Antigenic and structural insights into Langya henipavirus attachment glycoprotein

Yaohui Li; Xiaoyan Huang; Xiaodong Zai; Chenfeng Mao; Ruihua Li; Yamei Feng; Yue Zhang; Zhang Zhang; Jun Zhang; Junjie Xu

doi:10.1016/j.virs.2025.08.005

October 2025

Yaohui Li, Xiaoyan Huang, Xiaodong Zai, Chenfeng Mao, Ruihua Li, Yamei Feng, Yue Zhang, Zhang Zhang, Jun Zhang and Junjie Xu. Antigenic and structural insights into Langya henipavirus attachment glycoprotein[J]. Virologica Sinica, 2025, 40(5): 769-777. doi: 10.1016/j.virs.2025.08.005

Citation: Yaohui Li, Xiaoyan Huang, Xiaodong Zai, Chenfeng Mao, Ruihua Li, Yamei Feng, Yue Zhang, Zhang Zhang, Jun Zhang, Junjie Xu. Antigenic and structural insights into Langya henipavirus attachment glycoprotein .VIROLOGICA SINICA, 2025, 40(5) : 769-777. http://dx.doi.org/10.1016/j.virs.2025.08.005

琅琊亨尼帕病毒附着糖蛋白的抗原和结构研究

北京生物工程研究所前沿生物技术实验室

通讯作者： 徐俊杰, xujunjie@sina.com
收稿日期： 2024-12-31
录用日期： 2025-08-29

摘要

亨尼帕病毒属是一类单股负链RNA病毒，其中尼帕病毒（NiV）和亨德拉病毒（HeV）可导致人畜共患的严重疾病。2022年在中国发现的新型亨尼帕病毒——琅琊病毒（LayV）引起了广泛关注。本文解析了LayV附着糖蛋白（G蛋白）的晶体结构（3.4 Å分辨率），结果显示该蛋白头部区域由六个类似β-螺旋桨的结构域构成，并具有区别于NiV和HeV的独特构象及糖基化修饰特征。受体结合位点的空间位阻导致LayV G蛋白无法与Ephrin-B2/B3受体结合，同时也不与现有亨尼帕病毒中和抗体结合。免疫原性研究表明，LayV G蛋白在小鼠中可诱导针对LayV和墨江病毒（MojV）的强抗体应答，但与其他亨尼帕病毒的交叉反应性较弱。进一步地，本研究获得了9株针对LayV G蛋白的单克隆抗体，其中2株表现出对不同进化支系亨尼帕病毒G蛋白的广谱结合活性。本研究揭示了LayV G蛋白独特的结构与抗原特性，为琅琊病毒等新型亨尼帕病毒的疫苗和抗体研发提供了重要参考。
- 琅琊病毒
- , 亨尼帕病毒
- , 附着糖蛋白
- , 晶体结构
- , 糖基化
- , 免疫原性
- , 单克隆抗体

Antigenic and structural insights into Langya henipavirus attachment glycoprotein

Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, 100071, China

Corresponding author: Junjie Xu, xujunjie@sina.com
Received Date: 31 December 2024
Accepted Date: 29 August 2025

Abstract

The invasion of host cells by the henipavirus is facilitated through the interaction between viral attachment (G) and fusion (F) glycoproteins with receptors on the cell surface. Langya henipavirus (LayV) was newly identified in China in 2022. The G proteins of LayV and Mojiang virus (MojV) exhibit high amino acid homology (86%), while they are located in a unique evolutionary clade within the Henipavirus genus. In this study, the crystal structure of the LayV G protein was resolved at a 3.37 Å resolution, revealing a head domain with six β-propeller-like domains distinct from other henipavirus G proteins, such as those of Nipah virus (NiV) and Hendra virus (HeV). Furthermore, the prominent loop in the center cavity of the LayV G protein showed unique structural features. In the ELISA and SPR assays, the LayV G protein was unable to bind to the existing henipavirus-neutralizing antibodies or the ephrin-B2 receptor. Immunogenicity studies in mice demonstrated robust antibody responses elicited by the LayV G protein. These antibodies exhibited strong reactivity against both LayV and MojV G proteins. However, only weak cross-reactivity was observed with other henipaviruses. Moreover, eight monoclonal antibodies targeting the LayV G protein were generated, two of which exhibited broad binding activity across different henipavirus G proteins. These findings underscore the need for tailored vaccines and therapeutics for LayV and related novel henipaviruses
- Langya henipavirus (LayV)
- , Attachment glycoprotein
- , Crystal structure
- , Glycosylation
- , Immunogenicity
- , Monoclonal antibodies

References
1. Avanzato, V.A., Oguntuyo, K.Y., Escalera-Zamudio, M., Gutierrez, B., Golden, M., Kosakovsky Pond, S.L., Pryce, R., Walter, T.S., Seow, J., Doores, K.J., Pybus, O.G., Munster, V.J., Lee, B.,Bowden, T.A., 2019. A structural basis for antibody-mediated neutralization of Nipah virus reveals a site of vulnerability at the fusion glycoprotein apex. Proc Natl Acad Sci U S A, 116, 25057-25067.
2. Bonaparte, M.I., Dimitrov, A.S., Bossart, K.N., Crameri, G., Mungall, B.A., Bishop, K.A., Choudhry, V., Dimitrov, D.S., Wang, L.F., Eaton, B.T.,Broder, C.C., 2005. Ephrin-B2 ligand is a functional receptor for Hendra virus and Nipah virus. Proc Natl Acad Sci U S A, 102, 10652-10657.
3. Bowden, T.A., Aricescu, A.R., Gilbert, R.J., Grimes, J.M., Jones, E.Y.,Stuart, D.I., 2008a. Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. Nat Struct Mol Biol, 15, 567-572.
4. Bowden, T.A., Crispin, M., Harvey, D.J., Aricescu, A.R., Grimes, J.M., Jones, E.Y.,Stuart, D.I., 2008b. Crystal structure and carbohydrate analysis of Nipah virus attachment glycoprotein: a template for antiviral and vaccine design. J Virol, 82, 11628-11636.
5. Bradel-Tretheway, B.G., Zamora, J.L.R., Stone, J.A., Liu, Q., Li, J.,Aguilar, H.C., 2019. Nipah and Hendra Virus Glycoproteins Induce Comparable Homologous but Distinct Heterologous Fusion Phenotypes. J Virol, 93, e00577-19.
6. Byrne, P.O., Fisher, B.E., Ambrozak, D.R., Blade, E.G., Tsybovsky, Y., Graham, B.S., Mclellan, J.S.,Loomis, R.J., 2023. Structural basis for antibody recognition of vulnerable epitopes on Nipah virus F protein. Nat Commun, 14, 1494.
7. Byrne, P.O., Blade, E.G., Fisher, B.E., Ambrozak, D.R., Ramamohan, A.R., Graham, B.S., Loomis, R.J.,Mclellan, J.S., 2024. Prefusion stabilization of the Hendra and Langya virus F proteins. J Virol, 98, e0137223.
8. Centers for Disease Control and Prevention (CDC).1999. Outbreak of Hendra-like virus--Malaysia and Singapore, 1998-1999. Can Commun Dis Rep, 25, 108-112.
9. Cheliout Da Silva, S., Yan, L., Dang, H.V., Xu, K., Epstein, J.H., Veesler, D.,Broder, C.C., 2021. Functional Analysis of the Fusion and Attachment Glycoproteins of Mojiang Henipavirus. Viruses, 13, 517.
10. Clayton, B.A., Middleton, D., Arkinstall, R., Frazer, L., Wang, L.F.,Marsh, G.A., 2016. The Nature of Exposure Drives Transmission of Nipah Viruses from Malaysia and Bangladesh in Ferrets. PLoS Negl Trop Dis, 10, e0004775.
11. Dang, H.V., Chan, Y.P., Park, Y.J., Snijder, J., Da Silva, S.C., Vu, B., Yan, L., Feng, Y.R., Rockx, B., Geisbert, T.W., Mire, C.E., Broder, C.C.,Veesler, D., 2019. An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections. Nat Struct Mol Biol, 26, 980-987.
12. Dang, H.V., Cross, R.W., Borisevich, V., Bornholdt, Z.A., West, B.R., Chan, Y.P., Mire, C.E., Da Silva, S.C., Dimitrov, A.S., Yan, L., Amaya, M., Navaratnarajah, C.K., Zeitlin, L., Geisbert, T.W., Broder, C.C.,Veesler, D., 2021. Broadly neutralizing antibody cocktails targeting Nipah virus and Hendra virus fusion glycoproteins. Nat Struct Mol Biol, 28, 426-434.
13. Dong, J., Cross, R.W., Doyle, M.P., Kose, N., Mousa, J.J., Annand, E.J., Borisevich, V., Agans, K.N., Sutton, R., Nargi, R., Majedi, M., Fenton, K.A., Reichard, W., Bombardi, R.G., Geisbert, T.W.,Crowe, J.E., Jr., 2020. Potent Henipavirus Neutralization by Antibodies Recognizing Diverse Sites on Hendra and Nipah Virus Receptor Binding Protein. Cell, 183, 1536-1550 e1517.
14. Ferruz, N., Schmidt, S.,Hocker, B., 2021. ProteinTools: a toolkit to analyze protein structures. Nucleic Acids Res, 49, W559-W566.
15. Gomez Roman, R., Tornieporth, N., Cherian, N.G., Shurtleff, A.C., L'azou Jackson, M., Yeskey, D., Hacker, A., Mungai, E.,Le, T.T., 2022. Medical countermeasures against henipaviruses: a review and public health perspective. Lancet Infect Dis, 22, e13-e27.
16. Guo, Y., Wu, S., Li, W., Yang, H., Shi, T., Ju, B., Zhang, Z.,Yan, R., 2024. The cryo-EM structure of homotetrameric attachment glycoprotein from langya henipavirus. Nat Commun, 15, 812.
17. Isaacs, A., Low, Y.S., Macauslane, K.L., Seitanidou, J., Pegg, C.L., Cheung, S.T.M., Liang, B., Scott, C.a.P., Landsberg, M.J., Schulz, B.L., Chappell, K.J., Modhiran, N.,Watterson, D., 2023. Structure and antigenicity of divergent Henipavirus fusion glycoproteins. Nat Commun, 14, 3577.
18. Laing, E.D., Navaratnarajah, C.K., Cheliout Da Silva, S., Petzing, S.R., Xu, Y., Sterling, S.L., Marsh, G.A., Wang, L.F., Amaya, M., Nikolov, D.B., Cattaneo, R., Broder, C.C.,Xu, K., 2019. Structural and functional analyses reveal promiscuous and species specific use of ephrin receptors by Cedar virus. Proc Natl Acad Sci U S A, 116, 20707-20715.
19. Lee, B., Pernet, O., Ahmed, A.A., Zeltina, A., Beaty, S.M.,Bowden, T.A., 2015. Molecular recognition of human ephrinB2 cell surface receptor by an emergent African henipavirus. Proc Natl Acad Sci U S A, 112, E2156-2165.
20. Lee, S.H., Kim, K., Kim, J., No, J.S., Park, K., Budhathoki, S., Lee, S.H., Lee, J., Cho, S.H., Cho, S., Lee, G.Y., Hwang, J., Kim, H.C., Klein, T.A., Uhm, C.S., Kim, W.K.,Song, J.W., 2021. Discovery and Genetic Characterization of Novel Paramyxoviruses Related to the Genus Henipavirus in Crocidura Species in the Republic of Korea. Viruses, 13, 2020.
21. Li, Y., Li, R., Wang, M., Liu, Y., Yin, Y., Zai, X., Song, X., Chen, Y., Xu, J.,Chen, W., 2020. Fc-Based Recombinant Henipavirus Vaccines Elicit Broad Neutralizing Antibody Responses in Mice. Viruses, 12, 480.
22. Madera, S., Kistler, A., Ranaivoson, H.C., Ahyong, V., Andrianiaina, A., Andry, S., Raharinosy, V., Randriambolamanantsoa, T.H., Ravelomanantsoa, N.a.F., Tato, C.M., Derisi, J.L., Aguilar, H.C., Lacoste, V., Dussart, P., Heraud, J.M.,Brook, C.E., 2022. Discovery and Genomic Characterization of a Novel Henipavirus, Angavokely Virus, from Fruit Bats in Madagascar. J Virol, 96, e0092122.
23. Marsh, G.A., De Jong, C., Barr, J.A., Tachedjian, M., Smith, C., Middleton, D., Yu, M., Todd, S., Foord, A.J., Haring, V., Payne, J., Robinson, R., Broz, I., Crameri, G., Field, H.E.,Wang, L.F., 2012. Cedar virus: a novel Henipavirus isolated from Australian bats. PLoS Pathog, 8, e1002836.
24. May Aaron, J., Pothula Karunakar, R., Janowska, K.,Acharya, P., 2023. Structures of Langya Virus Fusion Protein Ectodomain in Pre- and Postfusion Conformation. Journal of Virology, 97, e00433-00423.
25. Mire, C.E., Chan, Y.P., Borisevich, V., Cross, R.W., Yan, L., Agans, K.N., Dang, H.V., Veesler, D., Fenton, K.A., Geisbert, T.W.,Broder, C.C., 2020. A Cross-Reactive Humanized Monoclonal Antibody Targeting Fusion Glycoprotein Function Protects Ferrets Against Lethal Nipah Virus and Hendra Virus Infection. J Infect Dis, 221, S471-S479.
26. Negrete, O.A., Wolf, M.C., Aguilar, H.C., Enterlein, S., Wang, W., Muhlberger, E., Su, S.V., Bertolotti-Ciarlet, A., Flick, R.,Lee, B., 2006. Two key residues in ephrinB3 are critical for its use as an alternative receptor for Nipah virus. PLoS Pathog, 2, e7.
27. Rissanen, I., Ahmed, A.A., Azarm, K., Beaty, S., Hong, P., Nambulli, S., Duprex, W.P., Lee, B.,Bowden, T.A., 2017. Idiosyncratic Mojiang virus attachment glycoprotein directs a host-cell entry pathway distinct from genetically related henipaviruses. Nat Commun, 8, 16060.
28. Taylor, J., Thompson, K., Annand, E.J., Massey, P.D., Bennett, J., Eden, J.S., Horsburgh, B.A., Hodgson, E., Wood, K., Kerr, J., Kirkland, P., Finlaison, D., Peel, A.J., Eby, P.,Durrheim, D.N., 2022. Novel variant Hendra virus genotype 2 infection in a horse in the greater Newcastle region, New South Wales, Australia. One Health, 15, 100423.
29. Wang, Z., Amaya, M., Addetia, A., Dang, H.V., Reggiano, G., Yan, L., Hickey, A.C., Dimaio, F., Broder, C.C.,Veesler, D., 2022a. Architecture and antigenicity of the Nipah virus attachment glycoprotein. Science, 375, 1373-1378.
30. Wang, Z., Dang, H.V., Amaya, M., Xu, Y., Yin, R., Yan, L., Hickey, A.C., Annand, E.J., Horsburgh, B.A., Reid, P.A., Smith, I., Eden, J.S., Xu, K., Broder, C.C.,Veesler, D., 2022b. Potent monoclonal antibody-mediated neutralization of a divergent Hendra virus variant. Proc Natl Acad Sci U S A, 119, e2122769119.
31. Wang, C., Li, M., Wang, Y., Ding, Q., Fan, S.,Lan, J., 2024. Structural insights into the Langya virus attachment glycoprotein. Structure, 32, 1090-1098 e1093.
32. Wang, Z., Mccallum, M., Yan, L., Gibson, C.A., Sharkey, W., Park, Y.J., Dang, H.V., Amaya, M., Person, A.,Broder, C.C., 2024. Structure and design of Langya virus glycoprotein antigens. Proc Natl Acad Sci U S A., 121, e2314990121.
33. Williamson, M.M., Hooper, P.T., Selleck, P.W., Gleeson, L.J., Daniels, P.W., Westbury, H.A.,Murray, P.K., 1998. Transmission studies of Hendra virus (equine morbillivirus) in fruit bats, horses and cats. Aust Vet J, 76, 813-818.
34. Wu, Z., Yang, L., Yang, F., Ren, X., Jiang, J., Dong, J., Sun, L., Zhu, Y., Zhou, H.,Jin, Q., 2014. Novel Henipa-like virus, Mojiang Paramyxovirus, in rats, China, 2012. Emerg Infect Dis, 20, 1064-1066.
35. Xu, Y., Lou, Z., Liu, Y., Cole, D.K., Su, N., Qin, L., Li, X., Bai, Z., Rao, Z.,Gao, G.F., 2004. Crystallization and preliminary crystallographic analysis of the fusion core from two new zoonotic paramyxoviruses, Nipah virus and Hendra virus. Acta Crystallogr D Biol Crystallogr, 60, 1161-1164.
36. Xu, K., Rockx, B., Xie, Y., Debuysscher, B.L., Fusco, D.L., Zhu, Z., Chan, Y.P., Xu, Y., Luu, T., Cer, R.Z., Feldmann, H., Mokashi, V., Dimitrov, D.S., Bishop-Lilly, K.A., Broder, C.C.,Nikolov, D.B., 2013. Crystal structure of the Hendra virus attachment G glycoprotein bound to a potent cross-reactive neutralizing human monoclonal antibody. PLoS Pathog, 9, e1003684.
37. Zhang, X.A., Li, H., Jiang, F.C., Zhu, F., Zhang, Y.F., Chen, J.J., Tan, C.W., Anderson, D.E., Fan, H., Dong, L.Y., Li, C., Zhang, P.H., Li, Y., Ding, H., Fang, L.Q., Wang, L.F.,Liu, W., 2022. A Zoonotic Henipavirus in Febrile Patients in China. N Engl J Med, 387, 470-472.
38. Zhu, Z., Dimitrov, A.S., Bossart, K.N., Crameri, G., Bishop, K.A., Choudhry, V., Mungall, B.A., Feng, Y.R., Choudhary, A., Zhang, M.Y., Feng, Y., Wang, L.F., Xiao, X., Eaton, B.T., Broder, C.C.,Dimitrov, D.S., 2006. Potent neutralization of Hendra and Nipah viruses by human monoclonal antibodies. J Virol, 80, 891-899.
Proportional views
Electronic Supplementary Material

10.1016j.virs.2025.08.005-ESM1.docx