Bioluminescence imaging of vaccinia virus

Effects of interferon on viral replication and spread

Kathryn E. Luker, Martha Hutchens, Tracey Schultz, Andrew Pekosz, Gary D. Luker

Research output: Contribution to journalArticle

Abstract

Whole animal imaging allows viral replication and localization to be monitored in intact animals, which provides significant advantages for determining viral and host factors that determine pathogenesis. To investigate effects of interferons on spatial and temporal progression of vaccinia infection, we generated recombinant viruses that express firefly luciferase or a monomeric orange fluorescent protein. These viruses allow vaccinia infection to be monitored with bioluminescence or fluorescence imaging, respectively. The recombinant viruses were not attenuated in vitro or in vivo relative to a control WR virus. In cell culture, reporters could be detected readily by 4 h post-infection, showing that these viruses can be used as early markers of infection. The magnitude of firefly luciferase activity measured with bioluminescence imaging in vitro and in vivo correlated directly with increasing titers of vaccinia virus, validating imaging data as a marker of viral infection. Replication of vaccinia was significantly greater in mice lacking receptors for type I interferons (IFN I R-/-) compared with wild-type mice, although both genotypes of mice developed focal infections in lungs and brain after intranasal inoculation. IFN I R-/- mice had greater dissemination of virus to liver and spleen than wild-type animals even when mortality occurred at the same time point after infection. Protective effects of type I interferons were mediated primarily through parenchymal cells rather than hematopoietic cells as analyzed by bone marrow transplant experiments. Collectively, our data define a new function for type I interferon signaling in systemic dissemination of vaccinia and validate these reporter viruses for studies of pathogenesis.

Original languageEnglish (US)
Pages (from-to)284-300
Number of pages17
JournalVirology
Volume341
Issue number2
DOIs
StatePublished - Oct 25 2005
Externally publishedYes

Fingerprint

Vaccinia virus
Interferons
Vaccinia
Viruses
Firefly Luciferases
Interferon Type I
Virus Diseases
Infection
Interferon alpha-beta Receptor
Focal Infection
Wild Animals
Optical Imaging
Spleen
Cell Culture Techniques
Bone Marrow
Genotype
Transplants
Lung
Mortality
Liver

Keywords

  • Bioluminescence imaging
  • Interferon
  • Vaccinia

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Bioluminescence imaging of vaccinia virus : Effects of interferon on viral replication and spread. / Luker, Kathryn E.; Hutchens, Martha; Schultz, Tracey; Pekosz, Andrew; Luker, Gary D.

In: Virology, Vol. 341, No. 2, 25.10.2005, p. 284-300.

Research output: Contribution to journalArticle

Luker, Kathryn E. ; Hutchens, Martha ; Schultz, Tracey ; Pekosz, Andrew ; Luker, Gary D. / Bioluminescence imaging of vaccinia virus : Effects of interferon on viral replication and spread. In: Virology. 2005 ; Vol. 341, No. 2. pp. 284-300.
@article{a9a0ddc4eb8848ce939bfbe8c9a3641c,
title = "Bioluminescence imaging of vaccinia virus: Effects of interferon on viral replication and spread",
abstract = "Whole animal imaging allows viral replication and localization to be monitored in intact animals, which provides significant advantages for determining viral and host factors that determine pathogenesis. To investigate effects of interferons on spatial and temporal progression of vaccinia infection, we generated recombinant viruses that express firefly luciferase or a monomeric orange fluorescent protein. These viruses allow vaccinia infection to be monitored with bioluminescence or fluorescence imaging, respectively. The recombinant viruses were not attenuated in vitro or in vivo relative to a control WR virus. In cell culture, reporters could be detected readily by 4 h post-infection, showing that these viruses can be used as early markers of infection. The magnitude of firefly luciferase activity measured with bioluminescence imaging in vitro and in vivo correlated directly with increasing titers of vaccinia virus, validating imaging data as a marker of viral infection. Replication of vaccinia was significantly greater in mice lacking receptors for type I interferons (IFN I R-/-) compared with wild-type mice, although both genotypes of mice developed focal infections in lungs and brain after intranasal inoculation. IFN I R-/- mice had greater dissemination of virus to liver and spleen than wild-type animals even when mortality occurred at the same time point after infection. Protective effects of type I interferons were mediated primarily through parenchymal cells rather than hematopoietic cells as analyzed by bone marrow transplant experiments. Collectively, our data define a new function for type I interferon signaling in systemic dissemination of vaccinia and validate these reporter viruses for studies of pathogenesis.",
keywords = "Bioluminescence imaging, Interferon, Vaccinia",
author = "Luker, {Kathryn E.} and Martha Hutchens and Tracey Schultz and Andrew Pekosz and Luker, {Gary D.}",
year = "2005",
month = "10",
day = "25",
doi = "10.1016/j.virol.2005.06.049",
language = "English (US)",
volume = "341",
pages = "284--300",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Bioluminescence imaging of vaccinia virus

T2 - Effects of interferon on viral replication and spread

AU - Luker, Kathryn E.

AU - Hutchens, Martha

AU - Schultz, Tracey

AU - Pekosz, Andrew

AU - Luker, Gary D.

PY - 2005/10/25

Y1 - 2005/10/25

N2 - Whole animal imaging allows viral replication and localization to be monitored in intact animals, which provides significant advantages for determining viral and host factors that determine pathogenesis. To investigate effects of interferons on spatial and temporal progression of vaccinia infection, we generated recombinant viruses that express firefly luciferase or a monomeric orange fluorescent protein. These viruses allow vaccinia infection to be monitored with bioluminescence or fluorescence imaging, respectively. The recombinant viruses were not attenuated in vitro or in vivo relative to a control WR virus. In cell culture, reporters could be detected readily by 4 h post-infection, showing that these viruses can be used as early markers of infection. The magnitude of firefly luciferase activity measured with bioluminescence imaging in vitro and in vivo correlated directly with increasing titers of vaccinia virus, validating imaging data as a marker of viral infection. Replication of vaccinia was significantly greater in mice lacking receptors for type I interferons (IFN I R-/-) compared with wild-type mice, although both genotypes of mice developed focal infections in lungs and brain after intranasal inoculation. IFN I R-/- mice had greater dissemination of virus to liver and spleen than wild-type animals even when mortality occurred at the same time point after infection. Protective effects of type I interferons were mediated primarily through parenchymal cells rather than hematopoietic cells as analyzed by bone marrow transplant experiments. Collectively, our data define a new function for type I interferon signaling in systemic dissemination of vaccinia and validate these reporter viruses for studies of pathogenesis.

AB - Whole animal imaging allows viral replication and localization to be monitored in intact animals, which provides significant advantages for determining viral and host factors that determine pathogenesis. To investigate effects of interferons on spatial and temporal progression of vaccinia infection, we generated recombinant viruses that express firefly luciferase or a monomeric orange fluorescent protein. These viruses allow vaccinia infection to be monitored with bioluminescence or fluorescence imaging, respectively. The recombinant viruses were not attenuated in vitro or in vivo relative to a control WR virus. In cell culture, reporters could be detected readily by 4 h post-infection, showing that these viruses can be used as early markers of infection. The magnitude of firefly luciferase activity measured with bioluminescence imaging in vitro and in vivo correlated directly with increasing titers of vaccinia virus, validating imaging data as a marker of viral infection. Replication of vaccinia was significantly greater in mice lacking receptors for type I interferons (IFN I R-/-) compared with wild-type mice, although both genotypes of mice developed focal infections in lungs and brain after intranasal inoculation. IFN I R-/- mice had greater dissemination of virus to liver and spleen than wild-type animals even when mortality occurred at the same time point after infection. Protective effects of type I interferons were mediated primarily through parenchymal cells rather than hematopoietic cells as analyzed by bone marrow transplant experiments. Collectively, our data define a new function for type I interferon signaling in systemic dissemination of vaccinia and validate these reporter viruses for studies of pathogenesis.

KW - Bioluminescence imaging

KW - Interferon

KW - Vaccinia

UR - http://www.scopus.com/inward/record.url?scp=26444543367&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=26444543367&partnerID=8YFLogxK

U2 - 10.1016/j.virol.2005.06.049

DO - 10.1016/j.virol.2005.06.049

M3 - Article

VL - 341

SP - 284

EP - 300

JO - Virology

JF - Virology

SN - 0042-6822

IS - 2

ER -