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Alphavirus Replicon Vectors for Prophylactic Applications and Cancer Intervention

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Novel Technologies for Vaccine Development

Abstract

Alphavirus replicons represent self-replicating RNA molecules resembling alphaviral genomic RNA, except replicons encode antigen(s) of interest in place of an alphaviral structural polyprotein. Because viral structural genes are missing, replicon RNA cannot initiate replication of an alphavirus. However, due to the presence of intrinsic RNA-dependent RNA polymerase activity, replicons are capable of self-amplification in vitro and in vivo resulting in high levels of expression of antigen of interest. For vaccination or therapeutic purposes, replicons can be delivered in vivo by replicon particles. The latter represent viruslike particle vectors (VLPVs) that encapsidate replicon RNA and deliver it into target cells for antigen expression. The viruslike nature and self-replicating RNA features ensure efficient priming of innate immunity and adjuvant effect, while high-level expression provides antigen for induction of cell-mediated and humoral immune responses. Replicon vectors have been developed from several alphaviruses including Venezuelan equine encephalitis virus (VEEV), Semliki Forest virus (SFV), and Sindbis virus (SINV). Applications of replicon particles included prophylactic and therapeutic vaccines for infectious diseases and cancer, as well as adjuvants for enhancement of immune responses. In several preclinical models including nonhuman primates, alphavirus replicons have shown exceptional promise as safe and effective vaccines and adjuvants. Experimental replicon vaccines included vaccines against influenza, Ebola, Marburg, and Lassa viruses. Bivalent vaccines protecting from both Ebola and Lassa viruses have been also described. Protective effects have been reported for cancer indications after therapeutic vaccination with replicon vaccines expressing tumor-associated antigens. Clinical trials involving alphavirus replicons are underway. In this review, an attempt is made to summarize the state of the art of the alphavirus replicon-based technology for prophylactic and therapeutic applications. The advantages and challenges of the replicon technologies are presented, and the future of this promising platform is discussed.

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References

  • Berglund P, Quesada-Rolander M, Putkonen P, Biberfeld G, Thorstensson R, Liljestrom P (1997) Outcome of immunization of cynomolgus monkeys with recombinant Semliki Forest virus encoding human immunodeficiency virus type 1 envelope protein and challenge with a high dose of SHIV-4 virus. AIDS Res Hum Retrovir 13(17):1487–1495

    Article  CAS  PubMed  Google Scholar 

  • Berglund P, Smerdou C, Fleeton MN, Tubulekas I, Liljestrom P (1998) Enhancing immune responses using suicidal DNA vaccines. Nat Biotechnol 16(6):562–565

    Article  CAS  PubMed  Google Scholar 

  • Berglund P, Fleeton MN, Smerdou C, Liljestrom P (1999) Immunization with recombinant Semliki Forest virus induces protection against influenza challenge in mice. Vaccine 17(5):497–507

    Article  CAS  PubMed  Google Scholar 

  • Bernstein DI, Reap EA, Katen K, Watson A, Smith K, Norberg P et al (2009) Randomized, double-blind, Phase 1 trial of an alphavirus replicon vaccine for cytomegalovirus in CMV seronegative adult volunteers. Vaccine 28(2):484–493

    Article  CAS  PubMed  Google Scholar 

  • Bosworth B, Erdman MM, Stine DL, Harris I, Irwin C, Jens M et al (2010) Replicon particle vaccine protects swine against influenza. Comp Immunol Microbiol Infect Dis 33(6):e99–e103

    Article  CAS  PubMed  Google Scholar 

  • Cao S, Zhang W (2013) Characterization of an early-stage fusion intermediate of Sindbis virus using cryoelectron microscopy. Proc Natl Acad Sci U S A 110(33):13362–13367

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Carrion R Jr, Bredenbeek P, Jiang X, Tretyakova I, Pushko P, Lukashevich IS (2012) Vaccine platforms to control arenaviral hemorrhagic fevers. J Vaccines Vaccin 20:3(7)

    Google Scholar 

  • Carroll TD, Matzinger SR, Barro M, Fritts L, McChesney MB, Miller CJ et al (2011) Alphavirus replicon-based adjuvants enhance the immunogenicity and effectiveness of Fluzone (R) in rhesus macaques. Vaccine 29(5):931–940

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Chang CF, Wang M, Ou WC, Chen PL, Shen CH, Lin PY et al (2011) Human JC virus-like particles as a gene delivery vector. Expert Opin Biol Ther 11(9):1169–1175

    Article  CAS  PubMed  Google Scholar 

  • Chen LH, Wilson ME (2012) Dengue and chikungunya in travelers: recent updates. Curr Opin Infect Dis 25(5):523–529

    Article  CAS  PubMed  Google Scholar 

  • Dahiya SS, Saini M, Kumar P, Gupta PK (2011) An oral Sindbis virus replicon-based DNA vaccine containing VP2 gene of canine parvovirus delivered by Escherichia coli elicits immune responses in dogs. Acta Virol 55(4):289–294

    Article  CAS  PubMed  Google Scholar 

  • Davis NL, Willis LV, Smith JF, Johnston RE (1989) In vitro synthesis of infectious venezuelan equine encephalitis virus RNA from a cDNA clone: analysis of a viable deletion mutant. Virology 171(1):189–204

    Article  CAS  PubMed  Google Scholar 

  • Davis NL, Grieder FB, Smith JF, Greenwald GF, Valenski ML, Sellon DC et al (1994) A molecular genetic approach to the study of Venezuelan equine encephalitis virus pathogenesis. Arch Virol Suppl 9:99–109

    CAS  PubMed  Google Scholar 

  • Davis NL, Brown KW, Johnston RE (1996) A viral vaccine vector that expresses foreign genes in lymph nodes and protects against mucosal challenge. J Virol 70(6):3781–3787

    CAS  PubMed Central  PubMed  Google Scholar 

  • Dubensky TW Jr, Driver DA, Polo JM, Belli BA, Latham EM, Ibanez CE et al (1996) Sindbis virus DNA-based expression vectors: utility for in vitro and in vivo gene transfer. J Virol 70(1):508–519

    CAS  PubMed Central  PubMed  Google Scholar 

  • Eriksson M, Andreasson K, Weidmann J, Lundberg K, Tegerstedt K, Dalianis T et al (2011) Murine polyomavirus virus-like particles carrying full-length human PSA protect BALB/c mice from outgrowth of a PSA expressing tumor. PLoS One 6(8):e23828

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Frolov I, Hoffman TA, Pragai BM, Dryga SA, Huang HV, Schlesinger S et al (1996) Alphavirus-based expression vectors: strategies and applications. Proc Natl Acad Sci U S A 93(21):11371–11377

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Frolov I, Frolova E, Schlesinger S (1997) Sindbis virus replicons and Sindbis virus: assembly of chimeras and of particles deficient in virus RNA. J Virol 71(4):2819–2829

    CAS  PubMed Central  PubMed  Google Scholar 

  • Geisbert TW, Pushko P, Anderson K, Smith J, Davis KJ, Jahrling PB (2002) Evaluation in nonhuman primates of vaccines against Ebola virus. Emerg Infect Dis 8(5):503–507

    Article  PubMed Central  PubMed  Google Scholar 

  • Gibbons DL, Vaney MC, Roussel A, Vigouroux A, Reilly B, Lepault J et al (2004) Conformational change and protein-protein interactions of the fusion protein of Semliki Forest virus. Nature 427(6972):320–325

    Article  CAS  PubMed  Google Scholar 

  • Hahn CS, Hahn YS, Braciale TJ, Rice CM (1992) Infectious Sindbis virus transient expression vectors for studying antigen processing and presentation. Proc Natl Acad Sci U S A 89(7):2679–2683

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • 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(9):4952–4964

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hevey M, Negley D, Pushko P, Smith J, Schmaljohn A (1998) Marburg virus vaccines based upon alphavirus replicons protect guinea pigs and nonhuman primates. Virology 251(1):28–37

    Article  CAS  PubMed  Google Scholar 

  • Johanning FW, Conry RM, LoBuglio AF, Wright M, Sumerel LA, Pike MJ et al (1995) A Sindbis virus mRNA polynucleotide vector achieves prolonged and high level heterologous gene expression in vivo. Nucleic Acids Res 23(9):1495–1501

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Johansson DX, Ljungberg K, Kakoulidou M, Liljestrom P (2012) Intradermal electroporation of naked replicon RNA elicits strong immune responses. PLoS One 7(1):e29732

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kamrud KI, Alterson KD, Andrews C, Copp LO, Lewis WC, Hubby B et al (2008) Analysis of Venezuelan equine encephalitis replicon particles packaged in different coats. PLoS One 3(7):e2709

    Article  PubMed Central  PubMed  Google Scholar 

  • Kielian M (2010) Structural biology: an alphavirus puzzle solved. Nature 468(7324):645–646

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Knudsen ML, Mbewe-Mvula A, Rosario M, Johansson DX, Kakoulidou M, Bridgeman A et al (2012) Superior induction of T cell responses to conserved HIV-1 regions by electroporated alphavirus replicon DNA compared to that with conventional plasmid DNA vaccine. J Virol 86(8):4082–4090

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Liljestrom P, Garoff H (1991) A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Biotechnology 9(12):1356–1361

    Article  CAS  PubMed  Google Scholar 

  • Loy JD, Gander J, Mogler M, Vander Veen R, Ridpath J, Harris DH et al (2013) Development and evaluation of a replicon particle vaccine expressing the E2 glycoprotein of bovine viral diarrhea virus (BVDV) in cattle. Virol J 10:35

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mossman SP, Bex F, Berglund P, Arthos J, O’Neil SP, Riley D et al (1996) Protection against lethal simian immunodeficiency virus SIVsmmPBj14 disease by a recombinant Semliki Forest virus gp160 vaccine and by a gp120 subunit vaccine. J Virol 70(3):1953–1960

    CAS  PubMed Central  PubMed  Google Scholar 

  • Muniz AE (2012) Venezuelan equine encephalitis in a teenager visiting Central America. Pediatr Emerg Care 28(4):372–375

    Article  PubMed  Google Scholar 

  • Nelson EL, Prieto D, Alexander TG, Pushko P, Lofts LA, Rayner JO et al (2003) Venezuelan equine encephalitis replicon immunization overcomes intrinsic tolerance and elicits effective anti-tumor immunity to the ‘self’ tumor-associated antigen, neu in a rat mammary tumor model. Breast Cancer Res Treat 82(3):169–183

    Article  CAS  PubMed  Google Scholar 

  • Nishimoto KP, Laust AK, Wang K, Kamrud KI, Hubby B, Smith JF et al (2007) Restricted and selective tropism of a Venezuelan equine encephalitis virus-derived replicon vector for human dendritic cells. Viral Immunol 20(1):88–104

    Article  CAS  PubMed  Google Scholar 

  • Osada T, Morse MA, Hobeika A, Lyerly HK (2012) Novel recombinant alphaviral and adenoviral vectors for cancer immunotherapy. Semin Oncol 39(3):305–310

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Paredes AM, Brown DT, Rothnagel R, Chiu W, Schoepp RJ, Johnston RE et al (1993) Three-dimensional structure of a membrane-containing virus. Proc Natl Acad Sci U S A 90(19):9095–9099

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Petrakova O, Volkova E, Gorchakov R, Paessler S, Kinney RM, Frolov I (2005) Noncytopathic replication of Venezuelan equine encephalitis virus and eastern equine encephalitis virus replicons in Mammalian cells. J Virol 79(12):7597–7608

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Pijlman GP, Suhrbier A, Khromykh AA (2006) Kunjin virus replicons: an RNA-based, non-cytopathic viral vector system for protein production, vaccine and gene therapy applications. Expert Opin Biol Ther 6(2):135–145

    Article  CAS  PubMed  Google Scholar 

  • Pushko P, Parker M, Ludwig GV, Davis NL, Johnston RE, Smith JF (1997) Replicon-helper systems from attenuated Venezuelan equine encephalitis virus: expression of heterologous genes in vitro and immunization against heterologous pathogens in vivo. Virology 239(2):389–401

    Article  CAS  PubMed  Google Scholar 

  • 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(1):142–153

    Article  CAS  PubMed  Google Scholar 

  • Pushko P, Geisbert J, Parker M, Jahrling P, Smith J (2001) Individual and bivalent vaccines based on alphavirus replicons protect guinea pigs against infection with Lassa and Ebola viruses. J Virol 75(23):11677–11685

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Schafer A, Whitmore AC, Konopka JL, Johnston RE (2009) Replicon particles of Venezuelan equine encephalitis virus as a reductionist murine model for encephalitis. J Virol 83(9):4275–4286

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Schwartz O, Albert ML (2010) Biology and pathogenesis of chikungunya virus. Nat Rev 8(7):491–500

    CAS  Google Scholar 

  • Seregin AV, Yun NE, Poussard AL, Peng BH, Smith JK, Smith JN et al (2010) TC83 replicon vectored vaccine provides protection against Junin virus in guinea pigs. Vaccine 28(30):4713–4718

    Article  CAS  PubMed  Google Scholar 

  • 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(6):943–949

    Article  CAS  PubMed  Google Scholar 

  • Strauss JH, Strauss EG (1994) The alphaviruses: gene expression, replication, and evolution. Microbiol Rev 58(3):491–562

    CAS  PubMed Central  PubMed  Google Scholar 

  • Sun Y, Tian DY, Li S, Meng QL, Zhao BB, Li Y et al (2013) Comprehensive evaluation of the adenovirus/alphavirus-replicon chimeric vector-based vaccine rAdV-SFV-E2 against classical swine fever. Vaccine 31(3):538–544

    Article  CAS  PubMed  Google Scholar 

  • Tegerstedt K, Franzen AV, Andreasson K, Joneberg J, Heidari S, Ramqvist T et al (2005) Murine polyomavirus virus-like particles (VLPs) as vectors for gene and immune therapy and vaccines against viral infections and cancer. Anticancer Res 25(4):2601–2608

    CAS  PubMed  Google Scholar 

  • Thompson JM, Whitmore AC, Konopka JL, Collier ML, Richmond EM, Davis NL et al (2006) Mucosal and systemic adjuvant activity of alphavirus replicon particles. Proc Natl Acad Sci U S A 103(10):3722–3727

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Thompson JM, Whitmore AC, Staats HF, Johnston RE (2008) Alphavirus replicon particles acting as adjuvants promote CD8+ T cell responses to co-delivered antigen. Vaccine 26(33):4267–4275

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Tretyakova I, Lukashevich IS, Glass P, Wang E, Weaver S, Pushko P (2013) Novel vaccine against Venezuelan equine encephalitis combines advantages of DNA immunization and a live attenuated vaccine. Vaccine 31(7):1019–1025

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Vancini R, Wang G, Ferreira D, Hernandez R, Brown DT (2013) Alphavirus genome delivery occurs directly at the plasma membrane in a time- and temperature-dependent process. J Virol 87(8):4352–4359

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Vander Veen RL, Harris DL, Kamrud KI (2012) Alphavirus replicon vaccines. Animal Health Research Reviews/Conference of Research Workers in Animal Diseases 13(1):1–9

    Google Scholar 

  • Vaney MC, Duquerroy S, Rey FA (2013) Alphavirus structure: activation for entry at the target cell surface. Curr Opin Virol 3(2):151–158

    Article  CAS  PubMed  Google Scholar 

  • Vasilakis N, Falvey D, Gangolli SS, Coleman J, Kowalski J, Udem SA et al (2003) Transfection-independent production of alphavirus replicon particles based on poxvirus expression vectors. Nat Biotechnol 21(8):932–935

    Article  CAS  PubMed  Google Scholar 

  • Vasilevska J, Skrastina D, Spunde K, Garoff H, Kozlovska T, Zajakina A (2012) Semliki Forest virus biodistribution in tumor-free and 4T1 mammary tumor-bearing mice: a comparison of transgene delivery by recombinant virus particles and naked RNA replicon. Cancer Gene Ther 19(8):579–587

    Article  CAS  PubMed  Google Scholar 

  • Voss JE, Vaney MC, Duquerroy S, Vonrhein C, Girard-Blanc C, Crublet E et al (2010) Glycoprotein organization of Chikungunya virus particles revealed by X-ray crystallography. Nature 468(7324):709–712

    Article  CAS  PubMed  Google Scholar 

  • Wahlberg JM, Garoff H (1992) Membrane fusion process of Semliki Forest virus. I: Low pH-induced rearrangement in spike protein quaternary structure precedes virus penetration into cells. J Cell Biol 116(2):339–348

    Article  CAS  PubMed  Google Scholar 

  • Wahlberg JM, Boere WA, Garoff H (1989) The heterodimeric association between the membrane proteins of Semliki Forest virus changes its sensitivity to low pH during virus maturation. J Virol 63(12):4991–4997

    CAS  PubMed Central  PubMed  Google Scholar 

  • 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(10):1651–1660

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Werblow A, Bolius S, Dorresteijn AW, Melaun C, Klimpel S (2013) Diversity of Culex torrentium Martini, 1925—a potential vector of arboviruses and filaria in Europe. Parasitol Res 112(7):2495–2501

    Article  PubMed  Google Scholar 

  • White LJ, Sariol CA, Mattocks MD, Wahala MPBW, Yingsiwaphat V, Collier ML et al (2013) An alphavirus vector-based tetravalent dengue vaccine induces a rapid and protective immune response in macaques that differs qualitatively from immunity induced by live virus infection. J Virol 87(6):3409–3424

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Williamson C, Morris L, Maughan MF, Ping LH, Dryga SA, Thomas R et al (2003) Characterization and selection of HIV-1 subtype C isolates for use in vaccine development. AIDS Res Hum Retrovir 19(2):133–144

    Article  CAS  PubMed  Google Scholar 

  • Wolf A, Hodneland K, Frost P, Braaen S, Rimstad E (2013) A hemagglutinin-esterase-expressing salmonid alphavirus replicon protects Atlantic salmon (Salmo salar) against infectious salmon anemia (ISA). Vaccine 31(4):661–669

    Article  CAS  PubMed  Google Scholar 

  • 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 (New York, NY) 243(4895):1188–1191

    Article  CAS  Google Scholar 

  • Yu YZ, Guo JP, An HJ, Zhang SM, Wang S, Yu WY et al (2013) Potent tetravalent replicon vaccines against botulinum neurotoxins using DNA-based Semliki Forest virus replicon vectors. Vaccine 31(20):2427–2432

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Fugere M, Day R, Kielian M (2003) Furin processing and proteolytic activation of Semliki Forest virus. J Virol 77(5):2981–2989

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhang R, Hryc CF, Cong Y, Liu X, Jakana J, Gorchakov R et al (2011) 4.4 A cryo-EM structure of an enveloped alphavirus Venezuelan equine encephalitis virus. EMBO J 30(18):3854–3863

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhou X, Berglund P, Zhao H, Liljestrom P, Jondal M (1995) Generation of cytotoxic and humoral immune responses by nonreplicative recombinant Semliki Forest virus. Proc Natl Acad Sci U S A 92(7):3009–3013

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhu W, Fu J, Lu J, Deng Y, Wang H, Wei Y et al (2013) Induction of humoral and cellular immune responses against hepatitis C virus by vaccination with replicon particles derived from Sindbis-like virus XJ-160. Arch Virol 158(5):1013–1019

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Authors thank Rachmat Hidajat and Igor Lukashevich for valuable contributions and discussions. This publication was supported in part by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under grants R43AI094700 and R01AI093450. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding agency.

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  1. Medigen Inc., Riverside Technology Park, 8420 Gas House Pike, Suite S, Frederick, MD, 21701, USA

    Peter Pushko & Irina Tretyakova

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Correspondence to Peter Pushko .

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  1. Department of Pharmacology and Toxicolog Department of Microbiology and Immunolog, University of Louisville, Louisville, Kentucky, USA

    Igor S Lukashevich

  2. Department of Microbiology and Immunolog, University of Louisville, Louisville, Kentucky, USA

    Haval Shirwan

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Pushko, P., Tretyakova, I. (2014). Alphavirus Replicon Vectors for Prophylactic Applications and Cancer Intervention. In: Lukashevich, I., Shirwan, H. (eds) Novel Technologies for Vaccine Development. Springer, Vienna. https://doi.org/10.1007/978-3-7091-1818-4_3

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