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
. 2023 Nov 20;17(11):e0011770.
doi: 10.1371/journal.pntd.0011770. eCollection 2023 Nov.

EDIII-Fc induces protective immune responses against the Zika virus in mice and rhesus macaque

Hailong Su  1 , Jun Liu  1   2 , Jianhai Yu  3 , Zhenzhen Qiu  1   4 , Wenhan Liang  1   5 , Wangsheng Wu  1 , Haifeng Mo  1 , Hongwei Li  1 , Wei Zhao  3 , Weiwang Gu  6   7
Affiliations

EDIII-Fc induces protective immune responses against the Zika virus in mice and rhesus macaque

Hailong Su et al. PLoS Negl Trop Dis. .

Abstract

Zika virus can infect the fetus through the placental barrier, causing ZIKV congenital syndrome and even miscarriage, which can cause great harm to pregnant women and infants. Currently, there is no vaccine and drug available to combat the Zika virus. In this study, we designed a fusion protein named EDIII-Fc, including the EDIII region of Zika E protein and human IgG Fc fragment, and obtained 293T cells that stably secreted EDIII-Fc protein using the lentiviral expression system. Mice were immunized with the EDIII-Fc protein, and it was observed that viral replication was significantly inhibited in the immunized mice compared to non-immunized mice. In rhesus macaques, we found that EDIII-Fc effectively induce the secretion of neutralizing antibodies and T cell immunity. These experimental data provide valid data for further use of Zika virus E protein to prepare an effective, safe, affordable Zika vaccine.

PubMed Disclaimer

Conflict of interest statement

The authors have declared that no competing interest exists.

Figures

Fig 1
Fig 1. Expression of the recombinant EDIII-Fc protein.
A. Positions of EDIII protein in the E protein. The crystal structure of the ZIKV E protein dimer (PDB ID 5LBV) is shown, with EDIII of monomeric subunit A colored in red. B. Sequence Logo visualization of the EDIII alignments. Sequence Logos of the EDIII domain from multiple sequence alignments across all organisms, consisting of 1,809 protein sequences (Sequence data from Genbank, the date to obtain the sequence was to June 20, 2023). C. Schematic map of pLV-ZIKV-EDIII vector construction. D. Western blot detection EDIII expression. Line 1, cell lysate of HEK293T transfected with pLV-eGFP; Line 2, supernatant of HEK293T transfected with pLV-eGFP; Line 3, cell lysate of HEK293T transfected with pLV-ZIKV-EDIII; Line 4, supernatant of HEK293T transfected with pLV-ZIKV-EDIII.
Fig 2
Fig 2. Immunization with EDIII-Fc protects mice from ZIKV challenge.
A. Timeline for mice immunization with EDIII-Fc and blood collection. Balb/c mice were immunized with EDIII-Fc or PBS. Pre-immune blood samples were obtained 2 days before immunization, and subsequent samples were collected weekly post-immunization. B. Changes of total antibody levels in mice after immunization. Serum samples from mice immunized with EDIII-Fc (in red) or PBS (in black) were serially diluted at a 1:10 ratio. HRP-labeled sheep anti-mouse monoclonal antibody was used for ELISA detection. C. Detection of neutralizing antibody levels in mice. Two weeks after the second immunization, the sera to be tested were serially diluted 2-fold in PBS, starting from a 1:5 ratio, for plaque reduction neutralization testing. Results are reported as the fold dilution of serum that inhibited 50% of plaque formation. D. Serum viral load in mice after challenge. One week after the second immunization, seven Balb/c mice were injected subcutaneously with 104 PFU of ZIKV. Daily blood samples were collected and qPCR was carried out for viral load detection. Three replications were done in each experiment, and the experiment was repeated three times. *: P < 0.05;****: P < 0.0001. Clipart image from https://openclipart.org/.
Fig 3
Fig 3. EDIII -Fc induction the production of anti-zika antibodies in rhesus macaques A.
Timeline for rhesus macaque immunization with EDIII-Fc and blood collection. Rhesus macaques were immunized with EDIII-Fc or PBS. Pre-immune blood samples were obtained 2 days before immunization, and subsequent samples were collected weekly post-immunization. B. Changes of total antibody level in rhesus macaques after immunization. Serum samples from rhesus macaques immunized with EDIII-Fc (in red) or PBS (in black) were serially diluted at a 1:10 ratio. HRP-labeled mouse anti-monkey monoclonal antibody was used for ELISA detection. C. Detection of neutralizing antibody level in rhesus macaques. Two weeks after the third immunization, the sera to be tested were serially diluted 2-fold in PBS starting from 1:5 for plaque reduction neutralization test. Results are reported as the fold dilution of serum that inhibited 50% of plaque formation. Three replications were done in each experiment, and the experiment was repeated three times. ***: P < 0.0001. Clipart image from https://openclipart.org/.
Fig 4
Fig 4. Changes in cytokines levels secreted in rhesus macaque after immunization.
Pre-immune PBMCs and those obtained at 4 weeks, 8 weeks, and 12 weeks after immunization from rhesus macaques were incubated with ZIKV E protein for 48 hours. Following incubation, culture supernatants were collected for flow cytometric assays. Three replications were done in each experiment and repeated three times. ****: P < 0.0001.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Grossi-Soyster EN, Cook EAJ, de Glanville WA, Thomas LF, Krystosik AR, Lee J, et al.. Serological and spatial analysis of alphavirus and flavivirus prevalence and risk factors in a rural community in western Kenya. PLoS neglected tropical diseases. 2017;11(10):e0005998. Epub 2017/10/19. doi: 10.1371/journal.pntd.0005998 ; PubMed Central PMCID: PMC5659799. - DOI - PMC - PubMed
    1. Guzman H, Contreras-Gutierrez MA, Travassos da Rosa APA, Nunes MRT, Cardoso JF, Popov VL, et al.. Characterization of Three New Insect-Specific Flaviviruses: Their Relationship to the Mosquito-Borne Flavivirus Pathogens. The American journal of tropical medicine and hygiene. 2017. Epub 2017/10/11. doi: 10.4269/ajtmh.17-0350 ; PubMed Central PMCID: PMC5929187. - DOI - PMC - PubMed
    1. Figueiredo MLG, Amarilla AA, Figueiredo GG, Alfonso HL, Lippi V, Maia FGM, et al.. Cacipacore virus as an emergent mosquito-borne Flavivirus. Revista da Sociedade Brasileira de Medicina Tropical. 2017;50(4):539–42. Epub 2017/09/28. doi: 10.1590/0037-8682-0485-2016 . - DOI - PubMed
    1. Haslwanter D, Blaas D, Heinz FX, Stiasny K. A novel mechanism of antibody-mediated enhancement of flavivirus infection. PLoS pathogens. 2017;13(9):e1006643. Epub 2017/09/16. doi: 10.1371/journal.ppat.1006643 ; PubMed Central PMCID: PMC5617232. - DOI - PMC - PubMed
    1. Dick GW, Kitchen SF, Haddow AJ. Zika virus. I. Isolations and serological specificity. Transactions of the Royal Society of Tropical Medicine and Hygiene. 1952;46(5):509–20. Epub 1952/09/01. doi: 10.1016/0035-9203(52)90042-4 . - DOI - PubMed

Grants and funding

This study was financially supported by the Science and Technology Planning Project of Guangdong Province (2021B1212040016 to WG), National Key R&D Projects of China (grant 2017YFD0500600 to HL), the Science and Technology Planning Project of Guangdong Province (grant 2021B1212030014 to WZ), National Natural Science Foundation of China (82003329 and 81972924 to JL), Guangdong Basic and Applied Basic Research Foundation (2019A1515110109 to JL), China Postdoctoral Science Foundation (2019M662983 to JL). The funders had no role in the study design, data collection, analysis, decision to publish, or the preparation of the manuscript.
-