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  • Development of an adjuvanted nanoparticle vaccine against influenza virus, an in vitro study.

Development of an adjuvanted nanoparticle vaccine against influenza virus, an in vitro study.

PloS one (2020-08-08)
Kamonthip Rungrojcharoenkit, Panya Sunintaboon, Damon Ellison, Louis Macareo, Panuwat Midoeng, Preamrudee Chaisuwirat, Stefan Fernandez, Sukathida Ubol
要旨

Influenza is an infectious respiratory illness caused by influenza viruses. Despite yearly updates, the efficacy of influenza vaccines is significantly curtailed by the virus antigenic drift and antigenic shift. These constant changes to the influenza virus make-up also challenge the development of a universal flu vaccine, which requires conserved antigenic regions shared by influenza viruses of different subtypes. We propose that it is possible to bypass these challenges by the development of an influenza vaccine based on conserved proteins delivered in an adjuvanted nanoparticle system. In this study, we generated influenza nanoparticle constructs using trimethyl chitosan nanoparticles (TMC nPs) as the carrier of recombinant influenza hemagglutinin subunit 2 (HA2) and nucleoprotein (NP). The purified HA2 and NP recombinant proteins were encapsulated into TMC nPs to form HA2-TMC nPs and NP-TMC nPs, respectively. Primary human intranasal epithelium cells (HNEpCs) were used as an in vitro model to measure immunity responses. HA2-TMC nPs, NP-TMC nPs, and HA2-NP-TMC nPs (influenza nanoparticle constructs) showed no toxicity in HNEpCs. The loading efficiency of HA2 and NP into the TMC nPs was 97.9% and 98.5%, respectively. HA2-TMC nPs and NP-TMC nPs more efficiently delivered HA2 and NP proteins to HNEpCs than soluble HA2 and NP proteins alone. The induction of various cytokines and chemokines was more evident in influenza nanoparticle construct-treated HNEpCs than in soluble protein-treated HNEpCs. In addition, soluble factors secreted by influenza nanoparticle construct-treated HNEpCs significantly induced MoDCs maturation markers (CD80, CD83, CD86 and HLA-DR), as compared to soluble factors secreted by protein-treated HNEpCs. HNEpCs treated with the influenza nanoparticle constructs significantly reduced influenza virus replication in an in vitro challenge assay. The results indicate that TMC nPs can be used as influenza vaccine adjuvants and carriers capable of delivering HA2 and NP proteins to HNEpCs.

材料
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製品内容

Sigma-Aldrich
meso-テトラフェニルポルフィリン, BioReagent, suitable for fluorescence, ≥99.0% (HPLC)
Sigma-Aldrich
スパウチン-1, ≥98% (HPLC)
Sigma-Aldrich
Anti-Influenza A Antibody, nucleoprotein, clone A1, FITC-conjugated, clone A1, Chemicon®, from mouse

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