Salehi-Sangani Ghodratollah, Mohebali Mehdi, Jajarmi Vahid, Khamesipour Ali, Bandehpour Mojgan, Mahmoudi Mahmoud, Zahedi-Zavaram Hadi
Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Department of Medical Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Iran J Basic Med Sci. 2019 Dec;22(12):1493-1501. doi: 10.22038/IJBMS.2019.14051.
To design a multivalent DNA vaccine encoding the most immunogenic regions of the antigens including TSA (Thiol-specific antioxidant protein), LmSTI1 ( stress-inducible protein1), LACK ( homologue of receptors for activated C Kinase), and KMP11 (kinetoplastid membrane protein-11) on BALB/c mice.
The chimeric construct was generated including the most immunogenic epitopes containing a combination of domains and oligopeptides of the aforementioned genes. The construct was cloned into pcDNA 3.1 plasmid and named "pleish-dom." Following intramuscular injection of mice, the capability of the vector pleish-dom alone and with pIL-12 (expressing murine IL-12) to raise protective cytokines and parasite burden was evaluated in the BALB/c mice as a susceptible animal model against .
The immunized mice with pleish-dom/pIL-12 showed the highest and the lowest levels of interferon-gamma (IFN-γ) and interleukin-10 (IL-10), as well as the lowest leishmanin skin test (LST) reactions, were found through 8 weeks post-infection.
Although the obtained DNA vaccine from the immunogenic chimeric protein of antigens was able to induce a high level of IFN-γ, it partially protected mice against L. major. However co-administration with pIL-12 led to shift immune response to Th1 phenotype, granuloma formation, and lowered parasite burden, and consequently distinct protection was found.
设计一种多价DNA疫苗,该疫苗编码抗原的最具免疫原性区域,这些抗原包括硫醇特异性抗氧化蛋白(TSA)、应激诱导蛋白1(LmSTI1)、活化C激酶受体同源物(LACK)以及动质体膜蛋白11(KMP11),并在BALB/c小鼠上进行实验。
构建嵌合结构,其包含上述基因的结构域和寡肽组合的最具免疫原性的表位。将该构建体克隆到pcDNA 3.1质粒中,并命名为“pleish-dom”。在对小鼠进行肌肉注射后,以BALB/c小鼠作为针对[病原体名称未给出]的易感动物模型,评估单独使用载体pleish-dom以及与表达小鼠IL-12的pIL-12联合使用时,引发保护性细胞因子和寄生虫负荷的能力。
在感染后8周内发现,用pleish-dom/pIL-12免疫的小鼠表现出最高和最低水平的干扰素-γ(IFN-γ)和白细胞介素-10(IL-10),以及最低的利什曼原虫皮肤试验(LST)反应。
尽管从抗原的免疫原性嵌合蛋白获得的DNA疫苗能够诱导高水平的IFN-γ,但它只能部分保护小鼠免受硕大利什曼原虫感染。然而,与pIL-12共同给药导致免疫反应向Th1表型转变、形成肉芽肿并降低寄生虫负荷,因此发现了明显的保护作用。