Monitoring Neutralization Property Change of Evolving Hantaan and Seoul Viruses with a Novel Pseudovirus-Based Assay |
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Authors: | Ning Tingting Wang Ling Liu Shuo Ma Jian Nie Jianhui Huang Weijin Li Xuguang Li Yuhua Wang Youchun |
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Institution: | 1.Division of HIV/AIDS and Sexually Transmitted Virus Vaccines, National Institutes for Food and Drug Control(NIFDC), Beijing 102629, China2.Division of Arboviruses Vaccine, National Institutes for Food and Drug Control(NIFDC), Beijing 102629, China3.Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, ON K2J 4X9, Canada |
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Abstract: | 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(VSVDG*-HTNVG or VSVDG*-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 VSVDG*-HTNVG and 0.82 for VSVDG*-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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Keywords: | Hemorrhagic fever with renal syndrome (HFRS) Hantaan virus (HTNV) Seoul virus (SEOV) Pseudovirus-based neutralization assay (PBNA) Amino acid substitution Vaccine |
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