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Ephrin receptor A2 is an epithelial cell receptor for Epstein–Barr virus entry

Nature Microbiology volume 3, pages 1–8 (2018)Cite this article

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An Author Correction to this article was published on 20 April 2018

Abstract

Epstein–Barr virus (EBV) is causally associated with nasopharyngeal carcinoma, 10% of gastric carcinoma and various B cell lymphomas1. EBV infects both B cells and epithelial cells2. Recently, we reported that epidermal growth factor and Neuropilin 1 markedly enhanced EBV entry into nasopharyngeal epithelial cells3. However, knowledge of how EBV infects epithelial cells remains incomplete. To understand the mechanisms through which EBV infects epithelial cells, we integrated microarray and RNA interference screen analyses and found that Ephrin receptor A2 (EphA2) is important for EBV entry into the epithelial cells. EphA2 short interfering RNA knockdown or CRISPR–Cas9 knockout markedly reduced EBV epithelial cell infection, which was mostly restored by EphA2 complementary DNA rescue. EphA2 overexpression increased epithelial cell EBV infection. Soluble EphA2 protein, antibodies against EphA2, soluble EphA2 ligand EphrinA1, or the EphA2 inhibitor 2,5-dimethylpyrrolyl benzoic acid derivative efficiently blocked EBV epithelial cell infection. Mechanistically, EphA2 interacted with EBV entry proteins gH/gL and gB to facilitate EBV internalization and fusion. The EphA2 Ephrin-binding domain and fibronectin type III repeats domain were essential for EphA2-mediated EBV infection, while the intracellular domain was dispensable. This is distinct from Kaposi’s sarcoma-associated herpesvirus infection through EphA24. Taken together, our results identify EphA2 as a critical player for EBV epithelial cell entry.

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Fig. 1: EphA2 promotes EBV infection of epithelial cells.
Fig. 2: EphA2 interacts with EBV glycoprotein gH/gL and gB.
Fig. 3: Antagonizing EphA2 ectodomain function markedly impairs EBV infection of epithelial cells.
Fig. 4: EphA2 facilitates EBV internalization and fusion.

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Acknowledgements

This study was supported by the National Key R&D Program (2016YFA0502100 and 2017YFA0505600), the National Natural Science Foundation of China (81520108022, 81502374, 81230045, 81372244, 81572600), the Science and Technology project of Guangdong Province (2014B050504004, 2015B050501005), the talent program of Guangdong Province (412022693047) and the Science and Technology project of Guangdong Province (2014B050504004, 2015B050501005). PlasmidspCAGT7 and pT7EMCLuc were kindly provided by Professor Richard Longnecker and Patricia G. Spear (Northwestern University). Plasmid p2670 was generously gifted from Professor Wolfgang Hammerschmidt (Helmholtz Zentrum München). B.E.G. is supported by a Burroughs Wellcome Foundation Career Award in Medical Sciences. The authenticity of this article has been validated by uploading the key raw data onto the Research Data Deposit public platform (www.researchdata.org.cn), with the approval number RDDB2017000231.

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  1. Hua Zhang, Yan Li and Hong-Bo Wang contributed equally to this study

Authors and Affiliations

  1. Sun Yat-sen University Cancer Center; State Key Laboratory of Oncology, South China; Collaborative Innovation Center for Cancer Medicine, Guangzhou, China

    Hua Zhang, Ao Zhang, Mei-Ling Chen, Zhi-Xin Fang, Xiao-Dong Dong, Shi-Bing Li, Yong Du, Dan Xiong, Jiang-Yi He, Man-Zhi Li, Yan-Min Liu, Ai-Jun Zhou, Qian Zhong, Yi-Xin Zeng & Mu-Sheng Zeng

  2. Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, China

    Yan Li

  3. Guangdong Provincial Key Laboratory of Liver Disease Research, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China

    Hong-Bo Wang

  4. Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Elliott Kieff, Benjamin E. Gewurz & Bo Zhao

  5. Immunobiology and Transplant Science Center, Houston Methodist Research Institute, Houston, TX, USA

    Zhiqiang Zhang

  6. Department of Surgery, Weill Cornell Medical College of Cornell University, New York, NY, USA

    Zhiqiang Zhang

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Contributions

M.-S.Z., B.Z., and Z.Q.Z. conceived and designed the experiments, provided supervision and wrote the manuscript. H.Z. conceived the experiments. H.Z., Y.L. and H.-B.W. performed, analysed the key experiments and wrote the manuscript. A.Z., M.-L.C., Z.-X.F., J-Y.H. and X.D.D. performed the experiments. Y.Z.Z. Q.Z., S-B.L, D.X. and Y.D. designed and analysed the data. M.-Z.L., A.-J.Z., E.K., B.G. and Y.-M.L. contributed reagents and materials. All authors read and approved the final version of the manuscript.

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Correspondence to Bo Zhao or Mu-Sheng Zeng.

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Zhang, H., Li, Y., Wang, HB. et al. Ephrin receptor A2 is an epithelial cell receptor for Epstein–Barr virus entry. Nat Microbiol 3, 1–8 (2018). https://doi.org/10.1038/s41564-017-0080-8

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