Abstract
Kaposi's sarcoma–associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma1, a highly vascularized tumor originating from lymphatic endothelial cells, and of at least two different B cell malignancies2,3. A dimeric complex formed by the envelope glycoproteins H and L (gH-gL) is required for entry of herpesviruses into host cells4. We show that the ephrin receptor tyrosine kinase A2 (EphA2) is a cellular receptor for KSHV gH-gL. EphA2 co-precipitated with both gH-gL and KSHV virions. Infection of human epithelial cells with a GFP-expressing recombinant KSHV strain, as measured by FACS analysis, was increased upon overexpression of EphA2. Antibodies against EphA2 and siRNAs directed against EphA2 inhibited infection of endothelial cells. Pretreatment of KSHV with soluble EphA2 resulted in inhibition of KSHV infection by up to 90%. This marked reduction of KSHV infection was seen with all the different epithelial and endothelial cells used in this study. Similarly, pretreating epithelial or endothelial cells with the soluble EphA2 ligand ephrinA4 impaired KSHV infection. Deletion of the gene encoding EphA2 essentially abolished KSHV infection of mouse endothelial cells. Binding of gH-gL to EphA2 triggered EphA2 phosphorylation and endocytosis, a major pathway of KSHV entry5,6. Quantitative RT-PCR and in situ histochemistry revealed a close correlation between KSHV infection and EphA2 expression both in cultured cells derived from human Kaposi's sarcoma lesions or unaffected human lymphatic endothelium, and in situ in Kaposi's sarcoma specimens, respectively. Taken together, our results identify EphA2, a tyrosine kinase with known functions in neovascularization and oncogenesis, as an entry receptor for KSHV.
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Acknowledgements
This work was supported by the Akademie der Wissenschaften und der Literatur (Mainz), the European Community research project TargetHerpes, Deutsche Forschungsgemeinschaft center grant 643 and graduate school grant 1071 “viruses of the immune system.” M. Stürzl, E.N. and F.N. were supported by the interdisciplinary center for clinical research at the University Hospital of the University of Erlangen. M. Stürzl and E.N. were supported by the German Cancer Aid. We thank L. Wall for experimental assistance and R.C. Desrosiers for helpful discussions and critical reading of the manuscript. KSImm cells were kindly provided by A. Albini (Science and Technology Pole, IRCCS MultiMedica). 4G10 mouse hybridoma supernatant was kindly provided by B. Biesinger (Virologisches Institut, Universitätsklinikum Erlangen).
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A.S.H. and F.N. designed the study. A.S.H., J.K.K. and E.W. performed the key experiments. J.P.-I., K.S., M. Schmidt and A.H. performed real-time PCR experiments, cell culture and infection assays. S.K. performed mass spectrometry and analyzed the data. J.C., A.E., J.M. and N.H.B. contributed key reagents. E.N. and M. Stürzl helped with endothelial cell cultures and performed immunohistochemistry experiments. A.S.H. and F.N. wrote the manuscript. B.F. contributed expertise and helped write the paper.
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A. Hahn filed a patent to the United States Patent and Trademark Office involving the inhibition of EphA2 and KSHV infection with a small molecule.
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Hahn, A., Kaufmann, J., Wies, E. et al. The ephrin receptor tyrosine kinase A2 is a cellular receptor for Kaposi's sarcoma–associated herpesvirus. Nat Med 18, 961–966 (2012). https://doi.org/10.1038/nm.2805
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DOI: https://doi.org/10.1038/nm.2805
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