Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Dec 8;185(25):4826-4840.e17.
doi: 10.1016/j.cell.2022.10.023. Epub 2022 Nov 18.

A Zika virus-specific IgM elicited in pregnancy exhibits ultrapotent neutralization

Tulika Singh  1 , Kwan-Ki Hwang  2 , Andrew S Miller  3 , Rebecca L Jones  2 , Cesar A Lopez  4 , Sarah J Dulson  4 , Camila Giuberti  5 , Morgan A Gladden  2 , Itzayana Miller  6 , Helen S Webster  2 , Joshua A Eudailey  6 , Kan Luo  2 , Tarra Von Holle  2 , Robert J Edwards  2 , Sarah Valencia  2 , Katherine E Burgomaster  7 , Summer Zhang  8 , Jesse F Mangold  2 , Joshua J Tu  2 , Maria Dennis  2 , S Munir Alam  2 , Lakshmanane Premkumar  4 , Reynaldo Dietze  9 , Theodore C Pierson  7 , Eng Eong Ooi  8 , Helen M Lazear  4 , Richard J Kuhn  3 , Sallie R Permar  10 , Mattia Bonsignori  11
Affiliations

A Zika virus-specific IgM elicited in pregnancy exhibits ultrapotent neutralization

Tulika Singh et al. Cell. .

Abstract

Congenital Zika virus (ZIKV) infection results in neurodevelopmental deficits in up to 14% of infants born to ZIKV-infected mothers. Neutralizing antibodies are a critical component of protective immunity. Here, we demonstrate that plasma IgM contributes to ZIKV immunity in pregnancy, mediating neutralization up to 3 months post-symptoms. From a ZIKV-infected pregnant woman, we isolated a pentameric ZIKV-specific IgM (DH1017.IgM) that exhibited ultrapotent ZIKV neutralization dependent on the IgM isotype. DH1017.IgM targets an envelope dimer epitope within domain II. The epitope arrangement on the virion is compatible with concurrent engagement of all ten antigen-binding sites of DH1017.IgM, a solution not available to IgG. DH1017.IgM protected mice against viremia upon lethal ZIKV challenge more efficiently than when expressed as an IgG. Our findings identify a role for antibodies of the IgM isotype in protection against ZIKV and posit DH1017.IgM as a safe and effective candidate immunotherapeutic, particularly during pregnancy.

Keywords: IgM; Zika; cross-linking; immunotherapy; isotype; multivalent binding; neutralization; pregnancy; therapeutic antibodies.

PubMed Disclaimer

Conflict of interest statement

Declaration of interests M.B., S.R.P., T.S., and K.-K.H. have filed a patent application directed to antibodies that are related to this work. S.R.P. serves as a consultant to Moderna, Merck, Pfizer, GSK, Hoopika, and Dynavax CMV vaccine programs and leads a sponsored research program with Moderna and Merck.

Figures

Figure 1.
Figure 1.. Contribution of plasma IgM to ZIKV neutralization from acute infection through late convalescence.
A. Plasma IgM ZIKV binding across ZIKV-infected mothers (n=10) from 8 to 406 days post symptoms (DPS). Plasma was tested at 1:10 dilution in two replicates. Error bars: standard deviation (SD). Dotted line: limit of detection. B. Percent ZIKV neutralization attributable to plasma IgM. Each sample was tested in three replicates. C. Comparison of early (<100 DPS) and late (≥100 DPS) plasma IgM-mediated ZIKV neutralization. When multiple samples from the same individual were collected within each time frame, only peak responses are shown. Statistical significance was measured using an unpaired Mann-Whitney test. P value is shown. See also Table S1 and Figure S1.
Figure 2:
Figure 2:. Frequencies of ZIKV-reactive B cells.
A. Frequency of peripheral blood ZIKV-binding unfractionated and memory B cells. Days-post symptoms (DPS) of PBMC collection are indicated for each pregnant woman. B. Proportion of ZIKV-reactive memory B cells (CD3−/CD14−/CD16−/CD19+/IgD−/ZIKV+) over total B cells (CD3−/CD14−/CD16−/CD19+/ZIKV+). C. Immunoglobulin (Ig) concentration in supernatants of cultured unfractionated (left) and memory (right) B cells, clustered by isotype. Red circles: cultures for which ZIKV specificity was confirmed (OD450 range = 0.44-2.5).
Figure 3.
Figure 3.. Characterization of B-LCL-derived ZIKV-specific monoclonal antibodies.
A. Left: DH1017.IgM native PAGE gel in non-reducing conditions. Ladder on the left. Right: Negative stain electron microscopy of purified DH1017.IgM showing representative class averages of hexameric and pentameric particles. Scale bar is 40nm. B. Binding to whole ZIKV PRVABC59 virions by B-LCL-derived mAbs (n=9). DH1017.IgM is shown in red. Error bars: SD of two replicates. Data representative of duplicate experiments. C. Heatmap showing binding to ZIKV and DENV serotypes 1-4 virions by the nine B-LCL-derived mAbs. Palivizumab: negative control. D. ZIKV PRVABC59 neutralization by the 9 B-LCL-derived mAbs expressed as percentage of the number of foci relative to the virus alone condition. Dotted line: 50% viral inhibition. Error bars: SD of three replicates. Data representative of duplicate experiments. E. ZIKV PRVABC59 strain neutralization curves of DH1017.IgM (red) and 9 previously described IgG ZIKV neutralizing mAbs. Error bars: SD of three replicates. Data representative of duplicate experiments. See also Figure S2.
Figure 4.
Figure 4.. Functional characterization of DH1017.IgM, DH1017.IgG, and DH1017.Fab.
ZIKV virion binding of DH1017.Fab (A), DH1017.IgG (B) and DH1017.IgM (C). Error bars: SD of two experiments with 2 (DH1017.Fab) or 6 (DH1017.IgM and DH1017.IgG) replicates each. D. ZIKV neutralization of Fab, IgG and IgM versions of DH1017. Dotted line: 50% relative infection compared to virus alone (FRNT50). Error bars: SD from 3 replicates. Data representative of duplicate experiments. E. DH1017.IgM and DH1017.IgG-mediated FRNT50 (y-axis) over increasing complement concentrations supplemented through Normal Human Serum (NHS) (x-axis). MAbs were run in triplicate. F, G. Antibody-dependent enhancement (ADE) of infection using ZIKV H/PF/2013 reporter virus particles on K562 (F) and THP-1 (G) cells. Data representative of at least three experiments. H. ADE tested on THP1.2S cells measured by plaque assay. Error bars: SD from 6 replicates. Dotted line: virus-only control (grey area: ≤1 SD). See also Figure S3.
Figure 5:
Figure 5:. DH1017.IgM protects mice more efficiently than DH1017.IgG against viremia upon lethal ZIKV challenge.
A. Serum viral load in ZIKV infected 5-week-old Ifnar1−/− mice treated with DH1017.IgM at 103 pmol/dose (n=7) or 52 pmol/dose (n=5), or non-ZIKV reactive mAb DH1036.IgM (103 pmol/dose, n=4). Line at mean. Dotted line at limit of detection. B. Survival curves for each IgM intervention group. C. Human IgM serum concentrations in mice treated with 103 pmol/dose of DH1017.IgM or DH1036.IgM measured up to 10 days post-infection. D. Serum viral load in mice receiving of DH1017.IgG at 103 pmol/dose (n=8) or 515 pmol/dose (n=4). Additional 4 mice treated with 103 pmol/dose DH1017.IgM are shown. Line at mean. Dotted line at limit of detection.
Figure 6.
Figure 6.. DH1017 clone interacts with envelope dimer.
A. Surface-shaded view of the Zika virion bound with the DH1017 Fab. Shading represents the distance to the virion center (scale bar in Å). Black triangle: asymmetric unit; pentagon: five-fold axis; triangle: three-fold axis; oval: two-fold axis. B. Surface representation of the ZIKV E ectodomain asymmetric unit (PDB 6CO8) shown in top view unbound (left), bound with the DH1017.Fab variable (cyan) and constant (green) domains (middle), and in bound side view (right). E ectodomain DI: red, DII: yellow, and DIII: blue. DH1017.Fab footprint residues: magenta. C. Radially colored roadmap (scale bar in Å). The icosahedral and quasi two-fold axes are labelled i2f and q2f, respectively. The monomer chains of two E dimers are labeled A’ and A at the i2f, and C and E at the q2f. Residues on the surface of the virus within 6 Å of the variable domain structure fit to the density map are colored yellow on chain C at the q2f axis and magenta on chain A’ and A at the i2f axis. D. Fab DH1017 epitope shown on the primary sequence of E ectodomain. Epitope residues: magenta. Domains DI, DII, and DIII: red, yellow and blue lines, respectively. See also Figures S4, S5, S6 and Table S3.
See this image and copyright information in PMC

Comment in

  • Neutralizing Zika virus.
    Crunkhorn S. Crunkhorn S. Nat Rev Drug Discov. 2023 Jan;22(1):19. doi: 10.1038/d41573-022-00201-3. Nat Rev Drug Discov. 2023. PMID: 36450861 No abstract available.

References

    1. Abbink P, Larocca RA, De La Barrera RA, Bricault CA, Moseley ET, Boyd M, Kirilova M, Li Z, Ng’ang’a D, Nanayakkara O, et al. (2016). Protective efficacy of multiple vaccine platforms against Zika virus challenge in rhesus monkeys. Science. 353, 1129–1132. 10.1126/science.aah6157. - DOI - PMC - PubMed
    1. Baronti C, Piorkowski G, Charrel RN, Boubis L, Leparc-Goffart I, and de Lamballerie X (2014). Complete coding sequence of Zika virus from a French Polynesia outbreak in 2013. Genome Announc. 2, 500–514. 10.1128/genomeA.00500-14. - DOI - PMC - PubMed
    1. Bonsignori M, Hwang K-K, Chen X, Tsao C-Y, Morris L, Gray E, Marshall DJ, Crump JA, Kapiga SH, Sam NE, et al. (2011). Analysis of a Clonal Lineage of HIV-1 Envelope V2/V3 Conformational Epitope-Specific Broadly Neutralizing Antibodies and Their Inferred Unmutated Common Ancestors. J. Virol 85, 9998–10009. 10.1128/JVI.05045-11. - DOI - PMC - PubMed
    1. Bonsignori M, Wiehe K, Grimm SK, Lynch R, Yang G, Kozink DM, Perrin F, Cooper AJ, Hwang KK, Chen X, et al. (2014). An autoreactive antibody from an SLE/HIV-1 individual broadly neutralizes HIV-1. J. Clin. Invest 124, 1835–1843. 10.1172/JCI73441. - DOI - PMC - PubMed
    1. Bonsignori M, Zhou T, Sheng Z, Chen L, Gao F, Joyce MG, Ozorowski G, Chuang GY, Schramm CA, Wiehe K, et al. (2016). Maturation Pathway from Germline to Broad HIV-1 Neutralizer of a CD4-Mimic Antibody. Cell 165, 449–463. 10.1016/j.cell.2016.02.022. - DOI - PMC - PubMed