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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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Evolution of a functionally intact but antigenically distinct DENV fusion loop.

Rita M Meganck1, Deanna Zhu2, Stephanie Dong2

  • 1Department of Molecular Microbiology and Immunology, Saint Louis University, Saint Louis, United States.

Elife
|September 19, 2023
PubMed
Summary

Researchers engineered a mutated dengue virus (DENV) variant to evade antibody-dependent enhancement, a major challenge for DENV vaccines. This modified DENV may lead to safer vaccines for individuals without prior DENV exposure.

Keywords:
dengue virusevolutionfusion loopinfectious diseasemicrobiologysaturation mutagenesisserumvaccineviruses

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Area of Science:

  • Virology
  • Immunology
  • Vaccinology

Background:

  • Antibody-dependent enhancement (ADE) complicates dengue virus (DENV) vaccine development and is linked to severe secondary DENV infections.
  • ADE is associated with antibodies targeting the conserved fusion loop (FL) motif on the DENV envelope protein, crucial for viral entry.

Purpose of the Study:

  • To engineer a DENV variant with a mutated FL that evades FL-targeting antibodies.
  • To assess the neutralization sensitivity of engineered DENV variants to different antibody types.

Main Methods:

  • Utilized saturation mutagenesis and directed evolution to create a DENV variant with a mutated FL (D2-FL).
  • Combined FL mutations with a modified prM cleavage site to generate a mature DENV variant (D2-FLM).
  • Evaluated neutralization titers using serum from DENV-infected non-human primates (NHP).

Main Results:

  • Engineered DENV variants (D2-FL and D2-FLM) were not neutralized by FL-targeting antibodies.
  • D2-FLM evaded both prM- and FL-targeting antibodies but remained sensitive to other antibodies.
  • Heterotypic (DENV4) NHP serum showed reduced neutralization of D2-FL and D2-FLM compared to wildtype DENV2, while homotypic (DENV2) NHP serum showed similar neutralization.

Conclusions:

  • D2-FL and D2-FLM are valuable tools for characterizing cross-reactive (CR) antibody subtypes.
  • These engineered DENV variants represent a promising platform for developing safer live-attenuated DENV vaccines, particularly for naive individuals and children.