IF 3.9 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Applied Microbiology and Biotechnology Pub Date : 2025-03-03 DOI:10.1007/s00253-025-13436-y
Isabelle Loop, Yuan-Dun Ke, Wei-June Chen, Kun-Hsien Tsai, Wei-Li Hsu, Yi-Chin Fan
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引用次数: 0

摘要

蜱传严重发热伴血小板减少综合征(SFTS)是一种由SFTS病毒(SFTSV)引起的新兴人畜共患病。基于该病毒核壳蛋白和部分糖蛋白的血清学检测方法已被用于检测人和动物的 SFTSV 感染。然而,完整的 SFTSV 糖蛋白(Gn/Gc)能否诱导病毒样颗粒(VLPs)的组装,从而用于血清学监测和疫苗生产,目前仍不清楚。在本研究中,我们使用单一质粒编码显性基因 B 型病毒的完整糖蛋白序列,成功表达并分泌了 SFTSV Gn/Gc 抗原。用上述质粒转染 HEK293T 和 COS-1 细胞;在 32 °C 而不是 37 °C 下培养这些细胞,抗原回收率分别提高了 4.0 倍和 3.3 倍。32 ℃下分泌的Gn/Gc抗原保留了与病毒表位相似的表位;这些表位可被SFTS人源单克隆抗体识别。蔗糖密度梯度离心,然后用透射电子显微镜观察,证实形成了直径约为 100 nm 的 VLPs。总之,我们的研究结果凸显了 SFTSV VLPs 在血清学监测和疫苗开发方面的潜力。
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Low-temperature enhances production of severe fever with thrombocytopenia syndrome virus virus-like particles

Tick-borne severe fever with thrombocytopenia syndrome (SFTS) is an emerging zoonotic disease caused by the SFTS virus (SFTSV). Serological assays based on the nucleocapsid protein and partial glycoprotein of this virus have been used for detecting SFTSV infections in humans and animals. However, whether the complete SFTSV glycoprotein (Gn/Gc) can induce the assembly of virus-like particles (VLPs) which can be used for serological surveillance and vaccine production remains unclear. In this study, we successfully expressed and secreted SFTSV Gn/Gc antigens by using a single plasmid encoding the complete glycoprotein sequence of the dominant genotype B virus. HEK293T and COS-1 cells were transfected with the aforementioned plasmid; cultivating these cells at 32 °C, instead of 37 °C, led to 4.0- and 3.3-fold higher antigen recovery, respectively. The secreted Gn/Gc antigens at 32 °C retained epitopes resembling those of the virion; these epitopes were recognized by a SFTS human–derived monoclonal antibody. Sucrose density gradient centrifugation, followed by transmission electron microscopy, confirmed the formation of VLPs with a diameter of approximately 100 nm. Overall, our findings highlight the potential of SFTSV VLPs for serological surveillance and vaccine development.

• Cultivating transfected cells at 32 °C boosts SFTSV glycoprotein production.

• Complete SFTSV glycoprotein expression facilitates virus-like particle assembly.

• The assembly does not require any other viral proteins or RNA.

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来源期刊
Applied Microbiology and Biotechnology
Applied Microbiology and Biotechnology 工程技术-生物工程与应用微生物
CiteScore
10.00
自引率
4.00%
发文量
535
审稿时长
2 months
期刊介绍: Applied Microbiology and Biotechnology focusses on prokaryotic or eukaryotic cells, relevant enzymes and proteins; applied genetics and molecular biotechnology; genomics and proteomics; applied microbial and cell physiology; environmental biotechnology; process and products and more. The journal welcomes full-length papers and mini-reviews of new and emerging products, processes and technologies.
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