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Alphavirus-Based Antigen Preparation

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Vaccine Delivery Technology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2183))

Abstract

Alphavirus-based vectors present an efficient approach for antigen preparation applied for vaccine development. Semliki Forest virus, Sindbis virus, and Venezuelan equine encephalitis virus have been engineered for high-level expression of antigens targeting infectious diseases and tumors. Alphaviruses possess a large application range as vectors can be delivered as naked RNA replicons, recombinant viral particles, and layered DNA plasmids. Immunization studies in animal models have provided protection against challenges with lethal doses of pathogenic infectious agents and tumor cells. So far, a limited number of clinical trials have been conducted for alphavirus vectors in humans.

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References

  1. Strauss J, Strauss E (1994) The alphaviruses; gene expression, replication and evolution. Microbiol Rev 58:491–562

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Liljestrom P, Garoff H (1991) A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Bio/Technology 9:1356–1361

    Article  CAS  Google Scholar 

  3. Xiong C, Levis R, Shen P, Schlesinger S, Rice CM, Huang HV (1989) Sindbis virus: an efficient, broad host range vector for gene expression in animal cells. Science 243:1188–1191

    Article  CAS  PubMed  Google Scholar 

  4. Davies NL, Brown KW, Johnston RE (1989) In vitro synthesis of infectious Venezuelan equine encephalitis virus RNA from a cDNA clone: analysis of a viable deletion mutant. Virologie 171:189–204

    Article  Google Scholar 

  5. Lundstrom K (2014) Alphavirus-based vaccines. Viruses 6:2392–2415

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Schultz-Cherry S, Dybing JK, Davis NL, Williamson C, Suarez DL, Johnston R et al (2000) Influenza virus (A/HK/156/97) hemagglutinin expressed by an alphavirus replicon system protects against lethal infection with Hong Kong-origin H5N1 viruses. Virology 278:55–59

    Article  CAS  PubMed  Google Scholar 

  7. Fleeton MN, Chen M, Berglund P, Rhodes G, Parker SE, Murphy M et al (2001) Self-replicative RNA vaccines elicit protection against influenza A virus, respiratory syncytial virus, and a tickborne encephalitis virus. J Infect Dis 183:1395–1398

    Article  CAS  PubMed  Google Scholar 

  8. Wang M, Jokinen J, Tretyakova I, Pushko P, Luikashevich IS (2018) Alphavirus vector-based replicon particles expressing multivalent cross-protective Lassa virus glycoproteins. Vaccine 36:683–690

    Article  CAS  PubMed  Google Scholar 

  9. Wilson JA, Hart MK (2001) Protection from Ebola virus mediated by cytotoxic T-lymphocytes specific for the viral nucleoprotein. J Virol 75:2660–2664

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Pushko P, Bray M, Ludwig GV, Parker M, Schmaljohn A, Sanchez A et al (2000) Recombinant RNA replicons derived from attenuated Venezuelan equine encephalitis virus protect guinea pigs and mice from Ebola hemorrhagic fever virus. Vaccine 19:142–153

    Article  CAS  PubMed  Google Scholar 

  11. Herbert AS, Kuehne AI, Barth JF, Ortiz RA, Nichols DK, Zak SE et al (2013) Venezuelan equine encephalitis virus replicon particle vaccine protects nonhuman primates from intramuscular and aerosol challenge with Ebolavirus. J Virol 87:4952–4964

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Vogel AB, Lambert L, Kinnear E, Busse D, Erbar S, Reuter KC et al (2018) Self-amplifying RNA vaccines give equivalent protection against influenza to mRNA vaccines but at much lower doses. Mol Ther 26:446–455

    Article  CAS  PubMed  Google Scholar 

  13. Hariharan MJ, Driver DA, Townsend K, Brumm D, Polo JM, Belli BA et al (1998) DNA immunization against herpes simplex virus: enhanced efficacy using a Sindbis virus-based vector. J Virol 72:950–958

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Kirman JR, Turon T, Su H, Li A, Kraus C, Polo JM et al (2003) Enhanced immunogenicity to Mycobacterium tuberculosis by vaccination with an alphavirus plasmid replicon expressing antigen 85A. Infect Immun 71:575–579

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Reddy JR, Kwang J, Varthakavi V, Lechtenberg KF, Minocha HC (1999) Semliki Forest virus vector carrying the bovine viral diarrhea virus NS3 (p80) cDNA induced immune responses in mice and expressed BVDV protein in mammalian cells. Comp Immunol Microbiol Infect Dis 22:231–246

    Article  CAS  PubMed  Google Scholar 

  16. Knudsen ML, Ljungberg K, Tatoud R, Weber J, Esteban M, Liljestrom P (2015) Alphavirus replicon DNA expressing HIV antigens is an excellent prime for boosting with recombinant Ankara (MVA) or with HIV gp140 protein antigen. PLoS One 10:e0117042

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Roche FP, Sheahan BJ, O’Mara SM, Atkins GJ (2010) Semliki Forest virus-mediated gene therapy of the RG2 rat glioma. Neuropathol Appl Neurobiol 36:648–660

    Article  CAS  PubMed  Google Scholar 

  18. Ylosmaki E, Martikainen M, Hinkkanen A, Saksela K (2013) Attenuation of Semliki Forest virus neurovirulence by microRNA-mediated detargeting. J Virol 87:335–344

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lin Y, Zhang H, Liang J, Li K, Zhu W, Fu L, Wang F, Zheng X, Shi H, Wu S et al (2014) Identification and characterization of alphavirus M1 as a selective oncolytic virus targeting ZAP-defective human cancers. Proc Natl Acad Sci U S A 111:E4504–E4512

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ying H, Zaks TZ, Wang R-F, Irvine KR, Kammula US, Marincola FM et al (1999) Cancer therapy using a self-replicating RNA vaccine. Nat Med 5:823–827

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Van de Wall S, Ljungberg K, Ip PP, Boerma A, Knudsen ML, Nijman HW et al (2018) Potent therapeutic efficacy of an alphavirus replicon DNA vaccine expressing human papilloma virus E6 and E7 antigens. Oncoimmunology 7:e1487913

    Article  PubMed  PubMed Central  Google Scholar 

  22. Leslie MC, Zhao YJ, Lachman LB, Hwu P, Wu GJ, Bar-Eli M (2007) Immunization against MUC18/MCAM, a novel antigen that drives melanoma invasion and metastasis. Gene Ther 14:316–323

    Article  CAS  PubMed  Google Scholar 

  23. Bernstein DI, Reap EA, Katen K, Watson A, Smith K, Norberg P et al (2010) Randomized, double-blind, Phase I trial on an alphavirus replicon vaccine for cytomegalovirus in CMV negative volunteers. Vaccine 28:484–493

    Article  CAS  Google Scholar 

  24. Wecker M, Gilbert P, Russell N, Hural J, Allen M, Pensiero M et al (2012) Phase I safety and immunogenicity evaluations of an alphavirus replicon HIV-1 subtype C gag vaccine in healthy HIV-1-uninfected adults. Clin. Vaccine Immunol 19:1651–1660

    Article  CAS  Google Scholar 

  25. Morse MA, Hobelka AC, Osada T, Berglund P, Hubby B, Negri S et al (2010) An alphavirus vector overcomes the presence of neutralizing antibodies and elevated numbers of Tregs to induce immune responses in humans with advanced cancer. J Clin Investig 120:3234–3241

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Slovin SF, Kehoe M, Durso R, Fernandez C, Olson W, Gao JP et al (2013) A phase I dose escalation trial of vaccine replicon particles (VRP) expressing prostate-specific membrane antigen (PSMA) in subjects with prostate cancer. Vaccine 31:943–949

    Article  CAS  PubMed  Google Scholar 

  27. Ehrengruber MU, Lundstrom K (2016) Recombinant alphavirus-mediated expression of ion channels and receptors in the brain. In: Luján R, Ciruela F (eds) Receptor and ion channel detection in the brain: methods and protocols, Neuromethods, vol 110. Springer Science + Business Media, New York, NY. https://doi.org/10.1007/978-1-4939-3064-7_7

    Chapter  Google Scholar 

  28. Schweitzer C, Kratzeisen C, Adam G, Lundstrom K, Malherbe P, Ohresser S et al (2000) Characterization of [3H] LY-354740 binding of rat mGlu2 and mGlu3 receptors expressed in CHO cells using Semliki Forest virus vectors. Neuropharmacology 39:1700–1706

    Article  CAS  PubMed  Google Scholar 

  29. Berglund P, Sjöberg M, Garoff H, Atkins GJ, Sheahan BJ, Liljestrom P (1993) Semliki Forest virus expression system: production of conditionally infectious recombinant particles. Bio/Technology 11:916–920

    CAS  Google Scholar 

  30. Liljestrom P, Lusa S, Huylebroeck D, Garoff H (1991) In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular weight membrane protein modulates virus release. J Virol 65:4107–4113

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Vähä-Koskela MJ, Tuittila MT, Nygardas PT, Nyman JK, Ehrengruber MU, Renggli M (2003) A novel neurotropic expression vector based on the avirulent A7(74) strain of Semliki Forest virus. J Neurovirol 9:1–15

    Article  PubMed  CAS  Google Scholar 

  32. DiCiommo DP, Bremner R (1998) Rapid, high-level protein production using DNA-based Semliki Forest virus vectors. J Biol Chem 273:18060–18066

    Article  CAS  PubMed  Google Scholar 

  33. DiCiommo DP, Duckett A, Burcescu I, Bremner R, Gallie BR (2004) Retinoblastoma protein purification and transduction of retina and retinoblastoma cells using improved alphavirus vectors. Invest. Ophthalmol Vis Sci 45:3320–3329

    Article  Google Scholar 

  34. Ulper L, Sarand I, Rausalu K, Merits A (2008) Construction, properties, and potential application of infectious plasmids containing Semliki Forest virus full-length cDNA with a n inserted intron. J Virol Methods 148:265–270

    Article  CAS  PubMed  Google Scholar 

  35. Puglia AL, Rezende AG, Jorge SA, Wagner R, Pereira CA, Astray RM (2013) Quantitative RT-PCR for titration of replication-defective recombinant Semliki Forest virus. J Virol Methods 193:647–652

    Article  CAS  PubMed  Google Scholar 

  36. Juare D, Long KC, Aguilar P, Kochel TJ, Halsey JS (2013) Assessment of plaque assay methods for alphaviruses. J Virol Methods 187:185–189

    Article  CAS  Google Scholar 

  37. Avogadri F, Merghoub T, Maughan MF, Hirschhorn-Cymerman D, Morris J, Ritter E et al (2010) Alphavirus replicon particles expressing TRP-2 provide potent therapeutic effect on melanoma through activation of humoral and cellular immunity. PLoS One 5:e12670

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  38. Leitner WW, Hwang NL, de Veer MJ, Zhou A et al (2003) Alphavirus-based DNA vaccine breaks immunological tolerance by activating innate antiviral pathways. Nat Med 5:33–39

    Article  CAS  Google Scholar 

  39. Colombage G, Hall R, Pavy M, Lobigs M (1998) DNA-based and alphavirus-vectored immunization with PrM and E proteins elicits long-lived and protective immunity against the flavivirus, Murray Valley encephalitis virus. Virology 250:151–163

    Article  CAS  PubMed  Google Scholar 

  40. Lundstrom K, Abenavoli A, Malgaroli A, Ehrengruber MU (2003) Novel Semliki Forest virus vectors with reduced toxicity and temperature-sensitivity for long-term enhancement of transgene expression. Mol Ther 7:202–209

    Article  CAS  PubMed  Google Scholar 

  41. Agapov EV, Frolov I, Lindenbach BD, Pragal PM, Schlesinger S, Rice CM (1998) Noncythopathic Sindbis virus RNA vectors for heterologous gene expression. Proc Natl Acad Sci U S A 95:12989–12994

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Sjöberg EM, Suomalainen M, Garoff H (1994) A significantly improved Semliki Forest virus expression system based on translation enhancer segments from the viral capsid gene. Bio/Technology 12:1127–1131

    Article  Google Scholar 

  43. Geall AJ, Verma A, Otten GR, Shaw CA, Hekele A, Banerjee K et al (2012) Nonviral delivery of self-amplifying RNA vaccines. Proc Natl Acad Sci U S A 109:14604–14609

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Voytas D, Ke N (2001) Detection and quantitation of radiolabeled proteins and DNA in gels and blot. Curr Protoc Mol Biol Appendix 3:3A

    Google Scholar 

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Author information

Authors and Affiliations

  1. PanTherapeutics, Lutry, Switzerland

    Kenneth Lundstrom

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  1. Kenneth Lundstrom
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Editor information

Editors and Affiliations

  1. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, NY, USA

    Blaine A. Pfeifer

  2. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, NY, USA

    Andrew Hill

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Lundstrom, K. (2021). Alphavirus-Based Antigen Preparation. In: Pfeifer, B.A., Hill, A. (eds) Vaccine Delivery Technology. Methods in Molecular Biology, vol 2183. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0795-4_6

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  • DOI: https://doi.org/10.1007/978-1-0716-0795-4_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0794-7

  • Online ISBN: 978-1-0716-0795-4

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