Camelids Nanobodies: Novel Prophylactic and Therapeutic Medication Against Epstein-Barr Virus Infection by Targeting Glycoprotein B
DOI:
https://doi.org/10.14738/aivp.1303.18942Keywords:
Epstein-Barr Virus (EBV), Glycoprotein B (gB), Camelids, Nanobodies, VHH, Antiviral Therapy, ProphylaxisAbstract
The Epstein-Barr virus (EBV) represents a widespread pathogen that infects nearly 98% of adults worldwide, creating persistent latent infections throughout the host's lifetime while contributing to various pathological conditions, encompassing malignancies and autoimmune diseases. EBV's presence within peripheral blood lymphocytes enables transmission through blood transfusion procedures and organ transplantation, creating substantial hazards, especially for immunocompromised patients including transplant recipients who face elevated vulnerability to EBV-associated lymphoproliferative conditions. Presently, neither efficacious vaccination programs nor specific antiviral treatments for EBV are available, highlighting the critical requirement for innovative preventive and treatment approaches. Glycoprotein B (gB) serves as a vital viral fusion protein, performing essential functions in EBV cellular entry, intercellular transmission, viral particle development, and nuclear exodus. Significantly, EBV particles containing elevated gB levels demonstrate superior infectious capacity, with gB establishing direct binding interactions with neuropilin-1 (NRP1) throughout epithelial cell and B lymphocyte invasion processes. Earlier studies demonstrate that EBV-gB successfully stimulates neutralizing antibody responses, establishing its potential as an optimal therapeutic target. This analysis suggests creating camelid-derived single-domain antibody components (VHHs), termed nanobodies (Nbs), directed against EBV-gB. These Nbs, measuring approximately 15 kDa, demonstrate substantially reduced size compared to traditional antibodies, being devoid of light chains and CH1 domains. Their distinctive characteristics encompass simplified production processes, exceptional stability, superior solubility, improved tissue and tumor infiltration capabilities, plus blood-brain barrier crossing potential. Importantly, Nbs demonstrate minimal immunogenic properties, reducing unwanted reactions. Large-volume Nb manufacturing could deliver easily administered prophylaxis for high-risk populations or therapeutic intervention for active infections. Camelid immunization using EBV-gB would simultaneously enable production of powerful neutralizing Nbs while permitting assessment of gB's vaccine potential. The extraordinary properties of Nbs constitute remarkable progress in therapeutic innovation, and we recommend focused initiatives to transform this promise into effective EBV treatments.
References
Arbabi-Ghahroudi, M. (2017). Camelid single-domain antibodies: Historical perspective and future outlook. Frontiers in Immunology, 8, 1589.
Arbabi-Ghahroudi, M. (2022). Camelid single-domain antibodies: promises and challenges as lifesaving treatments. International journal of molecular sciences, 23(9), 5009.
Arbabi-Ghahroudi, M., Desmyter, A., Wyns, L., Hamers, R., & Muyldermans, S. (1997). Selection and identification of single domain antibody fragments from camel heavy-chain antibodies. FEBS Letters, 414(3), 521–526.
Babcock, G. J., Decker, L. L., Volk, M., & Thorley-Lawson, D. A. (1998). EBV persistence in memory B cells in vivo. Immunity, 9(3), 395–404.
Bastawecy, I. M., Abdelmonem, M., Albert, E. L., & Abdullah, C. A. C. (2023). Epstein-Barr virus genetics: from
Bastawecy, I. M., Zaki, A. A., & Sobhy, N. M. (2014). Ovine herpesvirus 2 infection causes sudden death and abortion in susceptible animals. Life Science Journal, 11(10), 957-964.
Bobkov, V., Zarca, A. M., Van Hout, A., Arimont, M., Doijen, J., Bialkowska, M., ... & Smit, M. J. (2018). Nanobody-Fc constructs targeting chemokine receptor CXCR4 potently inhibit signaling and CXCR4-mediated HIV-entry and induce antibody effector functions. Biochemical pharmacology, 158, 413-424.
Brentville, V. A., Vankemmelbeke, M., Methingham, R. L., Symonds, P., Cook, K. W., Urbanowicz, R. A., ... & Durrant, L. G. (2021). A novel bivalent DNA vaccine encoding both spike protein receptor-binding domain and nucleocapsid protein of SARS-CoV-2 to elicit T cell and neutralising antibody responses that cross react with variants.
Bu, W., Joyce, M. G., Nguyen, H., Banh, D. V., Aguilar, F., Tariq, Z., ... & Cohen, J. I. (2019). Immunization with components of the viral fusion apparatus elicits antibodies that neutralize Epstein-Barr virus in B cells and epithelial cells. Immunity, 50(5), 1305-1316.
Burns, D. M., & Chaganti, S. (2021). Epstein-Barr virus-associated lymphoproliferative disorders in immunosuppressed patients. Annals of Lymphoma, 5.
Chen, W. H., Hajduczki, A., Martinez, E. J., Bai, H., Matz, H., Hill, T. M., ... & Joyce, M. G. (2023). Shark nanobodies with potent SARS-CoV-2 neutralizing activity and broad sarbecovirus reactivity. Nature Communications, 14(1),
Chi, X., Zhang, X., Pan, S., Yu, Y., Shi, Y., Lin, T., ... & Yang, W. (2022). An ultrapotent RBD-targeted biparatopic nanobody neutralizes broad SARS-CoV-2 variants. Signal transduction and targeted therapy, 7(1), 44.
Cohen, J. I. (2000). Epstein–Barr virus infection. New England journal of medicine, 343(7), 481-492.
Crauwels, M., Van Vaerenbergh, N., Kulaya, N. B., Vincke, C., D’Huyvetter, M., Devoogdt, N., ... & Xavier, C. (2020). Reshaping nanobodies for affinity purification on protein a. New Biotechnology, 57, 20-28.
Cui, X., Cao, Z., Ishikawa, Y., Cui, S., Imadome, K. I., & Snapper, C. M. (2021). Immunization with Epstein–Barr virus core fusion machinery envelope proteins elicit high titers of neutralizing activities and protect humanized mice from lethal dose EBV challenge. Vaccines, 9(3), 285.
De Beer, M. A., & Giepmans, B. N. (2020). Nanobody-based probes for subcellular protein identification and visualization. Frontiers in cellular neuroscience, 14, 573278.
De Genst, E., Saerens, D., Muyldermans, S., & Conrath, K. (2006). Antibody repertoire development in camelids. Developmental & Comparative Immunology, 30(1-2), 187-198.
Escalante, G. M., Foley, J., Mutsvunguma, L. Z., Rodriguez, E., Mulama, D. H., Muniraju, M., ... & Ogembo, J. G. (2020). A pentavalent Epstein-Barr virus-like particle vaccine elicits high titers of neutralizing antibodies against Epstein-Barr virus infection in immunized rabbits. Vaccines, 8(2), 169.
Escalante, G. M., Mutsvunguma, L. Z., Muniraju, M., Rodriguez, E., & Ogembo, J. G. (2022). Four decades of prophylactic EBV vaccine research: a systematic review and historical perspective. Frontiers in Immunology, 13,
Garcia, N. J., Chen, J., & Longnecker, R. (2013). Modulation of Epstein-Barr virus glycoprotein B (gB) fusion activity by the gB cytoplasmic tail domain. MBio, 4(1), 10-1128.
Hamers-Casterman, C. T. S. G., Atarhouch, T., Muyldermans, S. A., Robinson, G., Hammers, C., Songa, E. B., ... & Hammers, R. (1993). Naturally occurring antibodies devoid of light chains. Nature, 363(6428), 446-448.
Bell, A., & Pond, W. G. (2004). Encyclopedia of animal science. CRC Press.
He, S., Wang, J., Chen, H., Qian, Z., Hu, K., Shi, B., & Wang, J. (2023). A competitive panning method reveals an anti-SARS-CoV-2 nanobody specific for an RBD-ACE2 binding site. Vaccines, 11(2), 371.
Hong, J., Wei, D., Zhong, L., Wu, Q., Chen, K., Zhang, W., ... & Chen, Y. (2022). Glycoprotein B antibodies completely neutralize EBV infection of B Cells. Frontiers in Immunology, 13, 920467.
Hu, B., Liu, T., Li, L., Shi, L., Yao, M., Li, C., ... & Wang, F. (2022). IgG-binding nanobody capable of prolonging nanobody-based radiotracer plasma half-life and enhancing the efficacy of tumor-targeted radionuclide therapy. Bioconjugate chemistry, 33(7), 1328-1339.
Indari, O., Ghosh, S., Bal, A. S., James, A., Garg, M., Mishra, A., ... & Jha, H. C. (2024). Awakening the sleeping giant: Epstein–Barr virus reactivation by biological agents. Pathogens and Disease, 82, ftae002. IARC Working Group on the Evaluation of Carcinogenic Risks to Humans, International Agency for Research on
Cancer, & World Health Organization. (1997). Epstein-Barr Virus and Kaposi's Sarcoma Herpesvirus/human Herpesvirus 8: IARC Monograph on the Evaluation of Carcinogenic Risks to Humans. World Health Organization
Jean-Pierre, V., Lupo, J., Buisson, M., Morand, P., & Germi, R. (2021). Main targets of interest for the development of a prophylactic or therapeutic Epstein-Barr virus vaccine. Frontiers in microbiology, 12, 701611.Jin, B., Ye, Z.
Jin, B. K., Odongo, S., Radwanska, M., & Magez, S. (2023). NANOBODIES®: a review of diagnostic and therapeutic applications. International Journal of Molecular Sciences, 24(6), 5994-5994.
Jovčevska, I., & Muyldermans, S. (2020). The therapeutic potential of nanobodies. BioDrugs, 34(1), 11-26.
Khodabakhsh, F., Behdani, M., Rami, A., & Kazemi-Lomedasht, F. (2018). Single-domain antibodies or nanobodies: a class of next-generation antibodies. International reviews of immunology, 37(6), 316-322.Lauwereys, M., Ghahroudi, M. A., Desmyter, A., Kinne, J., Hölzer, W., De Genst, E., Wyns, L., & Muyldermans, S. (1998). Potent enzyme inhibitors derived from dromedary heavy-chain antibodies. The EMBO Journal, 17(13), 3512–3520.
Liu, B., & Yang, D. (2022). Easily established and multifunctional synthetic nanobody libraries as research tools. International Journal of Molecular Sciences, 23(3), 1482.
Mei, Y., Chen, Y., Sivaccumar, J. P., An, Z., Xia, N., & Luo, W. (2022). Research progress and applications of nanobody in human infectious diseases. Frontiers in pharmacology, 13, 963978.
Mitchell, L. S., & Colwell, L. J. (2018). Analysis of nanobody paratopes reveals greater diversity than classical antibodies. Protein Engineering, Design and Selection, 31(7-8), 267-275.
Mitchell, L. S., & Colwell, L. J. (2018). Comparative analysis of nanobody sequence and structure data. Proteins: Structure, Function, and Bioinformatics, 86(7), 697-706.
Morrison, C. (2019). Nanobody approval gives domain antibodies a boost. Nat Rev Drug Discov, 18(7), 485-487.
Mostafa, M. I., & Mohamed, A. (2023). Revolutionizing antiviral therapy with nanobodies: Generation and prospects. Archives of Virology, 168(3), 109.
Moutel, S., Bery, N., Bernard, V., Keller, L., Lemesre, E., De Marco, A., ... & Perez, F. (2016). NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies. Elife, 5, e16228.
Münz, C. (2019). Latency and lytic replication in Epstein–Barr virus-associated oncogenesis. Nature Reviews Microbiology, 17(11), 691-700.
Murata, T., & Tsurumi, T. (2014). Switching of EBV cycles between latent and lytic states: Regulation of EBV reactivation. Reviews in Medical Virology, 24(3), 142–153.
Muyldermans, S. (2013). Nanobodies: natural single-domain antibodies. Annual review of biochemistry, 82(1), 775-797.
Muyldermans, S. (2021). A guide to: generation and design of nanobodies. The FEBS journal, 288(7), 2084-2102.
Muyldermans, S. (2021). Applications of nanobodies. Annual review of animal biosciences, 9(1), 401-421.
Muyldermans, S., Cambillau, C., & Wyns, L. (2001). Recognition of antigens by single-domain antibody fragments: the superfluous luxury of paired domains. Trends in biochemical sciences, 26(4), 230-235.
Neuhierl, B., Feederle, R., Hammerschmidt, W., & Delecluse, H. J. (2002). Glycoprotein gp110 of Epstein–Barr virus determines viral tropism and efficiency of infection. Proceedings of the National Academy of Sciences, 99(23), 15036-15041.
Papesch, M., & Watkins, R. (2001). Epstein‐Barr virus infectious mononucleosis. Clinical Otolaryngology & Allied Sciences, 26(1), 3-8.
Pellett, P. E., Kousoulas, K. G., Pereira, L., & Roizman, B. (1985). Anatomy of the herpes simplex virus 1 strain F glycoprotein B gene: primary sequence and predicted protein structure of the wild type and of monoclonal antibody-resistant mutants. Journal of Virology, 53(1), 243-253.
Plotkin, S. A., Wang, D., Oualim, A., Diamond, D. J., Kotton, C. N., Mossman, S., ... & Dormitzer, P. R. (2020). The status of vaccine development against the human cytomegalovirus. The Journal of infectious diseases, 221(Supplement_1), S113-S122.
Rahbarizadeh, F., Rasaee, M. J., Forouzandeh, M., Allameh, A., Sarrami, R., Nasiry, H., & Sadeghizadeh, M. (2005). The production and characterization of novel heavy-chain antibodies against the tandem repeat region of MUC1 mucin. Immunological investigations, 34(4), 431-452.
Reimer, J. J., Backovic, M., Deshpande, C. G., Jardetzky, T., & Longnecker, R. (2009). Analysis of Epstein-Barr virus glycoprotein B functional domains via linker insertion mutagenesis. Journal of virology, 83(2), 734-747.
Richards, D. A. (2018). Exploring alternative antibody scaffolds: Antibody fragments and antibody mimics for targeted drug delivery. Drug Discovery Today: Technologies, 30, 35-46.
Kieff, E. (2007). Epstin-Barr virus and its replication. Fields virology, 2, 2603-2654.
Ruiz-López, E., & Schuhmacher, A. J. (2021). Transportation of single-domain antibodies through the blood–brain barrier. Biomolecules, 11(8), 1131.
Salvador, J. P., Vilaplana, L., & Marco, M. P. (2019). Nanobody: outstanding features for diagnostic and therapeutic applications. Analytical and bioanalytical chemistry, 411, 1703-1713.
Scarpini, S., Morigi, F., Betti, L., Dondi, A., Biagi, C., & Lanari, M. (2021). Development of a vaccine against human cytomegalovirus: advances, barriers, and implications for the clinical practice. Vaccines, 9(6), 551.
Schleiss, M. R., Permar, S. R., & Plotkin, S. A. (2017). Progress toward development of a vaccine against congenital cytomegalovirus infection. Clinical and Vaccine Immunology, 24(12), e00268-17.
Sehrawat, S., & Singh, A. (2006). Anti-erythrocyte natural antibody activity in the unconventional ‘heavy chain’immunoglobulins of Indian desert camel (Camelus dromedarius). Veterinary immunology and immunopathology, 111(3-4), 139-147.
Smatti, M. K., Al-Sadeq, D. W., Ali, N. H., Pintus, G., Abou-Saleh, H., & Nasrallah, G. K. (2018). Epstein–Barr virus epidemiology, serology, and genetic variability of LMP-1 oncogene among healthy population: an update. Frontiers in oncology, 8, 211.
Smatti, M. K., Yassine, H. M., AbuOdeh, R., AlMarawani, A., Taleb, S. A., Althani, A. A., & Nasrallah, G. K. (2017). Prevalence and molecular profiling of Epstein Barr virus (EBV) among healthy blood donors from different nationalities in Qatar. PLoS One, 12(12), e0189033.
Sockolosky, J. T., Dougan, M., Ingram, J. R., Ho, C. C. M., Kauke, M. J., Almo, S. C., ... & Garcia, K. C. (2016). Durable antitumor responses to CD47 blockade require adaptive immune stimulation. Proceedings of the National Academy of Sciences, 113(19), E2646-E2654.
Sun, C., Chen, X. C., Kang, Y. F., & Zeng, M. S. (2021). The status and prospects of Epstein–Barr virus prophylactic vaccine development. Frontiers in Immunology, 12, 677027.
Valdés-Tresanco, M. S., Molina-Zapata, A., Pose, A. G., & Moreno, E. (2022). Structural insights into the design of synthetic nanobody libraries. Molecules, 27(7), 2198.
Vincke, C., Loris, R., Saerens, D., Martinez-Rodriguez, S., Muyldermans, S., & Conrath, K. (2009). General strategy to humanize a camelid single-domain antibody and identification of a universal humanized nanobody scaffold. Journal of Biological Chemistry, 284(5), 3273-3284.
Vu, K. B., Ghahroudi, M. A., Wyns, L., & Muyldermans, S. (1997). Comparison of llama VH sequences from conventional and heavy chain antibodies. Molecular immunology, 34(16-17), 1121-1131.
Wagner, H. J., Wehrle, S., Weiss, E., Cavallari, M., & Weber, W. (2018). A two-step approach for the design and generation of nanobodies. International journal of molecular sciences, 19(11), 3444.
Wahner-Roedler, D. L., & Kyle, R. A. (2005). Heavy chain diseases. Best Practice & Research Clinical Haematology, 18(4), 729–746.
Wang, H. B., Zhang, H., Zhang, J. P., Li, Y., Zhao, B., Feng, G. K., ... & Zeng, M. S. (2015). Neuropilin 1 is an entry factor that promotes EBV infection of nasopharyngeal epithelial cells. Nature communications, 6(1), 6240.
Wolf, H. A. N. S., Haus, M. A. R. G. R. E. T., & Wilmes, E. B. E. R. H. A. R. D. (1984). Persistence of Epstein-Barr virus in the parotid gland. Journal of virology, 51(3), 795-798.
Wong, Y., Meehan, M. T., Burrows, S. R., Doolan, D. L., & Miles, J. J. (2022). Estimating the global burden of Epstein–Barr virus-r Wrapp, D., De Vlieger, D., Corbett, K. S., Torres, G. M., Wang, N., Van Breedam, W., ... & McLellan, J. S. (2020). Structural basis for potent neutralization of betacoronaviruses by single-domain camelid antibodies. Cell, 181(5), 1004-1015.
Yang, E. Y., & Shah, K. (2020). Nanobodies: next generation of cancer diagnostics and therapeutics. Frontiers in Oncology, 10, 1182.
Ye, J., Wei, J., Luo, Y., Deng, Y., Que, T., Zhang, X., ... & Luo, X. (2021). Epstein-barr virus promotes tumor angiogenesis by activating STIM1-dependent Ca2+ signaling in nasopharyngeal carcinoma. Pathogens, 10(10), Yokoe, S., Hasuike, A., Watanabe, N., Tanaka, H., Karahashi, H., Wakuda, S., ... & Imai, K. (2022). Epstein-Barr virus promotes the production of inflammatory cytokines in gingival fibroblasts and RANKL-induced osteoclast differentiation in RAW264. 7 cells. International journal of molecular sciences, 23(2), 809.
Zambrano, N., Froechlich, G., Lazarevic, D., Passariello, M., Nicosia, A., De Lorenzo, C., ... & Sasso, E. (2022). Highthroughput monoclonal antibody discovery from phage libraries: challenging the current preclinical pipeline to keep the pace with the increasing mAb demand. Cancers, 14(5), 1325.
Zavrtanik, U., Lukan, J., Loris, R., Lah, J., & Hadži, S. (2018). Structural basis of epitope recognition by heavy-chain camelid antibodies. Journal of molecular biology, 430(21), 4369-4386.
Zhao, G. X., Fang, X. Y., Bu, G. L., Chen, S. J. B., Sun, C., Li, T., ... & Zeng, M. S. (2024). Potent human monoclonal antibodies targeting Epstein-Barr virus gp42 reveal vulnerable sites for virus infection. Cell Reports Medicine, 5(5).
Zhu, Q. Y., Kong, X. W., Sun, C., Xie, S. H., Hildesheim, A., Cao, S. M., & Zeng, M. S. (2020). Association between antibody responses to Epstein-Barr virus glycoproteins, neutralization of infectivity, and the risk of nasopharyngeal carcinoma. Msphere, 5(6), 10-1128.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Iman M. Bastawecy, Ahmed F. Affy, Mohamed Abdelmonem, Fatima-Alzahraa Gamal, Maha A. Alhadad
This work is licensed under a Creative Commons Attribution 4.0 International License.
