The Perennial Use of the Green Fluorescent Protein Marker in a Live Vaccinia Virus Ankara Recombinant Platform Shows No Acute Adverse Effects in Mice
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.
KeywordsMVA-based vaccine Green fluorescent protein GFP Acute responses
Chicken embryo fibroblast cells
Green fluorescente protein
Wild-type modified Vaccinia virus Ankara
Modified Vaccinia virus Ankara-expressing GFP protein
Multiplicity of infection
Plaque forming unit
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.
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.
- 5.Moss B (2007) Poxviridae: the viruses and their replication. In: Fields Virology 2, 5th edn. Lippincott Williams & Wilkins, Philadelphia, pp 2637–2267Google Scholar
- 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
- 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
- 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