Journal
ARCHIVES OF VIROLOGY
Volume 168, Issue 2, Pages -Publisher
SPRINGER WIEN
DOI: 10.1007/s00705-022-05697-z
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The intron-based stabilization approach is useful for stable flavivirus infectious clone construction. A highly attenuated Japanese encephalitis live vaccine strain (SA(14)-14-2) was used as a backbone vector to develop recombinant vaccines. However, a rescued virus strain (rHV-DJEV) with increased neurovirulence was accidentally acquired during the experiments, indicating the need for caution in using the intron-based stabilization approach in vaccine development.
The intron-based stabilization approach is a very useful strategy for construction of stable flavivirus infectious clones. SA(14)-14-2 is a highly attenuated Japanese encephalitis (JE) live vaccine strain that has been widely used in China since 1989. To develop safe and effective recombinant vaccines with SA(14)-14-2 as a backbone vector, we constructed the DNA-based infectious clone pCMW-JEV of SA(14)-14-2 using the intron-based stabilization approach and acquired the rescued virus rDJEV, which retained the biological properties of the parental virus. Unexpectedly, a rescued virus strain with altered virulence, designated rHV-DJEV, was accidentally acquired in one of the transfection experiments. rHV-DJEV showed up to 10(5)-fold increased neurovirulence compared with the SA(14)-14-2 parental strain. Genome sequencing showed that the inserted introns were still present in the genome of rHV-DJEV. Therefore, we think that the intron-based stabilization approach should be used with caution in vaccine development and direct iDNA immunization.
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