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弓形虫 eIF4A 表达 DNA 疫苗诱导的保护性免疫对小鼠急性弓形虫病的作用。

Protective immunity induced by a DNA vaccine expressing eIF4A of Toxoplasma gondii against acute toxoplasmosis in mice.

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 730046, PR China.

出版信息

Vaccine. 2013 Mar 25;31(13):1734-9. doi: 10.1016/j.vaccine.2013.01.027. Epub 2013 Jan 29.

Abstract

Toxoplasma gondii is an obligate intracellular protozoan parasite infecting humans, mammals and birds. Eukaryotic translation initiation factor (eIF4A) is a newly identified protein associated with tachyzoite virulence. To evaluate the protective efficacy of T. gondii eIF4A, a DNA vaccine (pVAX-eIF4A) encoding T. gondii eIF4A (Tg-eIF4A) gene was constructed. The expression ability of this recombinant DNA plasmid was examined in Marc145 cells by IFA. Then, Kunming mice were intramuscularly immunized with pVAX-eIF4A and followed by challenge infection with the highly virulent T. gondii RH strain. The results showed that vaccination with pVAX-eIF4A elicited specific humoral responses, with high IgG antibody titers and specific lymphocyte proliferative responses. The cellular immune response was associated with significant production of IFN-γ, IL-2 in Kunming mice, and a mixed IgG1/IgG2a response with predominance of IgG2a production, indicating that a Th1 type response was elicited after immunization with pVAX-eIF4A. In addition, the increase of the percentage of CD8+ T cells in lymphoid in mice suggested the activation of MHC class I restricted antigen presentation pathways. After lethal challenge, the mice vaccinated with the pVAX-eIF4A showed a significantly prolonged survival time (23.0±5.5 days) compared with control mice which died within 7 days of challenge (P<0.05). These results demonstrate that pVAX-eIF4A could elicit strong humoral, Th1-type cellular immune responses and increase survival time of immunized mice, suggesting that eIF4A is a promising vaccine candidate against acute T. gondii infection in mice.

摘要

刚地弓形虫是一种专性细胞内寄生的原虫,感染人类、哺乳动物和鸟类。真核翻译起始因子(eIF4A)是一种新鉴定的与速殖子毒力相关的蛋白。为评估弓形虫 eIF4A 的保护效力,构建了编码弓形虫 eIF4A(Tg-eIF4A)基因的 DNA 疫苗(pVAX-eIF4A)。通过 IFA 检测该重组 DNA 质粒在 Marc145 细胞中的表达能力。然后,昆明小鼠肌肉内免疫接种 pVAX-eIF4A,并用高毒力弓形虫 RH 株进行攻毒感染。结果表明,pVAX-eIF4A 疫苗接种可诱导特异性体液反应,产生高 IgG 抗体滴度和特异性淋巴细胞增殖反应。细胞免疫反应与 IFN-γ、IL-2 在昆明小鼠中的显著产生有关,以及 IgG1/IgG2a 的混合应答,以 IgG2a 产生为主,表明 pVAX-eIF4A 免疫后诱导了 Th1 型反应。此外,小鼠淋巴样中 CD8+T 细胞比例的增加提示 MHC Ⅰ类限制的抗原呈递途径被激活。在致死性攻毒后,与对照组小鼠在攻毒后 7 天内死亡相比,接种 pVAX-eIF4A 的小鼠存活时间明显延长(23.0±5.5 天)(P<0.05)。这些结果表明,pVAX-eIF4A 可诱导强烈的体液、Th1 型细胞免疫反应,并增加免疫接种小鼠的存活时间,表明 eIF4A 是一种有前途的抗急性弓形虫感染的候选疫苗。

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