Wang Hai-Long, Zhang Tie-E, Yin Li-Tian, Pang Min, Guan Li, Liu Hong-Li, Zhang Jian-Hong, Meng Xiao-Li, Bai Ji-Zhong, Zheng Guo-Ping, Yin Guo-Rong
Research Institute of Medical Parasitology, Shanxi Medical University, Taiyuan, Shanxi, PR China.
Department of Physiology, Key Laboratory of Cellular Physiology Co-constructed by Province and Ministry of Education, Shanxi Medical University, Taiyuan, Shanxi, PR China.
PLoS One. 2014 Sep 25;9(9):e108377. doi: 10.1371/journal.pone.0108377. eCollection 2014.
Toxoplasma gondii (T. gondii) is an obligate intracellular protozoan parasite that infects a variety of mammals, including humans. An effective vaccine for this parasite is therefore needed. In this study, RH strain T. gondii rhoptry protein 17 was expressed in bacteria as a fusion with glutathione S-transferase (GST) and the recombinant proteins (rTgROP17) were purified via GST-affinity chromatography. BALB/c mice were nasally immunised with rTgROP17, and induction of immune responses and protection against chronic and lethal T. gondii infections were investigated. The results revealed that mice immunised with rTgROP17 produced high levels of specific anti-rTgROP17 IgGs and a mixed IgG1/IgG2a response of IgG2a predominance. The systemic immune response was associated with increased production of Th1 (IFN-γand IL-2) and Th2 (IL-4) cytokines, and enhanced lymphoproliferation (stimulation index, SI) in the mice immunised with rTgROP17. Strong mucosal immune responses with increased secretion of TgROP17-specific secretory IgA (SIgA) in nasal, vaginal and intestinal washes were also observed in these mice. The vaccinated mice displayed apparent protection against chronic RH strain infection as evidenced by their lower liver and brain parasite burdens (59.17% and 49.08%, respectively) than those of the controls. The vaccinated mice also exhibited significant protection against lethal infection of the virulent RH strain (survival increased by 50%) compared to the controls. Our data demonstrate that rTgROP17 can trigger strong systemic and mucosal immune responses against T. gondii and that ROP17 is a promising candidate vaccine for toxoplasmosis.
刚地弓形虫是一种专性细胞内原生动物寄生虫,可感染包括人类在内的多种哺乳动物。因此,需要一种针对这种寄生虫的有效疫苗。在本研究中,刚地弓形虫RH株棒状体蛋白17在细菌中作为与谷胱甘肽S-转移酶(GST)的融合蛋白表达,重组蛋白(rTgROP17)通过GST亲和层析纯化。用rTgROP17对BALB/c小鼠进行鼻腔免疫,并研究免疫反应的诱导以及对慢性和致死性刚地弓形虫感染的保护作用。结果显示,用rTgROP17免疫的小鼠产生了高水平的特异性抗rTgROP17 IgG,且IgG2a占优势的IgG1/IgG2a混合反应。全身免疫反应与Th1(IFN-γ和IL-2)和Th2(IL-4)细胞因子产生增加以及用rTgROP17免疫的小鼠中淋巴细胞增殖增强(刺激指数,SI)相关。在这些小鼠中还观察到强烈的黏膜免疫反应,鼻腔、阴道和肠道冲洗液中TgROP17特异性分泌型IgA(SIgA)分泌增加。接种疫苗的小鼠对慢性RH株感染表现出明显的保护作用,其肝脏和脑部寄生虫负荷分别比对照组低59.17%和49.08%。与对照组相比,接种疫苗的小鼠对强毒株RH株的致死性感染也表现出显著的保护作用(存活率提高了50%)。我们的数据表明,rTgROP17可引发针对刚地弓形虫的强烈全身和黏膜免疫反应,并且ROP17是弓形虫病的一个有前景的候选疫苗。