Brazilian Journal of Microbiology

, Volume 50, Issue 2, pp 347–355 | Cite as

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


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.


MVA-based vaccine Green fluorescent protein GFP Acute responses 



Alanine aminotransferase


Aspartate aminotransferase


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


Vaccinia virus



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.


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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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