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在小鼠模型中,使用含有重组触发因子/Bp26/Omp31嵌合蛋白的非离子表面活性剂囊泡和甘露糖基化非离子表面活性剂囊泡进行鼻内和腹腔内免疫以预防感染。

Intranasal and intraperitoneal immunization against infection using niosome and mannosylated niosomes containing recombinant trigger factor/Bp26/Omp31 chimeric protein in a mouse model.

作者信息

Sharif Fahimeh, Nazari Razieh, Fasihi-Ramandi Mahdi, Taheri Ramezan Ali, Zargar Mohsen

机构信息

Department of Microbiology, Islamic Azad University Qom Branch, Qom, Iran.

Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

出版信息

Clin Exp Vaccine Res. 2024 Jul;13(3):232-241. doi: 10.7774/cevr.2024.13.3.232. Epub 2024 Jul 31.

Abstract

PURPOSE

Brucellosis, a zoonotic infectious disease, is a worldwide health issue affecting animals and humans. No effective human vaccine and the complications caused by the use of animal vaccines are among the factors that have prevented the eradication of the disease worldwide. However, bio-engineering technologies have paved the way for designing new targeted and highly efficacious vaccines. In this regard, the study aimed to evaluate immunity induced by mannosylated niosome containing recombinant trigger factor/Bp26/Omp31 (rTBO) chimeric protein in a mouse model.

MATERIALS AND METHODS

rTBO as chimeric antigen (Ag) was expressed in BL21 (DE3) and, after purification, loaded on niosome and mannosylated niosome. The characteristics of the nanoparticles were assessed. The mice were immunized using rTBO, niosome, and mannosylated niosome-rTBO in intranasal and intraperitoneal routes. Serum antibodies (immunoglobulin [Ig]A, IgG, IgG1, and IgG2a) and splenocyte cytokines (interferon-gamma, interleukin [IL]-4, and IL-12) were evaluated in immunized mice. Finally, immunized mice were challenged by and . A high antibody level was produced by niosomal antigen (Nio-Ag) and mannosylated noisomal antigen (Nio-Man-Ag) compared to the control after 10, 24, and 38 days of immunization. The IgG2a/IgG1 titer ratio for Nio-Man-Ag was 1.2 and 1.1 in intraperitoneal and intranasal methods and lower than one in free Ag and Nio-Ag. Cytokine production was significantly higher in the immunized animal with Ag-loaded nanoparticles than in the negative control group (p<0.05). Moreover, cytokine and antibody levels were significantly higher in the injection than in the inhalation method (p<0.05).

RESULTS

The combination of mannosylated noisome and rTBO chimeric proteins stimulate the cellular and humoral immune response and produce cytokines, playing a role in developing the protective acquired immune response in the infectious model. Also, the intraperitoneal route resulted in a successful enhancement of cytokines production more than intranasal administration.

CONCLUSION

Designing an effective vaccine candidate against that selectively induces cellular and humoral immune response can be done by selecting a suitable nanoniosome formulation as an immunoadjuvant and recombinant protein as an immune response-stimulating Ag.

摘要

目的

布鲁氏菌病是一种人畜共患传染病,是影响动物和人类的全球性健康问题。缺乏有效的人类疫苗以及动物疫苗使用引起的并发症是阻碍全球根除该疾病的因素之一。然而,生物工程技术为设计新型靶向高效疫苗铺平了道路。在这方面,本研究旨在评估含重组触发因子/Bp26/Omp31(rTBO)嵌合蛋白的甘露糖基化脂质体在小鼠模型中诱导的免疫反应。

材料与方法

rTBO作为嵌合抗原(Ag)在BL21(DE3)中表达,纯化后负载于脂质体和甘露糖基化脂质体上。评估纳米颗粒的特性。小鼠通过鼻内和腹腔内途径用rTBO、脂质体和甘露糖基化脂质体-rTBO进行免疫。检测免疫小鼠的血清抗体(免疫球蛋白[Ig]A、IgG、IgG1和IgG2a)和脾细胞细胞因子(干扰素-γ、白细胞介素[IL]-4和IL-12)。最后,用……对免疫小鼠进行攻毒。与对照组相比,免疫10、24和38天后,脂质体抗原(Nio-Ag)和甘露糖基化脂质体抗原(Nio-Man-Ag)产生了高水平抗体。Nio-Man-Ag的IgG2a/IgG1滴度比在腹腔内和鼻内方法中分别为1.2和1.1,低于游离Ag和Nio-Ag中的比值。负载Ag的纳米颗粒免疫的动物中细胞因子产生显著高于阴性对照组(p<0.05)。此外,注射组的细胞因子和抗体水平显著高于吸入组(p<0.05)。

结果

甘露糖基化脂质体与rTBO嵌合蛋白的组合刺激细胞和体液免疫反应并产生细胞因子,在感染模型中对保护性获得性免疫反应的发展起作用。而且,腹腔内途径比鼻内给药更成功地增强了细胞因子的产生。

结论

通过选择合适的纳米脂质体制剂作为免疫佐剂和重组蛋白作为刺激免疫反应的Ag,可以设计出一种针对……的有效候选疫苗,选择性地诱导细胞和体液免疫反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3942/11319111/128ec9a61691/cevr-13-232-g001.jpg

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