■布鲁氏菌病,一种人畜共患传染病,是影响动物和人类的世界性健康问题。没有有效的人类疫苗和使用动物疫苗引起的并发症是阻止全世界根除该疾病的因素之一。然而,生物工程技术为设计新的靶向和高效疫苗铺平了道路。在这方面,本研究旨在评估含有布鲁氏菌重组触发因子/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力。
■rTBO作为嵌合抗原(Ag)在大肠杆菌BL21(DE3)中表达,净化后,装载在niosome和甘露糖基化niosome上。评估纳米颗粒的特性。使用rTBO免疫小鼠,niosome,鼻内和腹膜途径的甘露糖基化niosome-rTBO。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,在免疫小鼠中评估白介素[IL]-4和IL-12)。最后,用B.melitensis和B.abortus攻击免疫小鼠。免疫10、24和38天后,与对照相比,通过脂质体抗原(Nio-Ag)和甘露糖基化的脂质体抗原(Nio-Man-Ag)产生了较高的抗体水平。在腹膜内和鼻内方法中,Nio-Man-Ag的IgG2a/IgG1滴度比为1.2和1.1,而在游离Ag和Nio-Ag中低于1。在具有负载Ag的纳米颗粒的免疫动物中,细胞因子产生显著高于阴性对照组(p<0.05)。此外,注射方法中细胞因子和抗体水平显著高于吸入方法(p<0.05)。
■甘露糖基化噪声体和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,在布鲁氏菌感染模型中发挥保护性获得性免疫应答的作用。此外,与鼻内给药相比,腹膜内途径成功增强了细胞因子的产生。
设计选择性诱导细胞和体液免疫应答的针对布鲁氏菌的有效疫苗候选物可以通过选择合适的纳米体制剂作为免疫佐剂和重组蛋白作为免疫应答刺激Ag来完成。
UNASSIGNED: 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 Brucella recombinant trigger factor/Bp26/Omp31 (rTBO) chimeric protein in a mouse model.
UNASSIGNED: rTBO as chimeric antigen (Ag) was expressed in Escherichia coli 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 B. melitensis and B. abortus. 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).
UNASSIGNED: 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 Brucella infectious model. Also, the intraperitoneal route resulted in a successful enhancement of cytokines production more than intranasal administration.
UNASSIGNED: Designing an effective vaccine candidate against Brucella 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.