Vaccinia Virus Expression System

Abstract

Replication‐competent vaccinia virus was used as the vaccine in the campaign to eradicate smallpox. After the disease was eradicated, recombinant vaccinia viruses were created that were capable of expressing foreign genes. This made vaccinia‐based systems available for mammalian cell expression of recombinant protein, an advantage of which, like other mammalian expression systems, is authentic posttranslational modification. Generation of recombinant vaccinia also provided an opportunity for developing vaccinia‐based vaccines for use against nonpoxvirus diseases. In this respect, recombinant vaccinia virus and recombinant versions of an attenuated derivative, Modified Vaccinia virus Ankara (MVA) have been extensively studied. Today, there are recombinant vaccinia viruses expressing foreign antigens in phases I–III clinical trials relating to viral, bacterial, parasitic and cancer‐related diseases, and a licensed veterinary vaccine.

Key Concepts:

  • Smallpox was eradicated following a vaccination campaign in which live vaccinia virus was used as the vaccine.

  • Foreign genes can be inserted into the vaccinia virus genome to create recombinant vaccinia virus strains.

  • The vaccinia virus expression system allows for mammalian cell expression of recombinant protein, an advantage of which is authentic post‐translational modification.

  • Recombinant vaccinia can deliver vaccine antigens and evoke potent cell‐mediated and humoral responses specific to the inserted gene.

  • Vaccination with recombinant organisms can confer protection against viral, bacterial, parasitic and cancer‐related diseases.

Keywords: vaccinia virus; recombinant virus; expression system; viral vector; vaccine

Figure 1.

Overview of the vaccinia virus life cycle. Four types of vaccinia virion are formed during each round of replication. Intracellular mature virus (IMV) is released on cell lysis. Cell‐associated enveloped virus (CEV) is responsible for cell‐to‐cell spread and extracellular enveloped virus (EEV) for longer range dissemination. The intracellular enveloped virus (IEV) represents an IMV–CEV/EEV intermediate (Smith et al., ). Adapted from Hall and Carroll (). © Wiley‐Blackwell.

Figure 2.

Immunostained plaques. BHK‐21 and CEF monolayers were infected with MVA. After 24 h, cells were fixed and immunostained using anti‐vaccinia antibody to visualise infected cells.

Figure 3.

Typical features of a transfer plasmid. Left and right flanks homologous to the vaccinia genome determine the point and orientation of insertion for the recombinant cassette. The gene of interest (GOI) and a selection marker, green fluorescent protein (GFP) are inserted, each with its own vaccinia promoter (P) for control of gene expression.

Figure 4.

Detection of recombinant vaccinia expression. CEF cell lysate samples were separated by SDS PAGE, transferred to nitrocellulose via Western Blot and then subjected to immunodetection using antibody specific for the recombinant insert. Each sample had been infected with a different recombinant MVA (rMVA) strain: (1) rMVA expressing gene A (48 kDa); (2) rMVA expressing gene B (83 kDa) and (3) multiple‐rMVA expressing two inserts, gene A and gene B.

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References

Antoine G, Scheiflinger F, Dorner F and Falkner FG (1998) The complete genomic sequence of the modified vaccinia Ankara strain: comparison with other orthopoxviruses. Virology 244: 365–396.

Baldick CJ Jr. and Moss B (1993) Characterization and temporal regulation of mRNAs encoded by vaccinia virus intermediate‐stage genes. Journal of Virology 67: 3515–3527.

Bennink JR, Yewdell JW, Smith GL, Moller C and Moss B (1984) Recombinant vaccinia virus primes and stimulates influenza haemagglutinin‐specific cytotoxic T cells. Nature 311: 578–579.

Broyles SS (2003) Vaccinia virus transcription. Journal of General Virology 84: 2293–2303.

Carroll MW and Moss B (1997) Host range and cytopathogenicity of the highly attenuated MVA strain of vaccinia virus: propagation and generation of recombinant viruses in a nonhuman mammalian cell line. Virology 238: 198–211.

Chakrabarti S, Brechling K and Moss B (1985) Vaccinia virus expression vector: coexpression of beta‐galactosidase provides visual screening of recombinant virus plaques. Molecular and Cellular Biology 5: 3403–3409.

Chakrabarti S, Sisler JR and Moss B (1997) Compact, synthetic, vaccinia virus early/late promoter for protein expression. Biotechniques 23: 1094–1097.

Davison AJ and Moss B (1989a) Structure of vaccinia virus early promoters. Journal of Molecular Biology 210: 749–769.

Davison AJ and Moss B (1989b) Structure of vaccinia virus late promoters. Journal of Molecular Biology 210: 771–784.

Domi A and Moss B (2002) Cloning the vaccinia virus genome as a bacterial artificial chromosome in Escherichia coli and recovery of infectious virus in mammalian cells. Proceedings of the National Academy of Sciences of the USA 99: 12415–12420.

Dreicer R, Stadler WM, Ahmann FR et al. (2009) MVA‐MUC1‐IL2 vaccine immunotherapy (TG4010) improves PSA doubling time in patients with prostate cancer with biochemical failure. Investigational New Drugs 27: 379–386.

Earl PL, Hugin AW and Moss B (1990) Removal of cryptic poxvirus transcription termination signals from the human immunodeficiency virus type 1 envelope gene enhances expression and immunogenicity of a recombinant vaccinia virus. Journal of Virology 64: 2448–2451.

Fuerst TR, Niles EG, Studier FW and Moss B (1986) Eukaryotic transient‐expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proceedings of the National Academy of Sciences of the USA 83: 8122–8126.

Garcia‐Arriaza J, Arnaez P, Gomez CE, Sorzano CO and Esteban M (2013) Improving adaptive and memory immune responses of an HIV/AIDS vaccine candidate MVA‐B by deletion of vaccinia virus genes (C6L and K7R) blocking interferon signaling pathways. PLoS One 8: e66894.

Hall Y and Carroll MW (2012) Recombinant MVA vaccines: optimization, preclinical, and product development. In: Morrow WJW, Sheikh NA, Schmidt CS, Davies HD (eds) Vaccinology Principles and Practice, pp. 209–223. Chichester: Wiley‐Blackwell.

Lane JM, Ruben FL, Neff JM and Millar JD (1969) Complications of smallpox vaccination, 1968. New England Journal of Medicine 281: 1201–1208.

Lohr V, Rath A, Genzel Y et al. (2009) New avian suspension cell lines provide production of influenza virus and MVA in serum‐free media: studies on growth, metabolism and virus propagation. Vaccine 27: 4975–4982.

Mackett M, Smith GL and Moss B (1982) Vaccinia virus: a selectable eukaryotic cloning and expression vector. Proceedings of the National Academy of Sciences of the USA 79: 7415–7419.

Mayr A, Stickl H, Muller HK, Danner K and Singer H (1978) The smallpox vaccination strain MVA: marker, genetic structure, experience gained with the parenteral vaccination and behavior in organisms with a debilitated defence mechanism (author's transl). Zentralbl Bakteriol B 167: 375–390.

Merchlinsky M and Moss B (1992). Introduction of foreign DNA into the vaccinia virus genome by in vitro ligation: recombination‐independent selectable cloning vectors. Virology 190: 522–526.

Perkus ME, Limbach K and Paoletti E (1989) Cloning and expression of foreign genes in vaccinia virus, using a host range selection system. Journal of Virology 63: 3829–3836.

Smith GL, Vanderplasschen A and Law M (2002) The formation and function of extracellular enveloped vaccinia virus. Journal of General Virology 83: 2915–2931.

Sutter G and Moss B (1992) Nonreplicating vaccinia vector efficiently expresses recombinant genes. Proceedings of the National Academy of Sciences of the USA 89: 10847–10851.

Sutter G, Wyatt LS, Foley PL, Bennink JR and Moss B (1994) A recombinant vector derived from the host range‐restricted and highly attenuated MVA strain of vaccinia virus stimulates protective immunity in mice to influenza virus. Vaccine 12: 1032–1040.

Timm A, Enzinger C, Felder E and Chaplin P (2006) Genetic stability of recombinant MVA‐BN. Vaccine 24: 4618–4621.

Walker BD, Flexner C, Paradis TJ et al. (1988) HIV‐1 reverse transcriptase is a target for cytotoxic T lymphocytes in infected individuals. Science 240: 64–66.

Wyatt LS, Moss B and Rozenblatt S (1995) Replication‐deficient vaccinia virus encoding bacteriophage T7 RNA polymerase for transient gene expression in mammalian cells. Virology 210: 202–205.

Further Reading

Earl PL, Cooper N, Wyatt LS, Moss B and Carroll MW (2001a) Preparation of cell cultures and vaccinia virus stocks. Current Protocols in Protein Science, Chapter 5, Unit5 12.

Earl PL, Moss B, Wyatt LS and Carroll MW (2001b) Generation of recombinant vaccinia viruses. Current Protocols in Protein Science, Chapter 5, Unit5 13.

Earl PL and Moss B (2001) Characterization of recombinant vaccinia viruses and their products. Current Protocols in Protein Science, Chapter 5, Unit5 14.

Moss B (2007) Poxviridae: the viruses and their replication. In: Fields BN, Knipe DM, Howley PM (eds) Fields Virology, pp. 2905–2946. Philadelphia: Lippincott‐Raven Publishers.

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Hall, Yper, and Carroll, Miles W(May 2014) Vaccinia Virus Expression System. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002659.pub3]