Evolution of a functionally intact but antigenically distinct DENV fusion loop
- Department of Molecular Microbiology and Immunology, Saint Louis University, United States
- Department of Epidemiology, University of North Carolina at Chapel Hill, United States
- Department of Microbiology, University of North Carolina at Chapel Hill, United States
Figures
Figure 1
Generation of DENV2 fusion loop mutants via directed evolution.
(A) Alignment of top: dengue virus fusion loops; bottom: mosquito-borne flavivirus fusion loops, including yellow fever virus (YFV), Zika virus (ZIKV), West Nile virus (WNV), Kunjin virus (KUNV), …
Figure 2
Biological and physical properties of mature DENV2 fusion loop mutants.
(A) Multistep growth curves [Multiplicity of infection (MOI) = 0.05–0.1] of DV2-WT, D2-FL, and D2-FLM on C6/36 cells (left) or Vero 81 cells (right). (B) Thermostability assay on DV2-WT, D2-FL, and …
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Figure 2—source data 1
Raw gel images of the western blot shown in Figure 2C.
- https://cdn.elifesciences.org/articles/87555/elife-87555-fig2-data1-v1.zip
Figure 3
Fusion loop (FL) mutant is insensitive to FL monoclonal antibodies (mAbs), the major target for cross-reactive Abs in non-human primates (NHPs).
(A) Left: FRNT50 values for neutralization of DV2-WT, D2-FL, and D2-FLM with mAbs against the FL (1M7, 1N5, 1L6, 4G2). All Abs were tested in at least n = 3 independent experiments, except 1N5 due …
Tables
Table 1
Summary of FRNT50 values of human convalescent serum and non-human primates (NHP) infection serum against DV2, D2-FL (not determined in human serum) and D2-FLM.
FRNT50 values were derived from an eight-point dilution curve of serum started from 1:40 with subsequent threefold dilutions. FRNT50 values were calculated/extrapolated from the neutralization …
| Human # | Infected serotype | Collection tme | DV2 | D2-FL | D2-FLM |
|---|---|---|---|---|---|
| DS1500 | DENV1 | Convalescent serum | 1:75 | ND | 1:53 |
| DS2499 | 1:80 | ND | 1:52 | ||
| DS1136 | DENV3 | 1:133 | ND | 1:45 | |
| DS1160 | 1:86 | ND | 1:208 | ||
| DS0275 | DENV4 | 1:94 | ND | 1:94 | |
| DS2239 | 1:156 | ND | 1:42 | ||
| NHP # | Infected serotype | Collection time (dpi) | DV2 | D2-FL | D2-FLM |
| R628 | DENV4 | 30 | <1:40 | <1:40 | <1:40 |
| 60 | <1:40 | <1:40 | <1:40 | ||
| 90 | 1:42* | <1:40 | <1:40 | ||
| R737 | 30 | <1:40 | <1:40 | <1:40 | |
| 60 | <1:40 | <1:40 | <1:40 | ||
| 90 | <1:40 | <1:40 | <1:40 | ||
| R1132 | 30 | <1:40 | <1:40 | <1:40 | |
| 60 | <1:40 | <1:40 | <1:40 | ||
| 90 | <1:40 | <1:40 | <1:40 | ||
| R1160 | 30 | 1:58* | <1:40 | <1:40 | |
| 60 | 1:57* | 1 : 50* | <1:40 | ||
| 90 | <1:40 | 1 : 57* | 1:91* |
