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Aedes aegypti AgBR1 antibodies modulate early Zika virus infection of mice

Abstract

A recent epidemic of Zika virus in the Americas, affecting well over a million people, caused substantial mortality and morbidity, including Guillain–Barre syndrome, microcephaly and other fetal developmental defects1,2. Preventive and therapeutic measures that specifically target the virus are not readily available. The transmission of Zika virus is predominantly mosquito-borne, and Aedes aegypti mosquitoes serve as a key vector for Zika virus3. Here, to identify salivary factors that modulate mosquito-borne Zika virus infection, we focused on antigenic proteins in mice that were repeatedly bitten by mosquitoes and developed antibodies against salivary proteins. Using a yeast surface display screen, we identified five antigenic A. aegypti salivary proteins in mice. Antiserum against one of these five proteins—A. aegypti bacteria-responsive protein 1 (AgBR1)—suppressed early inflammatory responses in the skin of mice bitten by Zika-virus-infected mosquitoes. AgBR1 antiserum also partially protected mice from lethal mosquito-borne—but not needle-injected—Zika virus infection. These data suggest that AgBR1 is a target for the prevention of mosquito-transmitted Zika virus infection.

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Fig. 1: AgBR1 identified as an antigenic protein in mice that modulates host responses in vitro and in vivo.
Fig. 2: AgBR1 antiserum protects mice from mosquito-borne Zika virus infection.
Fig. 3: AgBR1 antiserum suppresses neutrophil infiltration at the mosquito-bite site.
Fig. 4: AgBR1 antiserum modulates host responses at the mosquito-bite site.

Data availability

The data that support the findings of this study are available from the corresponding authors on request. The RNA-seq data have been deposited in the Gene Expression Omnibus under the accession number GSE125194.

References

  1. 1.

    Ventura, C. V., Maia, M., Bravo-Filho, V., Gois, A. L. & Belfort, R. Jr Zika virus in Brazil and macular atrophy in a child with microcephaly. Lancet 387, 228 (2016).

    Article  Google Scholar 

  2. 2.

    Olsen, B. & Lundkvist, A. Zika virus—ancient virus gets new life in a new ecosystem. Microcephaly and Guillain–Barre syndrome are possible consequences when there is no background herd immunity in the population (in Swedish). Lakartidningen 113, DX9X (2016).

    PubMed  Google Scholar 

  3. 3.

    Musso, D. & Gubler, D. J. Zikavirus. Clin. Microbiol. Rev. 29, 487–524 (2016).

    CAS  Article  Google Scholar 

  4. 4.

    Coutinho-Abreu, I. V., Guimaraes-Costa, A. B. & Valenzuela, J. G. Impact of insect salivary proteins in blood feeding, host immunity, disease, and in the development of biomarkers for vector exposure. Curr. Opin. Insect Sci. 10, 98–103 (2015).

    Article  Google Scholar 

  5. 5.

    Fontaine, A. et al. Implication of haematophagous arthropod salivary proteins in host-vector interactions. Parasit. Vectors 4, 187 (2011).

    CAS  Article  Google Scholar 

  6. 6.

    Hayashi, H. et al. Anopheline anti-platelet protein from a malaria vector mosquito has anti-thrombotic effects in vivo without compromising hemostasis. Thromb. Res. 129, 169–175 (2012).

    CAS  Article  Google Scholar 

  7. 7.

    Pingen, M. et al. Host inflammatory response to mosquito bites enhances the severity of arbovirus infection. Immunity 44, 1455–1469 (2016).

    CAS  Article  Google Scholar 

  8. 8.

    Ruckert, C. & Ebel, G. D. How do virus-mosquito interactions lead to viral emergence? Trends Parasitol. 34, 310–321 (2018).

    Article  Google Scholar 

  9. 9.

    Conway, M. J. et al. Mosquito saliva serine protease enhances dissemination of dengue virus into the mammalian host. J. Virol. 88, 164–175 (2014).

    Article  Google Scholar 

  10. 10.

    Conway, M. J. et al. Aedes aegypti D7 saliva protein inhibits dengue virus infection. PLoS Negl. Trop. Dis. 10, e0004941 (2016).

    Article  Google Scholar 

  11. 11.

    Jin, L. et al. Salivary factor LTRIN from Aedes aegypti facilitates the transmission of Zika virus by interfering with the lymphotoxin-β receptor. Nat. Immunol. 19, 342–353 (2018).

    CAS  Article  Google Scholar 

  12. 12.

    Wasinpiyamongkol, L. et al. Blood-feeding and immunogenic Aedes aegypti saliva proteins. Proteomics 10, 1906–1916 (2010).

    CAS  Article  Google Scholar 

  13. 13.

    Rabilloud, T., Chevallet, M., Luche, S. & Lelong, C. Two-dimensional gel electrophoresis in proteomics: past, present and future. J. Proteomics 73, 2064–2077 (2010).

    CAS  Article  Google Scholar 

  14. 14.

    Schuijt, T. J. et al. Identification and characterization of Ixodes scapularis antigens that elicit tick immunity using yeast surface display. PLoS ONE 6, e15926 (2011).

    CAS  Article  Google Scholar 

  15. 15.

    Lee, C. G. et al. Role of chitin and chitinase/chitinase-like proteins in inflammation, tissue remodeling, and injury. Annu. Rev. Physiol. 73, 479–501 (2011).

    CAS  Article  Google Scholar 

  16. 16.

    Ribeiro, J. M. et al. An annotated catalogue of salivary gland transcripts in the adult female mosquito, Aedes aegypti. BMC Genom. 8, 6 (2007).

    Article  Google Scholar 

  17. 17.

    Beatty, P. R. et al. Dengue virus NS1 triggers endothelial permeability and vascular leak that is prevented by NS1 vaccination. Sci. Transl. Med. 7, 304ra141 (2015).

    Article  Google Scholar 

  18. 18.

    Tanaka, T. & Kishimoto, T. The biology and medical implications of interleukin-6. Cancer Immunol. Res. 2, 288–294 (2014).

    CAS  Article  Google Scholar 

  19. 19.

    Calvo, E., Mans, B. J., Andersen, J. F. & Ribeiro, J. M. Function and evolution of a mosquito salivary protein family. J. Biol. Chem. 281, 1935–1942 (2006).

    CAS  Article  Google Scholar 

  20. 20.

    Rathore, A. P. S. & St John, A. L. Immune responses to dengue virus in the skin. Open Biol. 8, 18087 (2018).

  21. 21.

    Kieny, M. P., Excler, J. L. & Girard, M. Research and development of new vaccines against infectious diseases. Am. J. Public Health 94, 1931–1935 (2004).

    Article  Google Scholar 

  22. 22.

    Sarathy, V. V., Milligan, G. N., Bourne, N. & Barrett, A. D. Mouse models of dengue virus infection for vaccine testing. Vaccine 33, 7051–7060 (2015).

    CAS  Article  Google Scholar 

  23. 23.

    Zhu, J., Huang, X. & Yang, Y. Type I IFN signaling on both B and CD4 T cells is required for protective antibody response to adenovirus. J. Immunol. 178, 3505–3510 (2007).

    CAS  Article  Google Scholar 

  24. 24.

    Fong, S. W., Kini, R. M. & Ng, L. F. P. Mosquito saliva reshapes alphavirus infection and immunopathogenesis. J. Virol. 92, e01004-17 (2018).

  25. 25.

    Shi, C. & Pamer, E. G. Monocyte recruitment during infection and inflammation. Nat. Rev. Immunol. 11, 762–774 (2011).

    CAS  Article  Google Scholar 

  26. 26.

    Osuna, C. E. et al. Zika viral dynamics and shedding in rhesus and cynomolgus macaques. Nat. Med. 22, 1448–1455 (2016).

    CAS  Article  Google Scholar 

  27. 27.

    Bai, F. et al. A paradoxical role for neutrophils in the pathogenesis of West Nile virus. J. Infect. Dis. 202, 1804–1812 (2010).

    CAS  Article  Google Scholar 

  28. 28.

    Dudley, D. M. et al. Infection via mosquito bite alters Zika virus tissue tropism and replication kinetics in rhesus macaques. Nat. Commun. 8, 2096 (2017).

    Article  Google Scholar 

  29. 29.

    Carman, W. F. et al. Vaccine-induced escape mutant of hepatitis B virus. Lancet 336, 325–329 (1990).

    CAS  Article  Google Scholar 

  30. 30.

    Uraki, R., Hastings, A. K., Gloria-Soria, A., Powell, J. R. & Fikrig, E. Altered vector competence in an experimental mosquito-mouse transmission model of Zika infection. PLoS Negl. Trop. Dis. 12, e0006350 (2018).

    Article  Google Scholar 

  31. 31.

    Chao, G. et al. Isolating and engineering human antibodies using yeast surface display. Nat. Protoc. 1, 755–768 (2006).

    CAS  Article  Google Scholar 

  32. 32.

    Uraki, R. et al. Zika virus causes testicular atrophy. Sci. Adv. 3, e1602899 (2017).

    Article  Google Scholar 

  33. 33.

    Balaban, A. E., Neuman, K., Sinnis, P. & Balaban, R. S. Robust fluorescent labelling of micropipettes for use in fluorescence microscopy: application to the observation of a mosquito borne parasite infection. J. Microsc. 269, 78–84 (2018).

    CAS  Article  Google Scholar 

  34. 34.

    Subramanian, A. et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl Acad. Sci. USA 102, 15545–15550 (2005).

    CAS  Article  Google Scholar 

  35. 35.

    Nakajima, S. et al. Prostaglandin I2-IP signaling promotes Th1 differentiation in a mouse model of contact hypersensitivity. J. Immunol. 184, 5595–5603 (2010).

    CAS  Article  Google Scholar 

  36. 36.

    Chang, Q., Ornatsky, O. & Hedley, D. Staining of frozen and formalin-fixed, paraffin-embedded tissues with metal-labeled antibodies for imaging mass cytometry analysis. Curr. Protoc. Cytom. 82, 12.47.1–12.47.8 (2017).

    Article  Google Scholar 

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Acknowledgements

We thank S. Shresta at the La Jolla Institute for Allergy and Immunology for originally providing us with the AG129 mouse strain. We also thank S. Ren, Y.-M. Chuang, S. Householder, S. Stanley, H. Sproch and B. V. Wyk for supporting experiments and analysing data. The imaging mass cytometry was conducted at the Yale CyTOF facility and the RNA-seq service was conducted at the Yale Stem Cell Center Genomics Core facility, which was supported by the Connecticut Regenerative Medicine Research Fund and the Li Ka Shing Foundation. This work was supported by NIH grant nos AI089992 and AI127865, and the Japan Society for the Promotion of Science Overseas Research Fellowships. E.F. and A.I. are investigators with the Howard Hughes Medical Institute.

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Contributions

R.U. and E.F. designed the experiments. R.U. performed the majority of the experiments and analysed the data. A.K.H., A.M.-L. and J.R.G. assisted in the experiments with mosquitoes. R.U. and T.S. performed the analysis of the RNA-seq. T.T. performed the histopathological analyses. A.I., D.A.H. and R.R.M. contributed experimental suggestions and strengthened the writing of the manuscript. R.U. and E.F wrote the manuscript. All authors reviewed, critiqued and provided comments on the text.

Corresponding author

Correspondence to Erol Fikrig.

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The authors declare no competing interests.

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Supplementary information

Supplementary Information

Supplementary Figures 1–12, Supplementary Tables 1–3 and Supplementary Table 6.

Reporting Summary

Supplementary Table 4

List of upregulated genes at the bite site of mice treated with control (naïve) serum.

Supplementary Table 5

List of GSEA-enriched pathway at bite sites in mice treated with control serum using hallmark gene sets (http://software.broadinstitute.org/gsea/msigdb/genesets.jsp?collection=H).

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Uraki, R., Hastings, A.K., Marin-Lopez, A. et al. Aedes aegypti AgBR1 antibodies modulate early Zika virus infection of mice. Nat Microbiol 4, 948–955 (2019). https://doi.org/10.1038/s41564-019-0385-x

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