Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran.
Microb Pathog. 2023 Nov;184:106344. doi: 10.1016/j.micpath.2023.106344. Epub 2023 Sep 12.
Increasing evidence demonstrated that Enterohemorrhagic Escherichia coli (EHEC) and Shigella dysenteriae type 1 (S. dysenteriae1) are considered pathogens, that are connected with diarrhea and are still the greatest cause of death in children under the age of five years, worldwide. EHEC and S. dysenteriae 1 infections can be prevented and managed using a vaccination strategy against pathogen attachment stages. In this study, the chitosan nanostructures were loaded with recombinant EIT and STX1B-IpaD polypeptides. The immunogenic properties of this nano-vaccine candidate were investigated. The EIT and STX1B-IpaD recombinant proteins were heterologous expressed, purified, and confirmed by western blotting. The chitosan nanoparticles, were used to encapsulate the purified proteins. The immunogenicity of recombinant nano vaccine candidate, was examined in three groups of BalB/c mice by injection, oral delivery, and combination of oral-injection. ELISA and antibody titer, evaluated the humoral immune response. Finally, all three mice groups were challenged by two pathogens to test the ability of the nano-vaccine candidate to protect against bacterial infection. The Sereny test in guinea pigs was used to confirm the neutralizing effect of immune sera in controlling S. dysenteriae 1, infections. SDS-PAGE and western blotting, confirmed the presence and specificity of 63 and 27 kDa recombinant EIT and STX1B-IpaD, respectively. The results show that the nanoparticles containing recombinant proteins could stimulate the systemic and mucosal immune systems by producing IgG and IgA, respectively. The challenge test showed that, the candidate nano-vaccine could protect the animal model from bacterial infection. The combination of multiple recombinant proteins, carrying several epitopes and natural nanoparticles could evocate remarkable humoral and mucosal responses and improve the protection properties of synthetic antigens. Furthermore, compared with other available antigen delivery methods, using oral delivery as immune priming and injection as a booster method, could act as combinatorial methods to achieve a higher level of immunity. This approach could present an appropriate vaccine candidate against both EHEC and S. dysenteriae 1.
越来越多的证据表明，肠出血性大肠杆菌（EHEC）和志贺氏痢疾杆菌 1 型（S. dysenteriae1）被认为是病原体，与腹泻有关，仍是全球 5 岁以下儿童死亡的最大原因。可以使用针对病原体附着阶段的疫苗接种策略来预防和管理 EHEC 和 S. dysenteriae1 感染。在这项研究中，壳聚糖纳米结构负载了重组 EIT 和 STX1B-IpaD 多肽。研究了这种纳米疫苗候选物的免疫原性。异源表达、纯化了 EIT 和 STX1B-IpaD 重组蛋白，并通过 Western blot 进行了验证。壳聚糖纳米颗粒用于包封纯化的蛋白质。通过注射、口服和口服注射联合三种方式在三组 BalB/c 小鼠中检测重组纳米疫苗候选物的免疫原性。ELISA 和抗体滴度评估了体液免疫反应。最后，所有三组小鼠均受到两种病原体的攻击，以测试纳米疫苗候选物抵抗细菌感染的能力。豚鼠 Sereny 试验用于确认免疫血清在控制 S. dysenteriae1 感染中的中和作用。SDS-PAGE 和 Western blot 分别证实了 63 和 27 kDa 的重组 EIT 和 STX1B-IpaD 的存在和特异性。结果表明，含有重组蛋白的纳米颗粒通过产生 IgG 和 IgA 分别刺激系统和粘膜免疫系统。挑战试验表明，候选纳米疫苗可以保护动物模型免受细菌感染。多种携带多个表位的重组蛋白与天然纳米颗粒的结合可以引起显著的体液和粘膜反应，并提高合成抗原的保护特性。此外，与其他可用的抗原传递方法相比，口服传递作为免疫启动，注射作为增强方法，可以作为组合方法来实现更高水平的免疫。这种方法可以针对 EHEC 和 S. dysenteriae1 提供一种合适的疫苗候选物。
J Microbiol Methods. 2019-1-8
Iran J Allergy Asthma Immunol. 2021-8-7
Iran Biomed J. 2016
Artif Cells Nanomed Biotechnol. 2019-12
J Biomed Sci. 2025-7-8
Curr Issues Mol Biol. 2024-7-12
Zotero 插件