Ephrin receptor A2 is a functional entry receptor for Epstein–Barr virus


Epstein–Barr virus (EBV) is an oncogenic virus that infects more than 90% of the world’s population1. EBV predominantly infects human B cells and epithelial cells, which is initiated by fusion of the viral envelope with a host cellular membrane2. The mechanism of EBV entry into B cells has been well characterized3. However, the mechanism for epithelial cell entry remains elusive. Here, we show that the integrins αvβ5, αvβ6 and αvβ8 do not function as entry and fusion receptors for epithelial cells, whereas Ephrin receptor tyrosine kinase A2 (EphA2) functions well for both. EphA2 overexpression significantly increased EBV infection of HEK293 cells. Using a virus-free cell–cell fusion assay, we found that EphA2 dramatically promoted EBV but not herpes simplex virus (HSV) fusion with HEK293 cells. EphA2 silencing using small hairpin RNA (shRNA) or knockout by CRISPR–Cas9 blocked fusion with epithelial cells. This inhibitory effect was rescued by the expression of EphA2. Antibody against EphA2 blocked epithelial cell infection. Using label-free surface plasmon resonance binding studies, we confirmed that EphA2 but not EphA4 specifically bound to EBV gHgL and this interaction is through the EphA2 extracellular domain (ECD). The discovery of EphA2 as an EBV epithelial cell receptor has important implications for EBV pathogenesis and may uncover new potential targets that can be used for the development of novel intervention strategies.

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Fig. 1: Identification of EphA2 as the potential EBV epithelial cell receptor.
Fig. 2: EphA2 can promote both EBV infection and virus-free cell–cell fusion.
Fig. 3: EphA2 is essential for EBV infection and virus-free cell–cell fusion.
Fig. 4: EphA2 binds to EBV gHgL with low affinity.


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The authors acknowledge help and advice from members of the Longnecker and Jardetzky laboratories, especially N. Susmarski. The authors thank M. Lingen and T. Li from the Human Tissue Resource Center (HTRC) at the University of Chicago for performing experiments during the revision, and M. Manzano for providing reagents. The research was supported by AI076183 (to R.L. and T.J.) from the National Institute of Allergy and Infectious Diseases, by CA117794 (to R.L. and T.J.) from the National Cancer Institute, by AI119480 (to R.L., H.Z. and T.J.) from the National Institute of Allergy and Infectious Diseases as well as by the Chicago Biomedical Consortium (Fall 14-0121 and 15-0257, to J.C.).

Author information

J.C. and R.L. designed the overall study, with input from the co-authors. J.C. performed the key experiments. K.S. performed the initial microarray analysis, cloning gH and gL expression plasmids, purified protein preparations and carried out EphA2 binding experiments. X.Z. performed the RNA-seq analysis and helped with the sgRNA constructs and statistical analysis. S.S. helped with cell cultures. B.E.P.W. contributed key reagents. J.C. and R.L. wrote the manuscript. X.Z., K.S. and T.S.J. contributed expertise and helped write the paper. All authors analysed the results, read and approved the manuscript for submission.

Correspondence to Richard Longnecker.

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Supplementary information

Supplementary Information

Supplementary Figures 1–6, Supplementary Table 5.

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Supplementary Table 1

Candidate receptor genes identified by comparing GeneRNA seq data from HEK293 and B cells (HEK293>5 FPKM, B< 5 FPKM, HEK293/B >10, membrane protein).

Supplementary Table 2

Candidate receptor genes identified by comparing GeneRNA seq data from HEK293 and B cells (HEK293>5 FPKM, B< 5 FPKM, HEK293/B >10, membrane protein, AGS/HEK293>2).

Supplementary Table 3

Candidate receptor genes identified by comparing GeneRNA seq data from AGS and B cells (AGS>5 FPKM, B< 5 FPKM, AGS/B >10, membrane protein).

Supplementary Table 4

Candidate receptor genes identified by comparing RNA seq from AGS and B cells (AGS>5 FPKM, B< 5 FPKM, AGS/B >10, membrane protein, AGS/HEK293>2).

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