 693-777 on E2 of Shimen were synthesized and then conjugated with bovine serum albumin (BSA), respectively. In the vaccination course, the candidate peptide vaccines in combination (multi-peptide vaccine (MPV)) were applied for immunization of pigs (n=10) and induced strong antibody response against CSFV. It is subsequently demonstrated that this peptide vaccine could provide immunized pigs complete protection against lethal CSFV challenge as C-strain does, while all non-immunized pigs in negative control group manifested obvious typical symptoms and died during the second and third weeks after viral challenge. In order to confirm the neutralizing activity of the polyclonal antibodies induced by MPV, neutralization assay were carried out on rabbits. The live C-strain alone could ordinarily induce typical fever on rabbits. The typical fever of rabbits induced by the live C-strain could be inhibited by pre-incubation with the anti-sera (dilution 1:4 and 1:16) induced by MPV, but not inhibited by pre-incubation with the same anti-sera from which the antibodies against five peptides were removed by peptide-specific affinity chromatography, which indicates that these peptide-specific antibodies in the anti-sera induced by MPV provided protective activity against CSFV. Our finding provides a new way to develop marker vaccine against CSFV.
先前的研究表明,包膜糖蛋白E2可保护猪免受经典猪瘟病毒(CSFV)感染。基于这些发现,我们使用E2 N端抗原单位B/C制备了合成肽疫苗,希望诱导针对强毒CSFV石门株致死性攻击的保护活性。合成了覆盖石门株E2上693 - 777位氨基酸的5个重叠肽序列,然后分别与牛血清白蛋白(BSA)偶联。在疫苗接种过程中,将候选肽疫苗联合应用(多肽疫苗(MPV))对猪(n = 10)进行免疫,诱导了针对CSFV的强烈抗体反应。随后证明,这种肽疫苗可像C株疫苗一样为免疫猪提供针对致死性CSFV攻击的完全保护,而阴性对照组中所有未免疫的猪在病毒攻击后的第二和第三周均表现出明显的典型症状并死亡。为了确认MPV诱导的多克隆抗体的中和活性,对兔子进行了中和试验。单独的活C株通常可使兔子产生典型发热。活C株诱导的兔子典型发热可被与MPV诱导的抗血清(稀释度1:4和1:16)预孵育所抑制,但不能被与通过肽特异性亲和层析去除针对5种肽的抗体后的相同抗血清预孵育所抑制,这表明MPV诱导的抗血清中的这些肽特异性抗体提供了针对CSFV的保护活性。我们的发现为开发针对CSFV的标记疫苗提供了新途径。