Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran.
Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
J Biotechnol. 2020 Feb 20;310:89-96. doi: 10.1016/j.jbiotec.2020.01.016. Epub 2020 Feb 1.
Brucellosis is a worldwide bacterial zoonosis disease. Live attenuated Brucella vaccines have several drawbacks. Thus development of a safe and effective vaccine for brucellosis is a concern of many scientists. FliC protein contributes in virulence of Brucella; hence, it is a promising target for brucellosis vaccine. In this study, Mannosylated Chitosan Nanoparticles (MCN) loaded with FliC protein were synthesized as a targeted vaccine delivery system. The immunogenicity and protective efficacy of FliC and FliC-MCN against Brucella infection were evaluated in BALB/c mice. After cloning, expression and purification, FliC protein was loaded on MCN. The particle size, loading efficiency and in vitro release of the NPs were determined. Our investigation revealed that FliC and FliC-MCN could significantly increase specific IgG response (higher IgG2a titers). Besides, spleen cells from immunized mice produced high level of IFN-γ and IL-2 and low level IL-10 cytokines. Immunization with FliC and FliC-MCN conferred significant degree of protection against B. melitensis 16 M and B. abortus 544 infections. Overall these results indicate that FliC protein would be a novel potential antigen candidate for the development of a subunit vaccine against B. melitensis and B. abortus. Moreover, MCN could be used as an adjuvant and targeted vaccine delivery system.
布鲁氏菌病是一种全球性的细菌性人畜共患病。活减毒布鲁氏菌疫苗有几个缺点。因此，开发一种安全有效的布鲁氏菌疫苗是许多科学家关注的问题。FliC 蛋白有助于布鲁氏菌的毒力；因此，它是布鲁氏菌疫苗的一个有前途的靶点。在这项研究中，载有 FliC 蛋白的甘露聚糖化壳聚糖纳米颗粒（MCN）被合成作为一种靶向疫苗传递系统。在 BALB/c 小鼠中评估了 FliC 和 FliC-MCN 对布鲁氏菌感染的免疫原性和保护效力。在克隆、表达和纯化后，将 FliC 蛋白加载到 MCN 上。测定了 NPs 的粒径、载药量和体外释放。我们的研究表明，FliC 和 FliC-MCN 可以显著增加特异性 IgG 反应（更高的 IgG2a 滴度）。此外，免疫小鼠的脾细胞产生高水平的 IFN-γ和 IL-2，以及低水平的 IL-10 细胞因子。用 FliC 和 FliC-MCN 免疫可显著预防 B. melitensis 16M 和 B. abortus 544 感染。总之，这些结果表明 FliC 蛋白将是开发针对 B. melitensis 和 B. abortus 的亚单位疫苗的一种新型潜在抗原候选物。此外，MCN 可用作佐剂和靶向疫苗传递系统。
