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弓形虫棒状体蛋白13质粒DNA疫苗对小鼠的保护效力

Protective efficacy of a Toxoplasma gondii rhoptry protein 13 plasmid DNA vaccine in mice.

作者信息

Wang Pei-Yuan, Yuan Zi-Guo, Petersen Eskild, Li Jie, Zhang Xiu-Xiang, Li Xiu-Zhen, Li Hao-Xin, Lv Zhi-Cheng, Cheng Tian, Ren Di, Yang Gui-Lian, Lin Rui-Qing, Zhu Xing-Quan

机构信息

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, People's Republic of China.

出版信息

Clin Vaccine Immunol. 2012 Dec;19(12):1916-20. doi: 10.1128/CVI.00397-12. Epub 2012 Sep 26.

Abstract

Toxoplasma gondii is an obligate intracellular parasite infecting humans and other warm-blooded animals, resulting in serious public health problems and economic losses worldwide. Rhoptries are involved in T. gondii invasion and host cell interaction and have been implicated as important virulence factors. In the present study, a DNA vaccine expressing rhoptry protein 13 (ROP13) of T. gondii inserted into eukaryotic expression vector pVAX I was constructed, and the immune protection it induced in Kunming mice was evaluated. Kunming mice were immunized intramuscularly with pVAX-ROP13 and/or with interleukin-18 (IL-18). Then, we evaluated the immune response using a lymphoproliferative assay, cytokine and antibody measurements, and the survival times of mice challenged with the virulent T. gondii RH strain (type I) and the cyst-forming PRU strain (type II). The results showed that pVAX-ROP13 alone or with pVAX/IL-18 induced a high level of specific anti-T. gondii antibodies and specific lymphocyte proliferative responses. Coinjection of pVAX/IL-18 significantly increased the production of gamma interferon (IFN-γ), IL-2, IL-4, and IL-10. Further, challenge experiments showed that coimmunization of pVAX-ROP13 with pVAX/IL-18 significantly (P < 0.05) increased survival time (32.3 ± 2.7 days) compared with pVAX-ROP13 alone (24.9 ± 2.3 days). Immunized mice challenged with T. gondii cysts (strain PRU) had a significant reduction in the number of brain cysts, suggesting that ROP13 could trigger a strong humoral and cellular response against T. gondii cyst infection and that it is a potential vaccine candidate against toxoplasmosis, which provided the foundation for further development of effective vaccines against T. gondii.

摘要

刚地弓形虫是一种专性细胞内寄生虫,可感染人类和其他温血动物,在全球范围内导致严重的公共卫生问题和经济损失。棒状体参与刚地弓形虫的入侵和宿主细胞相互作用,并被认为是重要的毒力因子。在本研究中,构建了一种将刚地弓形虫棒状体蛋白13(ROP13)插入真核表达载体pVAX I的DNA疫苗,并评估了其在昆明小鼠中诱导的免疫保护作用。用pVAX-ROP13和/或白细胞介素-18(IL-18)对昆明小鼠进行肌肉注射免疫。然后,我们使用淋巴细胞增殖试验、细胞因子和抗体检测以及用强毒株刚地弓形虫RH株(I型)和形成包囊的PRU株(II型)攻击小鼠后的存活时间来评估免疫反应。结果表明,单独的pVAX-ROP13或与pVAX/IL-18联合使用可诱导高水平的特异性抗刚地弓形虫抗体和特异性淋巴细胞增殖反应。pVAX/IL-18的共同注射显著增加了γ干扰素(IFN-γ)、IL-2、IL-4和IL-10的产生。此外,攻毒实验表明,与单独使用pVAX-ROP13(24.9±2.3天)相比,pVAX-ROP13与pVAX/IL-18共同免疫显著(P<0.05)延长了存活时间(32.3±2.7天)。用刚地弓形虫包囊(PRU株)攻击的免疫小鼠脑内包囊数量显著减少,这表明ROP13可引发针对刚地弓形虫包囊感染的强烈体液和细胞反应,并且它是一种抗弓形虫病的潜在疫苗候选物,这为进一步开发有效的抗刚地弓形虫疫苗奠定了基础。

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