Mostaan Saied, Ghasemzadeh Abbas, Asadi Karam Mohammad Reza, Ehsani Parastoo, Sardari Soroush, Shokrgozar Mohammad Ali, Abolhassani Mohsen, Nikbakht Brujeni Gholamreza
Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran.
Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Vector Borne Zoonotic Dis. 2021 Nov;21(11):870-874. doi: 10.1089/vbz.2020.2758.
is the causative agent of a range of animal, and occasionally human, diseases. Problems with antimicrobial treatment of highlight the need to find other possible ways, such as prophylaxis, to manage infections. Current vaccines against include inactivated bacteria, live attenuated and nonpathogenic bacteria; they have disadvantages such as lack of immunogenicity, reactogenicity, or reversion to virulence. Using bioinformatics approaches, potentially immunogenic and protective epitopes were identified and merged to design the most optimally immunogenic triple epitope PlpE fusion protein of as a vaccine candidate. This triple epitope () was cloned into the pBAD/gIII A plasmid (pBR322-derived expression vectors designed for regulated, secreted recombinant protein expression and purification in ), expressed in Top 10 and purified in denatured form using Ni-NTA chromatography and 8 M urea. The immunogenicity of the purified proteins in BALB/c mice was assayed by measuring immunoglobulin G (IgG) responses. The protection potential was evaluated by challenging with 10 LD50 of serotype A:1, X-73 strain of and compared with commercially available inactivated fowl cholera vaccine and PlpE protein. IgG levels elicited by the polytope fusion protein of PlpE were higher than both commercially available inactivated fowl cholera vaccine and PlpE protein. Surprisingly, protection was independent of IgG level; commercially available inactivated fowl cholera vaccine (100% protection) was more protective than the polytope fusion protein (69% protection) and PlpE protein (69% protection). These results also confirm that IgG level is not a reliable indicator of protection. Further studies to evaluate the other antibody classes, such as immunoglobulin A or M, are required. The role of cell-mediated immunity should also be considered as a potential protection pathway.
是一系列动物疾病以及偶尔的人类疾病的病原体。针对[病原体名称]的抗菌治疗存在问题，这凸显了寻找其他可能方法（如预防）来控制感染的必要性。目前针对[病原体名称]的疫苗包括灭活细菌、减毒活细菌和非致病性细菌；它们存在诸如免疫原性不足、反应原性或毒力回复等缺点。利用生物信息学方法，鉴定并合并了潜在的免疫原性和保护性表位，以设计出作为疫苗候选物的最具免疫原性的[病原体名称]三重表位PlpE融合蛋白。将该三重表位（[具体名称]）克隆到pBAD/gIII A质粒（为在[具体宿主]中进行调控、分泌重组蛋白表达和纯化而设计的pBR322衍生表达载体）中，在Top 10 [宿主名称]中表达，并使用Ni-NTA色谱和8 M尿素以变性形式纯化。通过测量免疫球蛋白G（IgG）反应来检测纯化蛋白在BALB/c小鼠中的免疫原性。通过用10倍50%致死剂量的A:1血清型、[病原体名称]的X-73菌株进行攻毒来评估保护潜力，并与市售灭活禽霍乱疫苗和PlpE蛋白进行比较。[病原体名称]PlpE多表位融合蛋白引发的IgG水平高于市售灭活禽霍乱疫苗和PlpE蛋白。令人惊讶的是，保护作用与IgG水平无关；市售灭活禽霍乱疫苗（100%保护）比多表位融合蛋白（69%保护）和PlpE蛋白（69%保护）更具保护作用。这些结果也证实IgG水平不是保护作用的可靠指标。需要进一步研究来评估其他抗体类别，如免疫球蛋白A或M。细胞介导免疫的作用也应被视为一种潜在的保护途径。
