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Open Access 01-12-2022 | Rabies | Research

An mRNA-based rabies vaccine induces strong protective immune responses in mice and dogs

Authors: Jianglong Li, Qi Liu, Jun Liu, Xiaohong Wu, Yixin Lei, Shuang Li, Danhua Zhao, Zhi Li, Liping Luo, Sophia Peng, Yingrao Ou, Hong Yang, Jing Jin, Yuhua Li, Yucai Peng

Published in: Virology Journal | Issue 1/2022

Abstract

Rabies is a lethal zoonotic disease that is mainly caused by the rabies virus (RABV). Although effective vaccines have long existed, current vaccines take both time and cost to produce. Messenger RNA (mRNA) technology is an emergent vaccine platform that supports rapid vaccine development on a large scale. Here, an optimized mRNA vaccine construct (LVRNA001) expressing rabies virus glycoprotein (RABV-G) was developed in vitro and then evaluated in vivo for its immunogenicity and protective capacity in mice and dogs. LVRNA001 induced neutralizing antibody production and a strong Th1 cellular immune response in mice. In both mice and dogs, LVRNA001 provided protection against challenge with 50-fold lethal dose 50 (LD₅₀) of RABV. With regards to protective efficiency, an extended dosing interval (14 days) induced greater antibody production than 3- or 7-day intervals in mice. Finally, post-exposure immunization against RABV was performed to evaluate the survival rates of dogs receiving two 25 μg doses of LVRNA001 vs. five doses of inactivated vaccine over the course of three months. Survival rate in the LVRNA001 group was 100%, whereas survival rate in the inactivated vaccine control group was only 33.33%. In conclusion, these results demonstrated that LVRNA001 induced strong protective immune responses in mice and dogs, which provides a new and promising prophylactic strategy for rabies.

Bourhy H, Sureau P, Tordo N. From rabies to rabies-related viruses. Vet Microbiol. 1990;23:115–28. https://doi.org/10.1016/0378-1135(90)90141-h.CrossRefPubMed

Ray NB, Ewalt LC, Lodmell DL. Rabies virus replication in primary murine bone marrow macrophages and in human and murine macrophage-like cell lines: implications for viral persistence. J Virol. 1995;69:764–72. https://doi.org/10.1128/JVI.69.2.764-772.1995.CrossRefPubMedPubMedCentral

Ugolini G, Hemachudha T. Rabies: changing prophylaxis and new insights in pathophysiology. Curr Opin Infect Dis. 2018;31:93–101. https://doi.org/10.1097/QCO.0000000000000420.CrossRefPubMed

Hampson K, Coudeville L, Lembo T, Sambo M, Kieffer A, Attlan M, Barrat J, Blanton JD, Briggs DJ, Cleaveland S, et al. Estimating the global burden of endemic canine rabies. PLoS Negl Trop Dis. 2015;9: e0003709. https://doi.org/10.1371/journal.pntd.0003709.CrossRefPubMedPubMedCentral

Davis BM, Rall GF, Schnell MJ. Everything you always wanted to know about rabies virus (But Were Afraid to Ask). Ann Rev Virol. 2015;2:451–71. https://doi.org/10.1146/annurev-virology-100114-055157.CrossRef

Fisher CR, Streicker DG, Schnell MJ. The spread and evolution of rabies virus: conquering new frontiers. Nat Rev Microbiol. 2018;16:241–55. https://doi.org/10.1038/nrmicro.2018.11.CrossRefPubMedPubMedCentral

Tordo N, Poch O, Ermine A, Keith G, Rougeon F. Walking along the rabies genome: is the large G-L intergenic region a remnant gene? Proc Natl Acad Sci USA. 1986;83:3914–8. https://doi.org/10.1073/pnas.83.11.3914.CrossRefPubMedPubMedCentral

Wunner WH, Reagan KJ, Koprowski H. Characterization of saturable binding sites for rabies virus. J Virol. 1984;50:691–7. https://doi.org/10.1128/JVI.50.3.691-697.1984.CrossRefPubMedPubMedCentral

Thoulouze MI, Lafage M, Schachner M, Hartmann U, Cremer H, Lafon M. The neural cell adhesion molecule is a receptor for rabies virus. J Virol. 1998;72:7181–90. https://doi.org/10.1128/JVI.72.9.7181-7190.1998.CrossRefPubMedPubMedCentral

10.

Tuffereau C, Benejean J, Blondel D, Kieffer B, Flamand A. Low-affinity nerve-growth factor receptor (P75NTR) can serve as a receptor for rabies virus. EMBO J. 1998;17:7250–9. https://doi.org/10.1093/emboj/17.24.7250.CrossRefPubMedPubMedCentral

11.

Gaudin Y, Tuffereau C, Durrer P, Brunner J, Flamand A, Ruigrok R. Rabies virus-induced membrane fusion. Mol Membr Biol. 1999;16:21–31. https://doi.org/10.1080/096876899294724.CrossRefPubMed

12.

Albertini AA, Baquero E, Ferlin A, Gaudin Y. Molecular and cellular aspects of rhabdovirus entry. Viruses. 2012;4:117–39. https://doi.org/10.3390/v4010117.CrossRefPubMedPubMedCentral

13.

Etessami R, Conzelmann KK, Fadai-Ghotbi B, Natelson B, Tsiang H, Ceccaldi PE. Spread and pathogenic characteristics of a G-deficient rabies virus recombinant: an in vitro and in vivo study. J Gen Virol. 2000;81:2147–53. https://doi.org/10.1099/0022-1317-81-9-2147.CrossRefPubMed

14.

Kucera P, Dolivo M, Coulon P, Flamand A. Pathways of the early propagation of virulent and avirulent rabies strains from the eye to the brain. J Virol. 1985;55:158–62. https://doi.org/10.1128/JVI.55.1.158-162.1985.CrossRefPubMedPubMedCentral

15.

Flamand A, Raux H, Gaudin Y, Ruigrok RW. Mechanisms of rabies virus neutralization. Virology. 1993;194:302–13. https://doi.org/10.1006/viro.1993.1261.CrossRefPubMed

16.

Prehaud C, Coulon P, LaFay F, Thiers C, Flamand A. Antigenic site II of the rabies virus glycoprotein: structure and role in viral virulence. J Virol. 1988;62:1–7. https://doi.org/10.1128/JVI.62.1.1-7.1988.CrossRefPubMedPubMedCentral

17.

Seif I, Coulon P, Rollin PE, Flamand A. Rabies virulence: effect on pathogenicity and sequence characterization of rabies virus mutations affecting antigenic site III of the glycoprotein. J Virol. 1985;53:926–34. https://doi.org/10.1128/JVI.53.3.926-934.1985.CrossRefPubMedPubMedCentral

18.

Zhu S, Guo C. Rabies control and treatment: from prophylaxis to strategies with curative potential. Viruses. 2016. https://doi.org/10.3390/v8110279.CrossRefPubMedPubMedCentral

19.

Chen C, Zhang C, Li R, Wang Z, Yuan Y, Li H, Fu Z, Zhou M, Zhao L. Monophosphoryl-Lipid A (MPLA) is an efficacious adjuvant for inactivated rabies vaccines. Viruses. 2019. https://doi.org/10.3390/v11121118.CrossRefPubMedPubMedCentral

20.

Rupprecht CE, Blass L, Smith K, Orciari LA, Niezgoda M, Whitfield SG, Gibbons RV, Guerra M, Hanlon CA. Human infection due to recombinant vaccinia-rabies glycoprotein virus. N Engl J Med. 2001;345:582–6. https://doi.org/10.1056/NEJMoa010560.CrossRefPubMed

21.

Rupprecht CE, Hanlon CA, Blanton J, Manangan J, Morrill P, Murphy S, Niezgoda M, Orciari LA, Schumacher CL, Dietzschold B. Oral vaccination of dogs with recombinant rabies virus vaccines. Virus Res. 2005;111:101–5. https://doi.org/10.1016/j.virusres.2005.03.017.CrossRefPubMed

22.

Rupprecht CE, Charlton KM, Artois M, Casey GA, Webster WA, Campbell JB, Lawson KF, Schneider LG. Ineffectiveness and comparative pathogenicity of attenuated rabies virus vaccines for the striped skunk (Mephitis mephitis). J Wildl Dis. 1990;26:99–102. https://doi.org/10.7589/0090-3558-26.1.99.CrossRefPubMed

23.

Jackson NAC, Kester KE, Casimiro D, Gurunathan S, DeRosa F. The promise of mRNA vaccines: a biotech and industrial perspective. NPJ Vaccines. 2020;5:11. https://doi.org/10.1038/s41541-020-0159-8.CrossRefPubMedPubMedCentral

24.

Mascola JR, Fauci AS. Novel vaccine technologies for the 21st century. Nat Rev Immunol. 2020;20:87–8. https://doi.org/10.1038/s41577-019-0243-3.CrossRefPubMed

25.

Maruggi G, Zhang C, Li J, Ulmer JB, Yu D. mRNA as a transformative technology for vaccine development to control infectious diseases. Mol Ther J Am Soc Gene Ther. 2019;27:757–72. https://doi.org/10.1016/j.ymthe.2019.01.020.CrossRef

26.

Amanpour S. The rapid development and early success of Covid 19 vaccines have raised hopes for accelerating the cancer treatment mechanism. Arch Razi Inst. 2021;76:1–6. https://doi.org/10.22092/ari.2021.353761.1612.CrossRefPubMedPubMedCentral

27.

Vasireddy D, Atluri P, Malayala SV, Vanaparthy R, Mohan G. Review of COVID-19 vaccines approved in the united states of america for emergency use. J Clin Med Res. 2021;13:204–13. https://doi.org/10.14740/jocmr4490.CrossRefPubMedPubMedCentral

28.

Alameh MG, Weissman D, Pardi N. Messenger RNA-based vaccines against infectious diseases. Curr Top Microbiol Immunol. 2020. https://doi.org/10.1007/82_2020_202.CrossRefPubMed

29.

Hoerr I, Obst R, Rammensee HG, Jung G. In vivo application of RNA leads to induction of specific cytotoxic T lymphocytes and antibodies. Eur J Immunol. 2000;30:1–7. https://doi.org/10.1002/1521-4141(200001)30:1%3c1::AID-IMMU1%3e3.0.CO;2-#.CrossRefPubMed

30.

Heine A, Juranek S, Brossart P. Clinical and immunological effects of mRNA vaccines in malignant diseases. Mol Cancer. 2021;20:52. https://doi.org/10.1186/s12943-021-01339-1.CrossRefPubMedPubMedCentral

31.

Wang Y, Zhang Z, Luo J, Han X, Wei Y, Wei X. mRNA vaccine: a potential therapeutic strategy. Mol Cancer. 2021;20:33. https://doi.org/10.1186/s12943-021-01311-z.CrossRefPubMedPubMedCentral

32.

Pardi N, Hogan MJ, Porter FW, Weissman D. mRNA vaccines - a new era in vaccinology. Nat Rev Drug Discov. 2018;17:261–79. https://doi.org/10.1038/nrd.2017.243.CrossRefPubMedPubMedCentral

33.

Alberer M, Gnad-Vogt U, Hong HS, Mehr KT, Backert L, Finak G, Gottardo R, Bica MA, Garofano A, Koch SD, et al. Safety and immunogenicity of a mRNA rabies vaccine in healthy adults: an open-label, non-randomised, prospective, first-in-human phase 1 clinical trial. Lancet. 2017;390:1511–20. https://doi.org/10.1016/S0140-6736(17)31665-3.CrossRefPubMed

34.

Aldrich C, Leroux-Roels I, Huang KB, Bica MA, Loeliger E, Schoenborn-Kellenberger O, Walz L, Leroux-Roels G, von Sonnenburg F, Oostvogels L. Proof-of-concept of a low-dose unmodified mRNA-based rabies vaccine formulated with lipid nanoparticles in human volunteers: a phase 1 trial. Vaccine. 2021;39:1310–8. https://doi.org/10.1016/j.vaccine.2020.12.070.CrossRefPubMedPubMedCentral

35.

Miao FM, Zhang SF, Wang SC, Liu Y, Zhang F, Hu RL. Comparison of immune responses to attenuated rabies virus and street virus in mouse brain. Adv Virol. 2017;162:247–57. https://doi.org/10.1007/s00705-016-3081-7.CrossRef

36.

Zhao J, Wang S, Zhang S, Liu Y, Zhang J, Zhang F, Mi L, Hu R. Molecular characterization of a rabies virus isolate from a rabid dog in Hanzhong District, Shaanxi Province, China. Arch Virol. 2014;159:1481–6. https://doi.org/10.1007/s00705-013-1941-y.CrossRefPubMed

37.

Du J, Zhang Q, Tang Q, Li H, Tao X, Morimoto K, Nadin-Davis SA, Liang G. Characterization of human rabies virus vaccine strain in China. Virus Res. 2008;135:260–6. https://doi.org/10.1016/j.virusres.2008.04.002.CrossRefPubMed

38.

Ren L. Molecular characterization of a Chinese variant of the Flury-LEP strain. Virol J. 2010;7:80. https://doi.org/10.1186/1743-422X-7-80.CrossRefPubMedPubMedCentral

39.

Jia L, Liu YP, Tian LF, Xiong C, Xu X, Qu H, Xiong W, Zhou D, Wang F, Liu Z, et al. Potent neutralizing RBD-specific antibody cocktail against SARS-CoV-2 and its mutant. MedComm. 2021. https://doi.org/10.1002/mco2.79.CrossRefPubMedPubMedCentral

40.

He C, Yang J, He X, Hong W, Lei H, Chen Z, Shen G, Yang L, Li J, Wang Z, et al. A bivalent recombinant vaccine targeting the S1 protein induces neutralizing antibodies against both SARS-CoV-2 variants and wild-type of the virus. MedComm. 2021. https://doi.org/10.1002/mco2.72.CrossRefPubMedPubMedCentral

41.

Cox JH, Dietzschold B, Schneider LG. Rabies virus glycoprotein. II. Biological and serological characterization. Infect Immun. 1977;16:754–9. https://doi.org/10.1128/iai.16.3.754-759.1977.CrossRefPubMedPubMedCentral

42.

Dietzschold B, Wiktor TJ, Macfarlan R, Varrichio A. Antigenic structure of rabies virus glycoprotein: ordering and immunological characterization of the large CNBr cleavage fragments. J Virol. 1982;44:595–602. https://doi.org/10.1128/JVI.44.2.595-602.1982.CrossRefPubMedPubMedCentral

43.

Lutz J, Lazzaro S, Habbeddine M, Schmidt KE, Baumhof P, Mui BL, Tam YK, Madden TD, Hope MJ, Heidenreich R, et al. Unmodified mRNA in LNPs constitutes a competitive technology for prophylactic vaccines. NPJ vaccines. 2017;2:29. https://doi.org/10.1038/s41541-017-0032-6.CrossRefPubMedPubMedCentral

44.

Petsch B, Schnee M, Vogel AB, Lange E, Hoffmann B, Voss D, Schlake T, Thess A, Kallen KJ, Stitz L, et al. Protective efficacy of in vitro synthesized, specific mRNA vaccines against influenza A virus infection. Nat Biotechnol. 2012;30:1210–6. https://doi.org/10.1038/nbt.2436.CrossRefPubMed

45.

Kieny MP, Lathe R, Drillien R, Spehner D, Skory S, Schmitt D, Wiktor T, Koprowski H, Lecocq JP. Expression of rabies virus glycoprotein from a recombinant vaccinia virus. Nature. 1984;312:163–6. https://doi.org/10.1038/312163a0.CrossRefPubMed

46.

Ge J, Wang X, Tao L, Wen Z, Feng N, Yang S, Xia X, Yang C, Chen H, Bu Z. Newcastle disease virus-vectored rabies vaccine is safe, highly immunogenic, and provides long-lasting protection in dogs and cats. J Virol. 2011;85:8241–52. https://doi.org/10.1128/JVI.00519-11.CrossRefPubMedPubMedCentral

47.

Ross J, Sullivan TD. Half-lives of beta and gamma globin messenger RNAs and of protein synthetic capacity in cultured human reticulocytes. Blood. 1985;66:1149–54.CrossRefPubMed

48.

Holtkamp S, Kreiter S, Selmi A, Simon P, Koslowski M, Huber C, Tureci O, Sahin U. Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. Blood. 2006;108:4009–17. https://doi.org/10.1182/blood-2006-04-015024.CrossRefPubMed

49.

Gallie DR. The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev. 1991;5:2108–16. https://doi.org/10.1101/gad.5.11.2108.CrossRefPubMed

50.

Thess A, Grund S, Mui BL, Hope MJ, Baumhof P, Fotin-Mleczek M, Schlake T. Sequence-engineered mRNA without chemical nucleoside modifications enables an effective protein therapy in large animals. Mol Ther J Am Soc Gene Ther. 2015;23:1456–64. https://doi.org/10.1038/mt.2015.103.CrossRef

51.

Kudla G, Lipinski L, Caffin F, Helwak A, Zylicz M. High guanine and cytosine content increases mRNA levels in mammalian cells. PLoS Biol. 2006;4: e180. https://doi.org/10.1371/journal.pbio.0040180.CrossRefPubMedPubMedCentral

52.

Cher DJ, Mosmann TR. Two types of murine helper T cell clone. II. Delayed-type hypersensitivity is mediated by TH1 clones. J Immunol. 1987;138:3688–94.PubMed

53.

Coffman RL, Seymour BW, Lebman DA, Hiraki DD, Christiansen JA, Shrader B, Cherwinski HM, Savelkoul HF, Finkelman FD, Bond MW, et al. The role of helper T cell products in mouse B cell differentiation and isotype regulation. Immunol Rev. 1988;102:5–28. https://doi.org/10.1111/j.1600-065x.1988.tb00739.x.CrossRefPubMed

54.

Mosmann TR, Cherwinski H, Bond MW, Giedlin MA, Coffman RL. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol. 1986;136:2348–57.PubMed

55.

Lappin MB, Campbell JD. The Th1-Th2 classification of cellular immune responses: concepts, current thinking and applications in haematological malignancy. Blood Rev. 2000;14:228–39. https://doi.org/10.1054/blre.2000.0136.CrossRefPubMed

56.

Hooper DC, Roy A, Barkhouse DA, Li J, Kean RB. Rabies virus clearance from the central nervous system. Adv Virus Res. 2011;79:55–71. https://doi.org/10.1016/B978-0-12-387040-7.00004-4.CrossRefPubMed

57.

Lebrun A, Portocarrero C, Kean RB, Barkhouse DA, Faber M, Hooper DC. T-bet is required for the rapid clearance of attenuated rabies virus from central nervous system tissue. J Immunol. 2015;195:4358–68. https://doi.org/10.4049/jimmunol.1501274.CrossRefPubMed

58.

Payne RP, Longet S, Austin JA, Skelly DT, Dejnirattisai W, Adele S, Meardon N, Faustini S, Al-Taei S, Moore SC, et al. Immunogenicity of standard and extended dosing intervals of BNT162b2 mRNA vaccine. Cell. 2021;184:5699-5714 e5611. https://doi.org/10.1016/j.cell.2021.10.011.CrossRefPubMedPubMedCentral

Title: An mRNA-based rabies vaccine induces strong protective immune responses in mice and dogs
Authors: Jianglong Li
Qi Liu
Jun Liu
Xiaohong Wu
Yixin Lei
Shuang Li
Danhua Zhao
Zhi Li
Liping Luo
Sophia Peng
Yingrao Ou
Hong Yang
Jing Jin
Yuhua Li
Yucai Peng
Publication date: 01-12-2022
Publisher: BioMed Central
Keyword: Rabies
Published in: Virology Journal / Issue 1/2022
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/s12985-022-01919-7

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