Preparation and verification of highly pathogenic H5N8 subtype avian influenza pseudovirus

HU Minhao; LUO Wanrong; XIA Yidan; YU Junling; ZHOU Xue; FANG Weixi; YANG Sitian; HE Jun

doi:10.19837/j.cnki.ahyf.2024.01.001

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Anhui Journal of Preventive Medicine ›› 2024, Vol. 30 ›› Issue (1) : 1-5. DOI: 10.19837/j.cnki.ahyf.2024.01.001

Special Contributions

Preparation and verification of highly pathogenic H5N8 subtype avian influenza pseudovirus

HU Minhao^1,2,3, LUO Wanrong^1,2, XIA Yidan³, YU Junling^1,2, ZHOU Xue^1,2, FANG Weixi^1,2, YANG Sitian³, HE Jun^1,2,3

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Abstract

Objective In order to study the structure of the highly pathogenic H5N8 subtype (A/Astrakhan/3212/2020) avian influenza virus,the corresponding pseudovirus was prepared and its infective ability at the cellular level,as well as the neutralizing ability of serum antibodies,were preliminarily verified. Methods By analyzing the amino acid sequences of the H5N8 subtype (A/Astrakhan/3212/2020) virus hemagglutinin (HA) and neuraminidase (NA),the target HA and NA sequences were optimized and the corresponding plasmid was extracted.Moreover,H5N8 subtype pseudovirus was prepared by a lentivirus packaging system.The fabricated pseudovirus was verified by luciferase readings,hemagglutination inhibition tests,transmission electron microscopy,and serum neutralization tests after infection with 293T-ST6GAL1 cells containing α-2,6 sialic acid receptors. Results H5N8 subtype pseudovirus was successfully prepared,and its infectivity to 293T-ST6GAL1 cells was 10³ TCID50/mL.After being 50 times-concentrated,the hemagglutination activity of the pseudovirus increased from 16 to 128.Typical influenza virus particles could be observed by transmission electron microscopy.The neutralization test results showed that the neutralization activity of 3 H5N8 positive sera was significantly higher than that of other positive sera and negative sera. Conclusion This study successfully prepared the highly pathgenic H5N8 subtype avian influenza pseudovirus,which provides an important experimental platform for future vaccine development and screening.

Key words

H5N8 subtype avian influenza virus / Pseudovirus preparation / Hemagglutinin / Neuraminidase / Neutralization test

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HU Minhao, LUO Wanrong, XIA Yidan, YU Junling, ZHOU Xue, FANG Weixi, YANG Sitian, HE Jun. Preparation and verification of highly pathogenic H5N8 subtype avian influenza pseudovirus[J]. Anhui Journal of Preventive Medicine. 2024, 30(1): 1-5 https://doi.org/10.19837/j.cnki.ahyf.2024.01.001

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