Evolution of a Functionally Intact but Antigenically Distinct DENV Fusion Loop

• DOI: 10.7554/elife.87555.1
• Publication Date: 2023-07-12T15:30:56Z
• Authors: Rita M. Meganck; Deanna Zhu; Stephanie Dong; Lisa J. Snoderly-...; Yago R. Dalben; Devina Thiono; Laura J. White; Aravinda M. DeSilva; Ralph S. Baric; Longping V. Tse
• Keywords: Microbiology and Infectious Disease; Vaccines; dengue virus; QH301-705.5; Science; Q; R; saturation mutagenesis; Antibodies, Monoclonal; fusion loop; Cross Reactions; Article; 3. Good health; Culicidae; Engineering; vaccine; evolution; Medicine; Animals; Biology (General); serum

Abstract A hallmark of Dengue virus (DENV) pathogenesis is the potential for antibody-dependent enhancement, which is associated with deadly DENV secondary infection, complicates the identification of correlates of protection, and negatively impacts the safety and efficacy of DENV vaccines. Antibody-dependent enhancement is linked to antibodies targeting the fusion loop (FL) motif of the envelope protein, which is completely conserved in mosquito-borne flaviviruses and required for viral entry and fusion. In the current study, we utilized saturation mutagenesis and directed evolution to engineer a functional variant with a mutated fusion-loop (D2-FL) which is not neutralized by fusion-loop-targeting monoclonal antibodies. The fusion-loop mutations were combined with our previously evolved pre-membrane cleavage site to create a mature version of D2-FL (D2-FLM), which evades both pre-membrane and fusion-loop antibodies but retains sensitivity to other type-specific and quaternary cross-reactive antibodies. Cross-reactive serum from heterotypic (DENV4) infected non-human primates showed lower neutralization titers against D2-FL and D2-FLM than isogenic wildtype DENV2 while similar neutralization titers were observed in serum from homotypic (DENV2) infected non-human primates. We propose D2-FL and D2-FLM as valuable tools to delineate cross-reactive antibody subtypes in serum as well as an exciting platform for safer live attenuated DENV vaccines suitable for naïve individuals and children.