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弓形虫 RAP 结构域结合蛋白(rTgRA15)的免疫刺激作用。

Immune Stimulation of RAP domain binding protein (rTgRA15) from Toxoplasma gondii.

机构信息

a Institute for Research in Molecular Medicine (INFORMM) , Universiti Sains Malaysia , Minden , Penang , Malaysia.

出版信息

Pathog Glob Health. 2018 Oct;112(7):387-394. doi: 10.1080/20477724.2018.1536854. Epub 2018 Oct 17.

Abstract

Toxoplasmosis, a parasitic disease in human and animals, is caused by Toxoplasma gondii. Our previous study has led to the discovery of a novel RAP domain binding protein antigen (TgRA15), an apparent in-vivo induced antigen recognised by antibodies in acutely infected individuals. This study is aimed to evaluate the humoral response and cytokine release elicited by recombinant TgRA15 protein in C57BL/6 mice, demonstrating its potential as a candidate vaccine for Toxoplasma gondii infection. In this study, the recombinant TgRA15 protein was expressed in Escherichia coli, purified and refolded into soluble form. C57BL/6 mice were immunised intradermally with the antigen and CASAC (Combined Adjuvant for Synergistic Activation of Cellular immunity). Antigen-specific humoral and cell-mediated responses were evaluated using Western blot and ELISA. The total IgG, IgG and IgG antibodies specific to the antigen were significantly increased in treatment group compare to control group. A higher level of interferon gamma (IFN-γ) secretion was demonstrated in the mice group receiving booster doses of rTgRA15 protein, suggesting a potential Th1-mediated response. In conclusion, the rTgRA15 protein has the potential to generate specific antibody response and elicit cellular response, thus potentially serve as a vaccine candidate against T. gondii infection.

摘要

弓形虫病是一种人和动物的寄生虫病,由刚地弓形虫引起。我们之前的研究发现了一种新的 RAP 结构域结合蛋白抗原(TgRA15),这是一种明显的体内诱导抗原,被急性感染个体的抗体识别。本研究旨在评估重组 TgRA15 蛋白在 C57BL/6 小鼠中引起的体液反应和细胞因子释放,证明其作为刚地弓形虫感染候选疫苗的潜力。在这项研究中,重组 TgRA15 蛋白在大肠杆菌中表达,纯化并复性为可溶性形式。用抗原和 CASAC(协同激活细胞免疫的联合佐剂)皮内免疫 C57BL/6 小鼠。使用 Western blot 和 ELISA 评估抗原特异性体液和细胞介导的反应。与对照组相比,治疗组中针对抗原的总 IgG、IgG 和 IgG 抗体特异性显著增加。在接受 rTgRA15 蛋白增强剂量的小鼠组中,干扰素γ(IFN-γ)的分泌水平更高,表明存在潜在的 Th1 介导的反应。总之,rTgRA15 蛋白具有产生特异性抗体反应和引发细胞反应的潜力,因此可能成为针对刚地弓形虫感染的候选疫苗。

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