The Hantaan virus (HTNV) and Seoul virus (SEOV) mutants have accumulated over time. It is important to determine whether their neutralizing epitopes have evolved, thereby making the current vaccine powerless. However, it is impossible to determine by using traditional plaque reduction neutralization test (PRNT), because it requires large numbers of live mutant strains. Pseudovirus-based neutralization assays (PBNA) were developed by employing vesicular stomatitis virus (VSV) backbone incorporated with HTNV or SEOV glycoproteins (VSVΔG*-HTNVG or VSVΔG*-SEOVG). 56 and 51 single amino acid substitutions of glycoprotein (GP) in HTNV and SEOV were selected and introduced into the reference plasmid. Then the mutant pseudoviruses were generated and tested by PBNA. The PBNA results were highly correlated with PRNT ones with R2 being 0.91 for VSVΔG*-HTNVG and 0.82 for VSVΔG*-SEOVG. 53 HTNV mutant pseudoviruses and 46 SEOV mutants were successfully generated. Importantly, by using PBNA, we found that HTNV or SEOV immunized antisera could neutralize all the corresponding 53 HTNV mutants or the 46 SEOV mutants respectively. The novel PBNA enables us to closely monitor the effectiveness of vaccines against large numbers of evolving HTNV and SEOV. And the current vaccine remains to be effective for the naturally occurring mutants.
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We would like to thank M. A. Whitt for providing the VSVΔG* pseudotype virus bearing VSV GP (VSVΔG*-G), Arboviruses Vaccine Division for HTNV strain 76-118 and Seoul virus UR, Changchun Institute of Biological Products Co., Ltd. (Changchun, China), Aimei Weixin Biopharmaceutical (Zhejiang) Co., Ltd. (Zhejiang, China), and Royal (Wuxi) Bio-pharmaceutical Co., Ltd. (Jiangsu, China) for their HFRS bivalent vaccines, rabbit antisera against HTNV and rabbit antisera against SEOV. The study was supported by the National Science and Technology Major Projects of Drug Discovery [Grant Number 2018ZX09101-001]
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The authors declare that they have no conflict of interest.
Animal and Human Rights Statement
The whole study was approved by the Institutional Animal Care and Use Committee of National Institutes for Food and Drug Control, China. All institutional and national guidelines for the care and use of laboratory animals were followed.
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Ning, T., Wang, L., Liu, S. et al. Monitoring Neutralization Property Change of Evolving Hantaan and Seoul Viruses with a Novel Pseudovirus-Based Assay. Virol. Sin. 36, 104–112 (2021). https://doi.org/10.1007/s12250-020-00237-y
- Hemorrhagic fever with renal syndrome (HFRS)
- Hantaan virus (HTNV)
- Seoul virus (SEOV)
- Pseudovirus-based neutralization assay (PBNA)
- Amino acid substitution