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The Perennial Use of the Green Fluorescent Protein Marker in a Live Vaccinia Virus Ankara Recombinant Platform Shows No Acute Adverse Effects in Mice

  • D. S. O. Daian e Silva
  • T. M. G. Pinho
  • M. A. Rachid
  • D. F. Barbosa-Stancioli
  • F. G. Da FonsecaEmail author
Bacterial, Fungal and Virus Molecular Biology - Research Paper

Abstract

Recombinant virus vectors represent a promising strategy for vaccine research. Among available viral vectors, members of the Poxviridae family—especially the modified Vaccinia virus Ankara (MVA)—stand out as immunogenic and safe vaccine platforms. Because MVA usually does not produce plaques in cell culture, visible selection markers such as the green fluorescent protein (GFP) are frequently incorporated into the constructions in order to facilitate the recognition of recombinants. However, these genetic markers have to be removed before any clinical trial. Here, we evaluated the acute responses generated in mice immunized with a MVA vector in which the GFP marker was not removed. We observed no differences in neutrophil, monocyte, or total leucocyte recruitment among animals inoculated with MVA or MVA-GFP. Likewise, there were no differences in neutrophil activation between mice groups. Hepatic functions were not altered in either MVA or MVA-GFP-inoculated mice, and we observed no histopathological alterations in different tissues from virus-inoculated animals. In conclusion, the presence of GFP is innocuous to immunized animals and do not alter acute physiopathological responses to the MVA vector. We suggest that keeping the GFP marker may be a good strategy for vaccine development, production, and evaluation.

Keywords

MVA-based vaccine Green fluorescent protein GFP Acute responses 

Abbreviations

ALT

Alanine aminotransferase

AST

Aspartate aminotransferase

CEFs

Chicken embryo fibroblast cells

GFP

Green fluorescente protein

MPO

Myeloperoxidase

MVA

Wild-type modified Vaccinia virus Ankara

MVA-GFP

Modified Vaccinia virus Ankara-expressing GFP protein

MOI

Multiplicity of infection

PFU

Plaque forming unit

VACV

Vaccinia virus

Notes

Acknowledgments

We are grateful to Prof. Daniele da Glória de Souza and their team for their help in evaluating the data and for critical advice. MA Rachid and EF Barbosa-Stancioli are CNPq Fellowship Recipients.

Funding information

This research was supported by CAPES and the Post-Graduation program in Microbiology from the Universidade Federal de Minas Gerais. Financial resources came also from grants by FAPEMIG and CNPq.

Compliance with ethical standards

All procedures reported here are in accordance with the ethical principles of animal experimentation adopted by the Ethics Committee for Animal experiments from Universidade Federal de Minas Gerais (CETEA/UFMG—protocol 273/2008).

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Luyten J, Beutels P (2016) The social value of vaccination programs: beyond cost-effectiveness. Health Aff (Millwood) 35:212–218CrossRefGoogle Scholar
  2. 2.
    Buckland BC (2005) The process development challenge for a new vaccine. Nat Med 11:16–19CrossRefGoogle Scholar
  3. 3.
    Plotkin SA, Mahmoud AAF, Farrar J (2015) Establishing a global vaccine-development fund. N Engl J Med 373:297–300CrossRefPubMedGoogle Scholar
  4. 4.
    Andrade LM, Cox L, Versiani AF, da Fonseca FG (2017) A growing world of small things: a brief review on the nanostructured vaccines. Futur Virol 12:767–779CrossRefGoogle Scholar
  5. 5.
    Moss B (2007) Poxviridae: the viruses and their replication. In: Fields Virology 2, 5th edn. Lippincott Williams & Wilkins, Philadelphia, pp 2637–2267Google Scholar
  6. 6.
    Moss B (2011) Smallpox vaccines: targets of protective immunity. Immunol Rev 239:8–26CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Quinan BR, Daian e Silva DSO, Coelho FM, Da Fonseca FG (2014) Modified vaccinia virus Ankara as vaccine vectors in human and veterinary medicine. Futur Virol 9:173–187CrossRefGoogle Scholar
  8. 8.
    Drexler I, Staib C, Sutter G (2004) Modified vaccinia virus Ankara as antigen delivery system: how can we best use its potential? Curr Opin Biotechnol 15:506–512CrossRefPubMedGoogle Scholar
  9. 9.
    Moss B (2013) Reflections on the early development of poxvirus vectors. Vaccine. 31:4220–4222CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Zuverink M, Barbieri JT (2015) From GFP to β-lactamase: advancing intact cell imaging for toxins and effectors. Pathog Dis 73:ftv097CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Bisht H, Roberts A, Vogel L, Bukreyev A, Collins PL, Murphy BR, Subbarao K, Moss B (2004) Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice. Proc Natl Acad Sci U S A 101:6641–6646CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Souza DG, Cara DC, Cassali GD, Coutinho SF, Silveira MR, Andrade SP, Poole SP, Teixeira MM (2000) Effects of the PAF receptor antagonist UK74505 on local and remote reperfusion injuries following ischaemia of the superior mesenteric artery in the rat. Br J Pharmacol 131:1800–1808CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Barcelos LS, Talvani A, Teixeira AS, Cassali GD, Andrade SP, Teixeira MM (2004) Production and in vivo effects of chemokines CXCL1-3/KC and CCL2/JE in a model of inflammatory angiogenesis in mice. Inflamm Res 53:576–584CrossRefPubMedGoogle Scholar
  14. 14.
    Ogawa H, Inouye S, Tsuji FI, Yasuda K, Umesono K (1995) Localization, trafficking, and temperature-dependence of the Aequorea green fluorescent protein in cultured vertebrate cells. Proc Natl Acad Sci U S A 92:11899–11903CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Ramirez JC, Finke D, Esteban M, Kraehenbuhl JP, Acha-Orbea H (2003) Tissue distribution of the Ankara strain of vaccinia virus (MVA) after mucosal or systemic administration. Arch Virol 148:827–839CrossRefPubMedGoogle Scholar
  16. 16.
    van der Veen BS, de Winther MP, Heeringa P (2009) Myeloperoxidase: molecular mechanisms of action and their relevance to human health and disease. Antioxid Redox Signal 11:2899–2937CrossRefPubMedGoogle Scholar
  17. 17.
    Bekheet IW, Madkour ME, Ghaffar NA, Nosseir MMF, Moussa MM, Ibraheim RA, Ateya ME (2009) The role of myeloperoxidase in hepatitis C virus infection and associated liver cirrhosis. Open Trop Med J 2:1–7CrossRefGoogle Scholar
  18. 18.
    Kothari N, Keshari RS, Bogra J, Kohli M, Abbas H, Malik A, Dikshit M, Barthwal MK (2011) Increased myeloperoxidase enzyme activity in plasma is an indicator of inflammation and onset of sepsis. J Crit Care 26:435–e1-435-e.7CrossRefPubMedGoogle Scholar
  19. 19.
    Prokopowicz Z, Marcinkiewicz J, Katz DR, Chain BM (2012) Neutrophil myeloperoxidase: soldier and statesman. Arch Immunol Ther Exp 60:43–54CrossRefGoogle Scholar
  20. 20.
    Klebanoff SJ (2005) Myeloperoxidase: friend and foe. J Leukoc Biol 77:598–625CrossRefPubMedGoogle Scholar
  21. 21.
    Edwards L, Wanless IR (2013) Mechanisms of liver involvement in systemic disease. Best Pract Res Clin Gastroenterol 27:471–483CrossRefPubMedGoogle Scholar
  22. 22.
    Campos F, Rodríguez-Yáñez M, Castellanos M, Arias S, Pérez-Mato M, Sobrino T, Blanco M, Serena J, Castillo J (2011) Blood levels of glutamate oxaloacetate transaminase are more strongly associated with good outcome in acute ischaemic stroke than glutamate pyruvate transaminase levels. Clin Sci 121:11–17CrossRefPubMedGoogle Scholar
  23. 23.
    Ozer J, Ratner M, Shaw M, Bailey W, Schomaker S (2008) The current state of serum biomarkers of hepatotoxicity. Toxicology. 245:194–205CrossRefPubMedGoogle Scholar
  24. 24.
    Earl PL, Americo JL, Moss B (2017) Insufficient innate immunity contributes to the susceptibility of the castaneous mouse to orthopoxvirus infection. J Virol 91:e01042–e01017CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Dowall SD, Graham VA, Rayner E, Hunter L, Watson R, Taylor I et al (2016) Protective effects of a Modified Vaccinia Ankara-based vaccine candidate against Crimean-Congo Haemorrhagic Fever virus require both cellular and humoral responses. PLoS One 11(6):e0156637CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Langenmayer MC, Lülf-Averhoff AT, Adam-Neumair S, Fux R, Sutter G, Volz A (2018) Distribution and absence of generalized lesions in mice following single dose intramuscular inoculation of the vaccine candidate MVA-MERS-S. Biologicals. 54:58–62CrossRefPubMedGoogle Scholar
  27. 27.
    Domínguez LE, Brandmüller C, Zorn J, Kirschning CJ, Sutter G, Lehmann MH et al (2014) Chemokine (C-C Motif) receptor 1 is required for efficient recruitment of neutrophils during respiratory infection with modified vaccinia virus Ankara. J Virol 88:10840–10850CrossRefGoogle Scholar
  28. 28.
    Price PJ, Bánki Z, Scheideler A, Stoiber H, Verschoor A, Sutter G et al (2015) Complement component C5 recruits neutrophils in the absence of C3 during respiratory infection with modified vaccinia virus Ankara. J Immunol 194:1164–1168CrossRefPubMedGoogle Scholar

Copyright information

© Sociedade Brasileira de Microbiologia 2019

Authors and Affiliations

  • D. S. O. Daian e Silva
    • 1
  • T. M. G. Pinho
    • 1
  • M. A. Rachid
    • 2
  • D. F. Barbosa-Stancioli
    • 1
  • F. G. Da Fonseca
    • 1
    Email author
  1. 1.Laboratory of Basic and Applied Virology, Departmento de Microbiologia, Instituto de Ciências BiológicasUniversidade Federal de Minas GeraisBelo HorizonteBrazil
  2. 2.Departamento de Patologia, Instituto de Ciências BiológicasUniversidade Federal de Minas GeraisBelo HorizonteBrazil

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