Development and optimisation of Influenza C and Influenza D pseudotyped viruses

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

doi:10.1016/j.jviromet.2025.115243

ImmunologyJournal of Virological MethodsAug 23, 2025

Peer-reviewed journal article

Development and optimisation of Influenza C and Influenza D pseudotyped viruses

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

PDF DOI Preprint

Post to X

Threads on X Post to Bluesky Threads on Bluesky

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.