Skip to main content
Top
Published in: Virology Journal 1/2023

Open Access 01-12-2023 | Methodology

Generation of a recombinant Saffold Virus expressing UnaG as a marker for the visualization of viral infection

Authors: Takako Okuwa, Toshiki Himeda, Koichi Utani, Masaya Higuchi

Published in: Virology Journal | Issue 1/2023

Login to get access

Abstract

Background

Saffold virus (SAFV), which belongs to the genus Cardiovirus of the family Picornaviridae, is associated with acute respiratory or gastrointestinal illnesses in children; it is also suspected to cause severe diseases, such as acute flaccid paralysis and aseptic meningitis. However, the understanding of the mechanism of its pathogenicity is still limited due to the many unknowns about its lifecycle; for example, the cellular receptor for its infection remains to be determined. A system to monitor SAFV infection in vitro and in vivo is required in order to accelerate research on SAFV.

Results

We generated a recombinant SAFV expressing green fluorescent protein (GFP) or UnaG, a novel fluorescent protein derived from Japanese eel. HeLa cells infected by either GFP or UnaG-expressing SAFV showed a bright green fluorescent signal, enabling convenient monitoring of SAFV infection. However, the expression of GFP but not UnaG was quickly lost during virus passaging due to the difference in genetic stability in the SAFV virus genome; the UnaG gene was stably maintained in the virus genome after at least five passages.

Conclusions

SAFV infection of cultured cells can easily be monitored using UnaG-expressing SAFV, which is superior to GFP in terms of genetic stability in the virus genome. This virus could be a useful tool for SAFV research, such as comparing the susceptibility of various cells to SAFV infection and evaluating the effects of antivirals on SAFV infection in high-throughput screening.
Appendix
Available only for authorised users
Literature
1.
go back to reference Jones MS, Lukashov VV, Ganac RD, Schnurr DP. Discovery of a novel human picornavirus in a stool sample from a pediatric patient presenting with fever of unknown origin. J Clin Microbiol. 2007;45:2144–50.CrossRefPubMedPubMedCentral Jones MS, Lukashov VV, Ganac RD, Schnurr DP. Discovery of a novel human picornavirus in a stool sample from a pediatric patient presenting with fever of unknown origin. J Clin Microbiol. 2007;45:2144–50.CrossRefPubMedPubMedCentral
3.
go back to reference Lin TL, Lin TH, Chiu SC, Huang YP, Ho CM, Lee CC, Wu HS, Lin JH. Molecular epidemiological analysis of Saffold cardiovirus genotype 3 from upper respiratory infection patients in Taiwan. J Clin Virol. 2015;70:7–13.CrossRefPubMed Lin TL, Lin TH, Chiu SC, Huang YP, Ho CM, Lee CC, Wu HS, Lin JH. Molecular epidemiological analysis of Saffold cardiovirus genotype 3 from upper respiratory infection patients in Taiwan. J Clin Virol. 2015;70:7–13.CrossRefPubMed
4.
go back to reference Itagaki T, Aoki Y, Matoba Y, Tanaka S, Ikeda T, Matsuzaki Y, Mizuta K. Detection of Saffold viruses from children with acute respiratory infections in Yamagata, Japan, between 2008 and 2015. J Med Virol. 2018;90:34–40.CrossRefPubMed Itagaki T, Aoki Y, Matoba Y, Tanaka S, Ikeda T, Matsuzaki Y, Mizuta K. Detection of Saffold viruses from children with acute respiratory infections in Yamagata, Japan, between 2008 and 2015. J Med Virol. 2018;90:34–40.CrossRefPubMed
5.
go back to reference Drexler JF, Luna LKS, Stöcker A, Almeida PS, Ribeiro TCM, Petersen N, Herzog P, Pedroso C, Huppertz HI, Ribeiro HD, et al. Circulation of 3 lineages of a novel Saffold cardiovirus in humans. Emerg Infect Dis. 2008;14:1398–405.CrossRefPubMedPubMedCentral Drexler JF, Luna LKS, Stöcker A, Almeida PS, Ribeiro TCM, Petersen N, Herzog P, Pedroso C, Huppertz HI, Ribeiro HD, et al. Circulation of 3 lineages of a novel Saffold cardiovirus in humans. Emerg Infect Dis. 2008;14:1398–405.CrossRefPubMedPubMedCentral
6.
go back to reference Nielsen ACY, Gyhrs ML, Nielsen LP, Pedersen C, Böttiger B. Gastroenteritis and the novel picornaviruses aichi virus, cosavirus, saffold virus, and salivirus in young children. J Clin Virol. 2013;57:239–42.CrossRefPubMed Nielsen ACY, Gyhrs ML, Nielsen LP, Pedersen C, Böttiger B. Gastroenteritis and the novel picornaviruses aichi virus, cosavirus, saffold virus, and salivirus in young children. J Clin Virol. 2013;57:239–42.CrossRefPubMed
7.
go back to reference Naeem A, Hosomi T, Nishimura Y, Alam MM, Oka T, Zaidi SSZ, Shimizu H. Genetic diversity of circulating Saffold viruses in Pakistan and Afghanistan. J Gen Virol. 2014;95:1945–57.CrossRefPubMed Naeem A, Hosomi T, Nishimura Y, Alam MM, Oka T, Zaidi SSZ, Shimizu H. Genetic diversity of circulating Saffold viruses in Pakistan and Afghanistan. J Gen Virol. 2014;95:1945–57.CrossRefPubMed
8.
go back to reference Himeda T, Hosomi T, Asif N, Shimizu H, Okuwa T, Muraki Y, Ohara Y. The preparation of an infectious full-length cDNA clone of Saffold virus. Virol J 2011;8:110. Himeda T, Hosomi T, Asif N, Shimizu H, Okuwa T, Muraki Y, Ohara Y. The preparation of an infectious full-length cDNA clone of Saffold virus. Virol J 2011;8:110.
9.
go back to reference Nielsen TS, Nielsen AY, Banner J, Hansen J, Baandrup U, Nielsen LP. Saffold virus infection associated with human myocarditis. J Clin Virol. 2016;74:78–81.CrossRefPubMed Nielsen TS, Nielsen AY, Banner J, Hansen J, Baandrup U, Nielsen LP. Saffold virus infection associated with human myocarditis. J Clin Virol. 2016;74:78–81.CrossRefPubMed
10.
go back to reference Ito H, Miyagaki S, Sakaue S, Matsui F, Katsumi Y, Otabe O, Torii J, Itagaki T, Himeda T, Okuwa T, Ohara Y. Saffold cardiovirus infection in a 2-year-old boy with acute pancreatitis. Jpn J Infect Dis. 2017;70:105–07.CrossRefPubMed Ito H, Miyagaki S, Sakaue S, Matsui F, Katsumi Y, Otabe O, Torii J, Itagaki T, Himeda T, Okuwa T, Ohara Y. Saffold cardiovirus infection in a 2-year-old boy with acute pancreatitis. Jpn J Infect Dis. 2017;70:105–07.CrossRefPubMed
11.
go back to reference Nielsen ACY, Böttiger B, Banner J, Hoffmann T, Nielsen LP. Serious invasive Saffold virus infections in children, 2009. Emerg Infect Dis. 2012;18:7–12.CrossRefPubMedPubMedCentral Nielsen ACY, Böttiger B, Banner J, Hoffmann T, Nielsen LP. Serious invasive Saffold virus infections in children, 2009. Emerg Infect Dis. 2012;18:7–12.CrossRefPubMedPubMedCentral
13.
go back to reference Martinez-Salas E, Ryan MD. Translation and protein processing. In: Ehrenfeld E, Domingo E, Roos RP, editors. The picornaviruses. Washington, DC: ASM Press; 2010. pp. 141–61. Martinez-Salas E, Ryan MD. Translation and protein processing. In: Ehrenfeld E, Domingo E, Roos RP, editors. The picornaviruses. Washington, DC: ASM Press; 2010. pp. 141–61.
14.
go back to reference Luke GA, de Felipe P, Lukashev A, Kallioinen SE, Bruno EA, Ryan MD. Occurrence, function and evolutionary origins of ‘2A-like’ sequences in virus genomes. J Gen Virol. 2008;89:1036–42.CrossRefPubMedPubMedCentral Luke GA, de Felipe P, Lukashev A, Kallioinen SE, Bruno EA, Ryan MD. Occurrence, function and evolutionary origins of ‘2A-like’ sequences in virus genomes. J Gen Virol. 2008;89:1036–42.CrossRefPubMedPubMedCentral
15.
go back to reference Kumagai A, Ando R, Miyatake H, Greimel P, Kobayashi T, Hirabayashi Y, Shimogori T, Miyawaki A. A bilirubin-inducible fluorescent protein from eel muscle. Cell. 2013;153:1602–11.CrossRefPubMed Kumagai A, Ando R, Miyatake H, Greimel P, Kobayashi T, Hirabayashi Y, Shimogori T, Miyawaki A. A bilirubin-inducible fluorescent protein from eel muscle. Cell. 2013;153:1602–11.CrossRefPubMed
16.
go back to reference Finch LK, Ling R, Napthine S, Olspert A, Michiels T, Lardinois C, Bell S, Loughran G, Brierley I, Firth AE. Characterization of ribosomal frameshifting in Theiler’s murine encephalomyelitis virus. J Virol. 2015;89:8580–89.CrossRefPubMedPubMedCentral Finch LK, Ling R, Napthine S, Olspert A, Michiels T, Lardinois C, Bell S, Loughran G, Brierley I, Firth AE. Characterization of ribosomal frameshifting in Theiler’s murine encephalomyelitis virus. J Virol. 2015;89:8580–89.CrossRefPubMedPubMedCentral
17.
go back to reference Loughran G, Firth AE, Atkins JF. Ribosomal frameshifting into an overlapping gene in the 2B-encoding region of the cardiovirus genome. Proc Natl Acad Sci U S A. 2011;108:E1111–9.CrossRefPubMedPubMedCentral Loughran G, Firth AE, Atkins JF. Ribosomal frameshifting into an overlapping gene in the 2B-encoding region of the cardiovirus genome. Proc Natl Acad Sci U S A. 2011;108:E1111–9.CrossRefPubMedPubMedCentral
18.
go back to reference Binder JJ, Hoffman MA, Palmenberg AC. Genetic stability of attenuated mengovirus vectors with duplicate primary cleavage sequences. Virology. 2003;312:481–94.CrossRefPubMed Binder JJ, Hoffman MA, Palmenberg AC. Genetic stability of attenuated mengovirus vectors with duplicate primary cleavage sequences. Virology. 2003;312:481–94.CrossRefPubMed
19.
go back to reference Zoll J, Hulshof SE, Lanke K, Lunel FV, Melchers WJG, Schoondermark-van de Ven E, Roivainen M, Galama JMD, van Kuppeveld FJM. Saffold virus, a human Theiler’s-like cardiovirus, is ubiquitous and causes infection early in life. PLoS Pathog 2009; 5:e1000416. Zoll J, Hulshof SE, Lanke K, Lunel FV, Melchers WJG, Schoondermark-van de Ven E, Roivainen M, Galama JMD, van Kuppeveld FJM. Saffold virus, a human Theiler’s-like cardiovirus, is ubiquitous and causes infection early in life. PLoS Pathog 2009; 5:e1000416.
20.
go back to reference Himeda T, Hosomi T, Okuwa T, Muraki Y, Ohara Y. Saffold virus type 3 (SAFV-3) persists in HeLa cells. PLoS ONE 2013; 8:e53194. Himeda T, Hosomi T, Okuwa T, Muraki Y, Ohara Y. Saffold virus type 3 (SAFV-3) persists in HeLa cells. PLoS ONE 2013; 8:e53194.
22.
go back to reference Mullapudi E, Nováček J, Pálková L, Kulich P, Lindberg AM, van Kuppeveld FJM, Plevka P. Structure and genome release mechanism of the human cardiovirus Saffold Virus 3. J Virol. 2016;90:7628–39.CrossRefPubMedPubMedCentral Mullapudi E, Nováček J, Pálková L, Kulich P, Lindberg AM, van Kuppeveld FJM, Plevka P. Structure and genome release mechanism of the human cardiovirus Saffold Virus 3. J Virol. 2016;90:7628–39.CrossRefPubMedPubMedCentral
23.
go back to reference Philip AA, Perry JL, Eaton HE, Shmulevitz M, Hyser JM, Patton JT. Generation of recombinant rotavirus expressing NSP3-UnaG fusion protein by a simplified reverse genetics system. J Virol 2019; 93:e01616-19. Philip AA, Perry JL, Eaton HE, Shmulevitz M, Hyser JM, Patton JT. Generation of recombinant rotavirus expressing NSP3-UnaG fusion protein by a simplified reverse genetics system. J Virol 2019; 93:e01616-19.
24.
go back to reference Liu Y, Zhang H, Chen R, Wu Y, Yang X, Liu X, Zeng S, Guo W. UnaG as a reporter in adeno-associated virus‐mediated gene transfer for biomedical imaging. J Biophotonics. 2020;13:e202000182.CrossRefPubMed Liu Y, Zhang H, Chen R, Wu Y, Yang X, Liu X, Zeng S, Guo W. UnaG as a reporter in adeno-associated virus‐mediated gene transfer for biomedical imaging. J Biophotonics. 2020;13:e202000182.CrossRefPubMed
Metadata
Title
Generation of a recombinant Saffold Virus expressing UnaG as a marker for the visualization of viral infection
Authors
Takako Okuwa
Toshiki Himeda
Koichi Utani
Masaya Higuchi
Publication date
01-12-2023
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2023
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/s12985-023-02142-8

Other articles of this Issue 1/2023

Virology Journal 1/2023 Go to the issue
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.