Journal
WORLD JOURNAL OF PEDIATRICS
Volume 15, Issue 5, Pages 499-505Publisher
ZHEJIANG UNIV SCH MEDICINE
DOI: 10.1007/s12519-019-00286-8
Keywords
Mumps virus; Reverse genetics; Plasmid; Safety; Immunogenicity
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Funding
- Natural Science Foundation for Young Scholars of Zhejiang Province [LQ19H100005]
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Background Mumps is a common type of respiratory infectious disease caused by mumps virus (MuV), and can be effectively prevented by vaccination. In this study, a reverse genetic system of MuV that can facilitate the rational design of safer, more efficient mumps vaccine candidates is established. Methods MuV-S79 cDNA clone was assembled into a full-length plasmid by means of the GeneArt (TM) High-Order Genetic Assembly System, and was rescued via reverse genetic technology. RT-PCR, sequencing, and immunofluorescence assays were used for rMuV-S79 authentication. Viral replication kinetics and in vivo experimental models were used to evaluate the replication, safety, and immunogenicity of rMuV-S79. Results A full-length cDNA clone of MuV-S79 in the assembly process was generated by a novel plasmid assemble strategy, and a robust reverse genetic system of MuV-S79 was successfully established. The established rMuV-S79 strain could reach a high virus titer in vitro. The average viral titer of rMuV-S79 in the lung tissues was 2.68 +/- 0.14 log(10)PFU/g lung tissue, and rMuV-S79 group did not induce inflammation in the lung tissues in cotton rats. Neutralizing antibody titers induced by rMuV-S79 were high, long-lasting and could provide complete protection against MuV wild strain challenge. Conclusion We have established a robust reverse genetic system of MuV-S79 which can facilitate the optimization of mumps vaccines. rMuV-S79 rescued could reach a high virus titer and the safety was proven in vivo. It could also provide complete protection against MuV wild strain challenge.
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