Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control

ABSTRACT Ixodes scapularis ticks transmit many infectious agents that cause disease, including tick-borne flaviviruses (TBFVs). TBFV infections cause thousands of human encephalitis cases worldwide annually. In the United States, human TBFV infections with Powassan virus (POWV) are increasing and ha...

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Autores principales:	Jeffrey M. Grabowski, Konstantin A. Tsetsarkin, Dan Long, Dana P. Scott, Rebecca Rosenke, Tom G. Schwan, Luwanika Mlera, Danielle K. Offerdahl, Alexander G. Pletnev, Marshall E. Bloom
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Langat virus Powassan virus RNA interference dsRNA metabolism midgut Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/38fb52528dc24fc5accb0d8e4d1bc6ea
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spelling	oai:doaj.org-article:38fb52528dc24fc5accb0d8e4d1bc6ea2021-11-15T15:51:44ZFlavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control10.1128/mBio.01255-172150-7511https://doaj.org/article/38fb52528dc24fc5accb0d8e4d1bc6ea2017-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01255-17https://doaj.org/toc/2150-7511ABSTRACT Ixodes scapularis ticks transmit many infectious agents that cause disease, including tick-borne flaviviruses (TBFVs). TBFV infections cause thousands of human encephalitis cases worldwide annually. In the United States, human TBFV infections with Powassan virus (POWV) are increasing and have a fatality rate of 10 to 30%. Additionally, Langat virus (LGTV) is a TBFV of low neurovirulence and is used as a model TBFV. TBFV replication and dissemination within I. scapularis organs are poorly characterized, and a deeper understanding of virus biology in this vector may inform effective countermeasures to reduce TBFV transmission. Here, we describe short-term, I. scapularis organ culture models of TBFV infection. Ex vivo organs were metabolically active for 9 to 10 days and were permissive to LGTV and POWV replication. Imaging and videography demonstrated replication and spread of green fluorescent protein-expressing LGTV in the organs. Immunohistochemical staining confirmed LGTV envelope and POWV protein synthesis within the infected organs. LGTV- and POWV-infected organs produced infectious LGTV and POWV; thus, the ex vivo cultures were suitable for study of virus replication in individual organs. LGTV- and POWV-infected midgut and salivary glands were subjected to double-stranded RNA (dsRNA) transfection with dsRNA to the LGTV 3′ untranslated region (UTR), which reduced infectious LGTV and POWV replication, providing a proof-of-concept use of RNA interference in I. scapularis organ cultures to study the effects on TBFV replication. The results contribute important information on TBFV localization within ex vivo I. scapularis organs and provide a significant translational tool for evaluating recombinant, live vaccine candidates and potential tick transcripts and proteins for possible therapeutic use and vaccine development to reduce TBFV transmission. IMPORTANCE Tick-borne flavivirus (TBFV) infections cause neurological and/or hemorrhagic disease in humans worldwide. There are currently no licensed therapeutics or vaccines against Powassan virus (POWV), the only TBFV known to circulate in North America. Evaluating tick vector targets for antitick vaccines directed at reducing TBFV infection within the arthropod vector is a critical step in identifying efficient approaches to controlling TBFV transmission. This study characterized infection of female Ixodes scapularis tick organ cultures of midgut, salivary glands, and synganglion with the low-neurovirulence Langat virus (LGTV) and the more pathogenic POWV. Cell types of specific organs were susceptible to TBFV infection, and a difference in LGTV and POWV replication was noted in TBFV-infected organs. This tick organ culture model of TBFV infection will be useful for various applications, such as screening of tick endogenous dsRNA corresponding to potential control targets within midgut and salivary glands to confirm restriction of TBFV infection.Jeffrey M. GrabowskiKonstantin A. TsetsarkinDan LongDana P. ScottRebecca RosenkeTom G. SchwanLuwanika MleraDanielle K. OfferdahlAlexander G. PletnevMarshall E. BloomAmerican Society for MicrobiologyarticleLangat virusPowassan virusRNA interferencedsRNAmetabolismmidgutMicrobiologyQR1-502ENmBio, Vol 8, Iss 4 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Langat virus Powassan virus RNA interference dsRNA metabolism midgut Microbiology QR1-502
spellingShingle	Langat virus Powassan virus RNA interference dsRNA metabolism midgut Microbiology QR1-502 Jeffrey M. Grabowski Konstantin A. Tsetsarkin Dan Long Dana P. Scott Rebecca Rosenke Tom G. Schwan Luwanika Mlera Danielle K. Offerdahl Alexander G. Pletnev Marshall E. Bloom Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control
description	ABSTRACT Ixodes scapularis ticks transmit many infectious agents that cause disease, including tick-borne flaviviruses (TBFVs). TBFV infections cause thousands of human encephalitis cases worldwide annually. In the United States, human TBFV infections with Powassan virus (POWV) are increasing and have a fatality rate of 10 to 30%. Additionally, Langat virus (LGTV) is a TBFV of low neurovirulence and is used as a model TBFV. TBFV replication and dissemination within I. scapularis organs are poorly characterized, and a deeper understanding of virus biology in this vector may inform effective countermeasures to reduce TBFV transmission. Here, we describe short-term, I. scapularis organ culture models of TBFV infection. Ex vivo organs were metabolically active for 9 to 10 days and were permissive to LGTV and POWV replication. Imaging and videography demonstrated replication and spread of green fluorescent protein-expressing LGTV in the organs. Immunohistochemical staining confirmed LGTV envelope and POWV protein synthesis within the infected organs. LGTV- and POWV-infected organs produced infectious LGTV and POWV; thus, the ex vivo cultures were suitable for study of virus replication in individual organs. LGTV- and POWV-infected midgut and salivary glands were subjected to double-stranded RNA (dsRNA) transfection with dsRNA to the LGTV 3′ untranslated region (UTR), which reduced infectious LGTV and POWV replication, providing a proof-of-concept use of RNA interference in I. scapularis organ cultures to study the effects on TBFV replication. The results contribute important information on TBFV localization within ex vivo I. scapularis organs and provide a significant translational tool for evaluating recombinant, live vaccine candidates and potential tick transcripts and proteins for possible therapeutic use and vaccine development to reduce TBFV transmission. IMPORTANCE Tick-borne flavivirus (TBFV) infections cause neurological and/or hemorrhagic disease in humans worldwide. There are currently no licensed therapeutics or vaccines against Powassan virus (POWV), the only TBFV known to circulate in North America. Evaluating tick vector targets for antitick vaccines directed at reducing TBFV infection within the arthropod vector is a critical step in identifying efficient approaches to controlling TBFV transmission. This study characterized infection of female Ixodes scapularis tick organ cultures of midgut, salivary glands, and synganglion with the low-neurovirulence Langat virus (LGTV) and the more pathogenic POWV. Cell types of specific organs were susceptible to TBFV infection, and a difference in LGTV and POWV replication was noted in TBFV-infected organs. This tick organ culture model of TBFV infection will be useful for various applications, such as screening of tick endogenous dsRNA corresponding to potential control targets within midgut and salivary glands to confirm restriction of TBFV infection.
format	article
author	Jeffrey M. Grabowski Konstantin A. Tsetsarkin Dan Long Dana P. Scott Rebecca Rosenke Tom G. Schwan Luwanika Mlera Danielle K. Offerdahl Alexander G. Pletnev Marshall E. Bloom
author_facet	Jeffrey M. Grabowski Konstantin A. Tsetsarkin Dan Long Dana P. Scott Rebecca Rosenke Tom G. Schwan Luwanika Mlera Danielle K. Offerdahl Alexander G. Pletnev Marshall E. Bloom
author_sort	Jeffrey M. Grabowski
title	Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control
title_short	Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control
title_full	Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control
title_fullStr	Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control
title_full_unstemmed	Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control
title_sort	flavivirus infection of <italic toggle="yes">ixodes scapularis</italic> (black-legged tick) <italic toggle="yes">ex vivo</italic> organotypic cultures and applications for disease control
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/38fb52528dc24fc5accb0d8e4d1bc6ea
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Flavivirus Infection of <italic toggle="yes">Ixodes scapularis</italic> (Black-Legged Tick) <italic toggle="yes">Ex Vivo</italic> Organotypic Cultures and Applications for Disease Control

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