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展示口蹄疫病毒(FMDV)G-H环的T7噬菌体纳米颗粒的免疫原性

Immunogenicity of T7 bacteriophage nanoparticles displaying G-H loop of foot-and-mouth disease virus (FMDV).

作者信息

Xu Hai, Bao Xi, Lu Yu, Liu Yamei, Deng Bihua, Wang Yiwei, Xu Yue, Hou Jibo

机构信息

National Research Center of Engineering and Technology for Veterinary Biologicals, Jiangsu Academy of Agricultural Science, Nanjing 210014, Jiangsu Province, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, Jiangsu Province, China.

National Research Center of Engineering and Technology for Veterinary Biologicals, Jiangsu Academy of Agricultural Science, Nanjing 210014, Jiangsu Province, China.

出版信息

Vet Microbiol. 2017 Jun;205:46-52. doi: 10.1016/j.vetmic.2017.04.023. Epub 2017 May 4.

Abstract

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals that causes severe economic losses worldwide. The G-H loop of the FMDV VP1 structural protein is the major neutralizing antigenic site. However, a fully protective G-H loop peptide vaccine requires the addition of promiscuous Th sites from a source outside VP1. Thus, we demonstrated the potential of T7 bacteriophage based nanoparticles displaying a genetically fused G-H loop peptide (T7-GH) as a FMDV vaccine candidate. Recombinant T7-GH phage was constructed by inserting the G-H loop coding region into the T7 Select 415-1b vector. Purified T7-GH phage nanoparticles were analyzed by SDS-PAGE, Western blot and Dot-ELISA. Pigs seronegative for FMDV exposure were immunized with T7-GH nanoparticles along with the adjuvant Montanide ISA206, and two commercially available FMDV vaccines (InactVac and PepVac). Humoral and cellular immune responses, as well as protection against virulent homologous virus challenge were assessed following single dose immunization. Pigs immunized T7-GH developed comparable anti-VP1 antibody titers to PepVac, although lower LPBE titers than was induced by InactVac. Antigen specific lymphocyte proliferation was detected in T7-GH group similar to that of PepVac group, however, weaker than InactVac group. Pigs immunized with T7-GH developed a neutralizing antibody response stronger than PepVac, but weaker than InactVac. Furthermore, 80% (4/5) of T7-GH immunized pigs were protected from challenge with virulent homologous virus. These findings demonstrate that the T7-GH phage nanoparticles were effective in eliciting antigen specific immune responses in pigs, highlighting the value of such an approach in the research and development of FMDV vaccines.

摘要

口蹄疫(FMD)是偶蹄动物的一种高度传染性疾病,在全球范围内造成严重经济损失。口蹄疫病毒(FMDV)VP1结构蛋白的G-H环是主要的中和抗原位点。然而,一种完全保护性的G-H环肽疫苗需要添加来自VP1以外来源的通用Th表位。因此,我们证明了展示基因融合G-H环肽(T7-GH)的基于T7噬菌体的纳米颗粒作为FMDV疫苗候选物的潜力。通过将G-H环编码区插入T7 Select 415-1b载体构建重组T7-GH噬菌体。通过SDS-PAGE、蛋白质免疫印迹和斑点酶联免疫吸附测定法分析纯化的T7-GH噬菌体纳米颗粒。将未接触过FMDV的血清阴性猪用T7-GH纳米颗粒与佐剂Montanide ISA206以及两种市售FMDV疫苗(灭活疫苗和肽疫苗)一起免疫。在单剂量免疫后评估体液和细胞免疫反应以及针对强毒同源病毒攻击的保护作用。用T7-GH免疫的猪产生的抗VP1抗体效价与肽疫苗相当,尽管低聚肽结合酶(LPBE)效价比灭活疫苗诱导的低。在T7-GH组中检测到的抗原特异性淋巴细胞增殖与肽疫苗组相似,但弱于灭活疫苗组。用T7-GH免疫的猪产生的中和抗体反应强于肽疫苗,但弱于灭活疫苗。此外,80%(4/5)用T7-GH免疫的猪在受到强毒同源病毒攻击时得到保护。这些发现表明,T7-GH噬菌体纳米颗粒在猪中能有效引发抗原特异性免疫反应,突出了这种方法在FMDV疫苗研发中的价值。

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