Development and optimisation of influenza C and influenza D pseudotyped viruses

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2025-08-23 DOI:10.1016/j.jviromet.2025.115243

Maria Giovanna Marotta , Martin Mayora Neto , Janet Daly , Meshach Maina , Pauline van Diemen , Helen Everett , Maria Stella Lucente , Michele Camero , Emanuele Montomoli , Claudia Maria Trombetta , Kelly da Costa , Nigel J. Temperton

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引用次数: 0

Abstract

To facilitate the study of influenza C (ICV) and influenza D (IDV) viruses, we generated lentiviral pseudotyped viruses (PVs) expressing the hemagglutinin-esterase fusion (HEF) glycoprotein from ICV (C/Minnesota/33/2015) and IDV (D/Swine/Italy/199724–3/2015, D/Bovine/France/5920/2014, and D/Bovine/Ibaraki/7768/2016). The production of these PVs was optimised using different amount of human airway trypsin-like (HAT) protease to enhance HEF maturation, and the transduction efficiency was evaluated in multiple cell lines. Using these PVs, we established a pseudovirus-based microneutralisation (pMN) assay to measure neutralising antibody responses and adapted an esterase activity assay to evaluate PV. Specific antisera neutralised PVs but failed to inhibit esterase activity. These findings confirm that ICV and IDV PVs provide a scalable, sensitive, and safe tool for antiviral screening, and sero-epidemiological research.

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流感C型和流感D型假病毒的研制和优化

为了促进流感C （ICV）和流感D （IDV）病毒的研究，我们从ICV （C/Minnesota/33/2015）和IDV （D/Swine/Italy/ 199724-3/2015、D/Bovine/France/5920/2014和D/Bovine/Ibaraki/7768/2016）中制备了表达血凝素-酯酶融合（HEF）糖蛋白的慢病毒假型病毒（PVs）。使用不同量的人气道胰蛋白酶样（human airway trypsin-like， HAT）蛋白酶来优化这些pv的产生，以促进HEF的成熟，并在多个细胞系中评估其转导效率。利用这些PV，我们建立了一种基于假病毒的微中和（pMN）试验来测量中和抗体反应，并采用酯酶活性试验来评估PV。特异性抗血清能中和pv，但不能抑制酯酶活性。这些发现证实，ICV和IDV pv为抗病毒筛查和血清流行病学研究提供了一种可扩展、敏感和安全的工具。

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.