远古诺卡氏菌 IFM10152 的分泌蛋白 NFA47630 可诱导小鼠产生免疫保护作用。

IF 2.4 Q3 INFECTIOUS DISEASES Tropical Diseases, Travel Medicine and Vaccines Pub Date : 2024-10-15 DOI:10.1186/s40794-024-00229-w
Lichao Han, Xingzhao Ji, Shihong Fan, Jirao Shen, Bin Liang, Zhenjun Li
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引用次数: 0

摘要

目的:诺卡氏菌正在成为全球常见且易被忽视的医疗保健和职业相关感染的病因,但目前还没有用于预防诺卡氏菌的人类疫苗。本研究旨在评估 NFA47630 蛋白的免疫保护作用,NFA47630 蛋白是一种在 N. farcinica IFM10152 上清液中含量丰富的分泌蛋白:方法:通过 PCR 和生物信息学分析了 NFA47630 的保守性和特征。然后克隆、表达和纯化重组 NFA47630 蛋白,进一步进行抗原性分析。随后,通过检测 MAPK 信号通路的磷酸化状态和细胞因子水平来评估其激活先天性免疫的能力。最后,对rNFA47630免疫小鼠的保护作用进行了评估。rNFA47630蛋白在0.2 mM IPTG、28 °C的最佳条件下表达,能被抗N. farcinica和抗N. cyriacigeorgica血清识别,但不能被抗N. asteroids、抗N. brasiliensis、抗N. nova和抗牛分枝杆菌血清识别。它能上调 ERK、JNK、P38 的磷酸化状态以及 TNF-α、IL-10、IL-12 和 IFN-γ 等细胞因子的水平。此外,与PBS免疫小鼠相比,rNFA47630蛋白免疫小鼠表现出更高的抗体滴度、更强的细菌清除能力、更轻的器官感染以及更高的存活率:我们的数据表明,NFA47630是一种潜在的候选疫苗,可用于防御远华蓟马感染。
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Secreted protein NFA47630 from Nocardia farcinica IFM10152 induces immunoprotective effects in mice.

Purpose: Nocardia is emerging as a common and easily neglected cause of both healthcare- and occupation-associated infections worldwide, however, human vaccines for Nocardia prevention are not yet available. In this study, we aimed to evaluate the immunoprotective effect of the NFA47630 protein, a secreted protein abundant in the N. farcinica IFM10152 supernatant.

Methods: Conservation and characteristics of nfa47630 were analyzed by PCR and bioinformatics. Then recombinant NFA47630 protein was cloned, expressed and purified for further antigenicity analysis. Subsequently, the ability to activate innate immunity was evaluated by examining the phosphorylation status of the MAPK signaling pathway and cytokine levels. Finally, the protective effect was evaluated on rNFA47630-immunized mice.

Results: nfa47630 was conserved in N. farcinica strains with good antigenicity. The rNFA47630 protein was expressed under the optimal conditions of 0.2 mM IPTG, 28 °C, and it can be recognized by anti-N. farcinica and anti-N. cyriacigeorgica sera, but not anti-N. asteroids, anti-N. brasiliensis, anti-N. nova and anti-Mycobacterium bovis sera. It can upregulate the phosphorylation status of ERK, JNK, P38 and the cytokine levels of TNF-α, IL-10, IL-12, and IFN-γ. In addition, mice immunized with rNFA47630 protein exhibited higher antibody titers, greater bacterial clearance ability, milder organ infection, and higher survival rates than PBS-immunized mice.

Conclusions: Our data demonstrate that NFA47630 is a potential vaccine candidate for defending against N. farcinica infection.

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来源期刊
CiteScore
5.20
自引率
0.00%
发文量
25
审稿时长
17 weeks
期刊介绍: Tropical Diseases, Travel Medicine and Vaccines is an open access journal that considers basic, translational and applied research, as well as reviews and commentary, related to the prevention and management of healthcare and diseases in international travelers. Given the changes in demographic trends of travelers globally, as well as the epidemiological transitions which many countries are experiencing, the journal considers non-infectious problems including chronic disease among target populations of interest as well as infectious diseases.
期刊最新文献
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