Quantification, epitope mapping and genotype cross-reactivity of hepatitis B preS-specific antibodies in subjects vaccinated with different dosage regimens of BM32
Inna Tulaeva, Carolin Cornelius, Petra Zieglmayer, René Zieglmayer, René Schmutz, Patrick Lemell, Milena Weber, Margarete Focke-Tejkl, Alexander Karaulov, Rainer Henning, Rudolf Valenta, Inna Tulaeva, Carolin Cornelius, Petra Zieglmayer, René Zieglmayer, René Schmutz, Patrick Lemell, Milena Weber, Margarete Focke-Tejkl, Alexander Karaulov, Rainer Henning, Rudolf Valenta
Abstract
Background: Chronic hepatitis B virus (HBV) infections are a global health problem. There is a need for therapeutic strategies blocking continuous infection of liver cells. The grass pollen allergy vaccine BM32 containing the preS domain of the large HBV surface protein (LHBs) as immunogenic carrier induced IgG antibodies in human subjects inhibiting HBV infection in vitro. Aim of this study was the quantification, epitope mapping and investigation of HBV genotype cross-reactivity of preS-specific antibodies in subjects treated with different dosage regimens of BM32 METHODS: Hundred twenty eight grass pollen allergic patients received in a double-blind, placebo-controlled trial five monthly injections of placebo (aluminum hydroxide, n= 34) or different courses of BM32 (2 placebo + 3 BM32, n= 33; 1 placebo + 4 BM32, n= 30; 5 BM32, n= 31). Recombinant Escherichia coli-expressed preS was purified. Overlapping peptides spanning preS and the receptor-binding sites from consensus sequences of genotypes A-H were synthesized and purified. Isotype (IgM, IgG, IgA, IgE) and IgG subclass (IgG1-IgG4) responses to preS and peptides were determined by ELISA at baseline, one and four months after the last injection. IgG1 and IgG4 subclass concentrations specific for preS and the receptor-binding site were measured by quantitative ELISA.
Findings: Five monthly injections induced the highest levels of preS-specific IgG consisting mainly of IgG1 and IgG4, with a sum of median preS-specific IgG1 and IgG4 concentrations of >135 μg/ml reaching up to 1.8 mg/ml. More than 20% of preS-specific IgG was directed against the receptor-binding site. BM32-induced IgG cross-reacted with the receptor-binding domains from all eight HBV genotypes A-H.
Interpretation: BM32 induces high levels of IgG1 and IgG4 antibodies against the receptor binding sites of all eight HBV genotypes and hence might be suitable for therapeutic HBV vaccination.
Funding: This study was supported by the PhD program IAI (KPW01212FW), by Viravaxx AG and by the Danube-ARC funded by the Government of Lower Austria. Rudolf Valenta is a recipient of a Megagrant of the Government of the Russian Federation, grant No 14.W03.31.0024.
Keywords: Antibody response; Epitopes; Genotype cross-reactivity; Grass pollen allergy vaccine BM32; Hepatitis B; Vaccination; preS.
Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.
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References
- Gerlich W.H. Medical virology of hepatitis B: how it began and where we are now. Virol J. 2013;10:1. doi: 10.1186/1743-422X-10-239.
- Revill P.A., Chisari F.V., Block J.M., Dandri M., Gehring A.J., Guo H. A global scientific strategy to cure hepatitis B. Lancet Gastroenterol Hepatol. 2019;4:545–558. doi: 10.1016/S2468-1253(19)30119-0.
- Gerlich W.H. Prophylactic vaccination against hepatitis B: achievements, challenges and perspectives. Med Microbiol Immunol. 2015;204:39–55. doi: 10.1007/s00430-014-0373-y.
- Neurath A.R., Kent S.B.H., Strick N., Parker K. Identification and chemical synthesis of a host cell receptor binding site on hepatitis B virus. Cell. 1986;46:429–436. doi: 10.1016/0092-8674(86)90663-X.
- Neurath A.R., Seto B., Strick N. Antibodies to synthetic peptides from the preS1 region of the hepatitis B virus (HBV) envelope (env) protein are virus-neutralizing and protective. Vaccine. 1989;7:234–236. doi: 10.1016/0264-410X(89)90235-1.
- Glebe D., Urban S., Knoop E.V., Çaǧ N., Krass P., Grün S. Mapping of the hepatitis B virus attachment site by use of infection-inhibiting preS1 lipopeptides and tupaia hepatocytes. Gastroenterology. 2005;129:234–245. doi: 10.1053/j.gastro.2005.03.090.
- Gripon P., Cannie I., Urban S. Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein. J Virol. 2005;79:1613–1622. doi: 10.1128/JVI.79.3.1613-1622.2005.
- Yan H., Zhong G., Xu G., He W., Jing Z., Gao Z. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. Elife. 2012;2012:1–28. doi: 10.7554/eLife.00049.
- Verrier E.R., Colpitts C.C., Sureau C., Baumert T.F. Hepatitis B virus receptors and molecular drug targets. Hepatol Int. 2016;10:567–573. doi: 10.1007/s12072-016-9718-5.
- Michel M.-.L., Pol S., Brechot C., Tiollais P. Immunotherapy of chronic hepatitis B by anti HBV vaccine: from present to future. Vaccine. 2001;19:2395–2399. doi: 10.1016/S0264-410X(00)00461-8.
- Shouval D., Roggendorf H., Roggendorf M. Enhanced immune response to hepatitis B vaccination through immunization with a Pre-S1/Pre-S2/S Vaccine. Med Microbiol Immunol. 2015;204:57–68. doi: 10.1007/s00430-014-0374-x.
- Valenta R., Campana R., Focke-Tejkl M., Niederberger V. Vaccine development for allergen-specific immunotherapy based on recombinant allergens and synthetic allergen peptides: lessons from the past and novel mechanisms of action for the future. J Allergy Clin Immunol. 2016;137:351–357. doi: 10.1016/j.jaci.2015.12.1299.
- Valenta R., Campana R., Niederberger V. Recombinant allergy vaccines based on allergen-derived B cell epitopes. Immunol Lett. 2017;189:19–26. doi: 10.1016/j.imlet.2017.04.015.
- Eckl-Dorna J., Weber M., Stanek V., Linhart B., Ristl R., Waltl E.E. Two years of treatment with the recombinant grass pollen allergy vaccine BM32 induces a continuously increasing allergen-specific IgG4 response. EBioMedicine. 2019;50:421–432. doi: 10.1016/j.ebiom.2019.11.006.
- Zieglmayer P., Focke-Tejkl M., Schmutz R., Lemell P., Zieglmayer R., Weber M. Mechanisms, safety and efficacy of a B cell epitope-based vaccine for immunotherapy of grass pollen allergy. EBioMedicine. 2016;11:43–57. doi: 10.1016/j.ebiom.2016.08.022.
- Niederberger V., Neubauer A., Gevaert P., Zidarn M., Worm M., Aberer W. Safety and efficacy of immunotherapy with the recombinant B-cell epitope–based grass pollen vaccine BM32. J Allergy Clin Immunol. 2018;142 doi: 10.1016/j.jaci.2017.09.052. 497-509.e9.
- Cornelius C., Schöneweis K., Georgi F., Weber M., Niederberger V., Zieglmayer P. Immunotherapy with the PreS-based grass pollen allergy vaccine BM32 induces antibody responses protecting against hepatitis B infection. EBioMedicine. 2016;11:58–67. doi: 10.1016/j.ebiom.2016.07.023.
- Niespodziana K., Focke-Tejkl M., Linhart B., Civaj V., Blatt K., Valent P. A hypoallergenic cat vaccine based on Fel d 1–derived peptides fused to hepatitis B PreS. J Allergy Clin Immunol. 2011;127 doi: 10.1016/j.jaci.2011.02.004. 1562-1570.e6.
- Paran N. HBV infection of cell culture: evidence for multivalent and cooperative attachment. EMBO J. 2001;20:4443–4453. doi: 10.1093/emboj/20.16.4443.
- Gallerano D., Ndlovu P., Makupe I., Focke-Tejkl M., Fauland K., Wollmann E. Comparison of the specificities of IgG, IgG-subclass, IgA and IgM reactivities in African and European HIV-infected individuals with an HIV-1 clade C proteome-based array. PLoS One. 2015;10:1–19. doi: 10.1371/journal.pone.0117204.
- Hayer J., Jadeau F., Deleage G., Kay A., Zoulim F., Combet C. HBVdb: a knowledge database for Hepatitis B Virus. Nucleic Acids Res. 2013;41:D566–D570. doi: 10.1093/nar/gks1022.
- Larsson A. AliView: a fast and lightweight alignment viewer and editor for large datasets. Bioinformatics. 2014;30:3276–3278. doi: 10.1093/bioinformatics/btu531.
- Kahila Bar-Gal G., Kim M.J., Klein A., Shin D.H., Oh C.S., Kim J.W. Tracing hepatitis B virus to the 16th century in a Korean mummy. Hepatology. 2012;56:1671–1680. doi: 10.1002/hep.25852.
- Patterson Ross Z., Klunk J., Fornaciari G., Giuffra V., Duchêne S., Duggan A.T. The paradox of HBV evolution as revealed from a 16thcentury mummy. PLoS Pathog. 2018;14:1–25. doi: 10.1371/journal.ppat.1006750.
- Mühlemann B., Jones T.C., De Barros Damgaard P., Allentoft M.E., Shevnina I., Logvin A. Ancient hepatitis B viruses from the bronze age to the medieval period. Nature. 2018;557:418–423. doi: 10.1038/s41586-018-0097-z.
- Flicker S., Steinberger P., Norderhaug L., Sperr W.R., Majlesi Y., Valent P. Conversion of grass pollen allergen-specific human IgE into a protective IgG(1) antibody. Eur J Immunol. 2002;32:2156–2162. doi: 10.1002/1521-4141(200208)32:8<2156::AID-IMMU2156>;2-A.
- Madritsch C., Flicker S., Scheiblhofer S., Zafred D., Pavkov-Keller T., Thalhamer J. Recombinant monoclonal human immunoglobulin E to investigate the allergenic activity of major grass pollen allergen Phl p 5. Clin Exp Allergy. 2011;41:270–280. doi: 10.1111/j.1365-2222.2010.03666.x.
- Babicki S., Arndt D., Marcu A., Liang Y., Grant J.R., Maciejewski A. Heatmapper: web-enabled heat mapping for all. Nucleic Acids Res. 2016;44:W147–W153. doi: 10.1093/nar/gkw419.
- Velkov S., Ott J., Protzer U., Michler T. The global hepatitis B virus genotype distribution approximated from available genotyping data. Genes. 2018;9:495. doi: 10.3390/genes9100495.
- Herrscher C., Roingeard P., Blanchard E. Hepatitis B virus entry into cells. Cells. 2020;9:1486. doi: 10.3390/cells9061486.
- Pol S., Nalpas B., Driss F., Michel M.-.L., Tiollais P., Denis J. Efficacy and limitations of a specific immunotherapy in chronic hepatitis B. J Hepatol. 2001;34:917–921. doi: 10.1016/S0168-8278(01)00028-9.
- Wang W., Zhou X., Bian Y., Wang S., Chai Q., Guo Z. Dual-targeting nanoparticle vaccine elicits a therapeutic antibody response against chronic hepatitis B. Nat Nanotechnol. 2020;15(5):406–416. doi: 10.1038/s41565-020-0648-y.
- Bourgine M., Dion S., Godon O., Guillen G., Michel M.-.L., Aguilar J.C. Optimization of immune responses induced by therapeutic vaccination with cross-reactive antigens in a humanized hepatitis B surface antigen transgenic mouse model. Virology. 2012;430:10–19. doi: 10.1016/j.virol.2012.04.007.
- Li D., He W., Liu X., Zheng S., Qi Y., Li H. A potent human neutralizing antibody Fc-dependently reduces established HBV infections. Elife. 2017;6:e26738. doi: 10.7554/eLife.26738.
- Blank A., Markert C., Hohmann N., Carls A., Mikus G., Lehr T. First-in-human application of the novel hepatitis B and hepatitis D virus entry inhibitor myrcludex B. J Hepatol. 2016;65:483–489. doi: 10.1016/j.jhep.2016.04.013.
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