Construction of engineered probiotic that adhere and display nanobody to neutralize porcine reproductive and respiratory syndrome virus

IF 2.3 3区生物学 Q3 MICROBIOLOGY

Archives of Microbiology Pub Date : 2024-11-14 DOI:10.1007/s00203-024-04198-8

Li Zhou, Hanlin Zhou, Panying Wang, Hang Xu, Jiayi Wu, Yuanzhuo Zhou, Jiaying Feng, Weiyi Zheng

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引用次数: 0

Abstract

Pathogenic blue ear disease caused by porcine reproductive and respiratory syndrome virus (PRRSV) bring severe loss to breeding industry due to high infectivity and mortality. L. plantarum serves as the probiotic host strain, known for its beneficial properties in the gut microbiota. E. coli is used as a cloning host for the initial genetic engineering steps, facilitating the construction and amplification of the desired genetic constructs. In this study, using synthetic biology technology, we constructed engineered probiotics which could adhere and display nanobody on the surface to neutralize virus. Firstly, we screen an optimal nanobody to effectively bind with PRRSV by building library, expression and purification. Then, the integration of adhesion protein and nanobody into the genome of probiotics significantly improved its adhesion to IPEC-J2 cells. In addition, this engineered probiotic is almost non-toxic to cells with good safety, which can be used as a daily probiotics to prevent virus fecal transmission. Our study proposed this novel construction strategy of engineering probiotics with both adhesion and neutralization effects, which provided a new therapeutic view for intestinal virus clearance.

Graphical Abstract

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构建可粘附和显示纳米抗体的工程益生菌，以中和猪繁殖与呼吸综合征病毒。

猪繁殖与呼吸综合征病毒（PRRSV）引起的致病性蓝耳病传染性强、死亡率高，给养殖业带来严重损失。植物乳杆菌作为益生菌宿主菌株，因其在肠道微生物群中的有益特性而闻名。大肠杆菌被用作基因工程初始步骤的克隆宿主，便于构建和扩增所需的基因构建体。在本研究中，我们利用合成生物学技术构建了工程益生菌，这种益生菌可以粘附并在表面显示纳米抗体，从而中和病毒。首先，我们通过建库、表达和纯化筛选出能与 PRRSV 有效结合的最佳纳米抗体。然后，将粘附蛋白和纳米抗体整合到益生菌基因组中，大大提高了益生菌对 IPEC-J2 细胞的粘附力。此外，这种工程益生菌对细胞几乎无毒，具有良好的安全性，可作为日常益生菌预防病毒粪便传播。我们的研究提出了这种既有粘附作用又有中和作用的新型工程益生菌构建策略，为清除肠道病毒提供了新的治疗思路。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Archives of Microbiology 生物-微生物学

CiteScore

4.90

自引率

3.60%

发文量

601

审稿时长

3 months

期刊介绍： Research papers must make a significant and original contribution to microbiology and be of interest to a broad readership. The results of any experimental approach that meets these objectives are welcome, particularly biochemical, molecular genetic, physiological, and/or physical investigations into microbial cells and their interactions with their environments, including their eukaryotic hosts. Mini-reviews in areas of special topical interest and papers on medical microbiology, ecology and systematics, including description of novel taxa, are also published. Theoretical papers and those that report on the analysis or ''mining'' of data are acceptable in principle if new information, interpretations, or hypotheses emerge.