Antigenic mapping and functional characterization of human New World hantavirus neutralizing antibodies
- Department of Pathology, Microbiology and Immunology, Vanderbilt University, United States
- Vanderbilt Vaccine Center, Vanderbilt University Medical Center, United States
- Virology Division, United States Army Medical Research Institute of Infectious Diseases, United States
- Department of Pathology, The University of Texas Medical Branch at Galveston, United States
- Galveston National Laboratory, United States
- Institut Pasteur, Université Paris Cité, France
- Department of Biochemistry and Mass Spectrometry Research Center, Vanderbilt University, United States
- Department of Chemistry, Vanderbilt University, United States
- Department of Microbiology and Immunology, University of Texas Medical Branch, United States
- Department of Pediatrics, Vanderbilt University Medical Center, United States
Figures
Figure 1
Hantavirus neutralizing antibodies target four distinct regions on the glycoprotein spike.
(a) Binding potency of mAbs to recombinant hantavirus antigens, ANDV GnH/Gc, ANDV GnH/Gc_H953F, MAPV GnH/Gc, ANDV GnH, ANDV GnB, ANDV Gc, expressed in S2 cells. Binding curves were obtained using …
Figure 2 with 2 supplements
Escape mutant generation and mutagenesis mapping indicate critical binding residues for hantavirus mAbs.
(a) Results from viral escape selection for indicated antibodies. Real-time cellular analysis escape mutant mapping shows the number of replicates with escape over the total number of replicates for …
-
Figure 2—source data 1
Percent mAb binding in the presence of ANDV mutants.
- https://cdn.elifesciences.org/articles/81743/elife-81743-fig2-data1-v2.docx
-
Figure 2—source data 2
Percent mAb binding the presence of SNV mutants.
- https://cdn.elifesciences.org/articles/81743/elife-81743-fig2-data2-v2.docx
Figure 2—figure supplement 1
Mutagenesis expression levels and gating strategy.
(a) Neutralization of escape mutant viruses to antibodies at a saturating concentration (10 µg/mL). The data shown are averages ± SD from three experiments, n=9. (b) Expression levels of Gn/Gc point …
Figure 2—figure supplement 2
Amino acid alignment of hantavirus species.
Multiple sequence alignment of the M-segment from six representative Orthohantaviruses. Domains are colored as followed, Gn domain A: purple, domain B: dark purple, β-ribbon domain: light purple, …
Figure 3 with 2 supplements
BnAbs SNV-53 and SNV-24 target two sites on the GnH/Gc heterodimer.
(a) Representative nsEM 2D-class averages of SNV-53 and SNV-24 Fabs in complex with MAPV GnH/Gc heterodimer. (b) Surface representations (light grey) of SNV-53 (blue) and SNV-24 (green) in complex …
Figure 3—figure supplement 1
SNV-53 Fab docked to the hantavirus surface glycoprotein lattice (EMD-11236).
Model is colored as follows, Gn: red, Gc: yellow, SNV-53 Fab: green/blue.
Figure 3—figure supplement 2
Hydrogen-deuterium exchange mass spectrometry analysis of hantavirus antibodies in complex with ANDV GnH/Gc.
Relative fractional deuterium uptake difference at the 5000 s time point was mapped onto the cryo-EM reconstruction of the glycoprotein complex (PDB: 6ZJM). The relative fractional uptake difference …
Figure 4 with 2 supplements
Cryo-EM structure of neutralizing antibodies ANDV-5 and ANDV-34 in complex with ANDV GnH.
(a) Top view (right) and side view (left) of the heterotetramer Gn/Gc (Serris et al., 2020) (PDB: 6ZJM) with low resolution map and model of Gn(H) ANDV-5 and ANDV-34 (purple, orange and red, …
Figure 4—figure supplement 1
Workflow of cryo-electron microscopy processing for model of ANDV-5 and ANDV-34 Fabs in complex with ANDV GnH.
Figure 4—figure supplement 2
Residue interaction plot of GnH with ANDV-5 or ANDV-34.
Figure 5
Potently neutralizing hantavirus mAbs inhibit viral entry through viral attachment blocking and/or fusion inhibition.
(a) Neutralization curves of IgG1 and Fab forms of broad mAbs (SNV-53, ANDV-44 or SNV-24) to VSV/SNV and VSV/ANDV determined through real-time cellular analysis. The data shown are representative …
Figure 6
Reactivity and potency of germline revertant forms of NWH antibodies.
(a) Representative binding curves for all germline reverted forms of SNV-53 and ANDV-44 bnAbs to Expi293F cells transfected with ANDV, SNV, PUUV, DOBV, HTNV, or SEOV Gn/Gc. The value for % PE+ cells …
Figure 7 with 1 supplement
SNV-53 protects hamsters when given before or after HTNV inoculation.
(a) 8-week-old Syrian hamsters (n=8 per treatment group) were administered 5 mg/kg of the indicated Ab treatment and then inoculated 1 day later with 200 PFU of HTNV i.m. All animals were sacrificed …
Figure 7—figure supplement 1
Pre- and post-exposure HTNV serum PsVNA50 data and pathology staining.
(a) Neutralizing antibody levels in serum for the four treatment groups at days 0 or 28 measured by PsVNA50 from the pre-exposure HTNV study. (b) In situ hybridization (ISH) staining scores for …
Figure 8
ANDV-34 and ANDV-5 protect Syrian golden hamsters for lethal ANDV challenge.
(a) Eight-week-old Syrian hamsters (n=6 per treatment group) were inoculated with 200 PFU of ANDV i.m., and 10 mg/kg of indicated mAb was administered via i.p. route at 2 and 5 dpi. Animals were …
Author response image 1
Author response image 2
Author response image 3
Author response image 4
Author response image 5
Author response image 6
Author response image 7
Author response image 8
Author response image 9
Assay values for individual animals.
Individual animal PsVNA50 (log 10) titers were sorted highest to lowest and plotted. Day 28 ELISA titers (log 10), PCR levels (Log 10), and ISH scores (0-3) were also plotted. (A) Data from …
Author response image 10
Relationship between level of neutralizing antibodies in serum and protection for pre-exposure experiment.
There is a negative correlation for neutralizing antibody levels and infection for all endpoints measured. When antibody is administered 1 day pre-exposure, the level of neutralizing antibody …
Author response image 11
Relationship between level of neutralizing antibodies in serum and protection for post-exposure experiment.
There is a negative correlation for neutralizing antibody levels and infection for all endpoints measured. When antibody is administered 3 days post-exposure, the level of neutralizing antibody …
Additional files
-
Supplementary file 1
Summary table of electron microscopy statistics.
- https://cdn.elifesciences.org/articles/81743/elife-81743-supp1-v2.docx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/81743/elife-81743-mdarchecklist1-v2.docx
