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Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control

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 fata...

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Autores principales: Grabowski, Jeffrey M., Tsetsarkin, Konstantin A., Long, Dan, Scott, Dana P., Rosenke, Rebecca, Schwan, Tom G., Mlera, Luwanika, Offerdahl, Danielle K., Pletnev, Alexander G., Bloom, Marshall E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5565970/
https://www.ncbi.nlm.nih.gov/pubmed/28830948
http://dx.doi.org/10.1128/mBio.01255-17
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author Grabowski, Jeffrey M.
Tsetsarkin, Konstantin A.
Long, Dan
Scott, Dana P.
Rosenke, Rebecca
Schwan, Tom G.
Mlera, Luwanika
Offerdahl, Danielle K.
Pletnev, Alexander G.
Bloom, Marshall E.
author_facet Grabowski, Jeffrey M.
Tsetsarkin, Konstantin A.
Long, Dan
Scott, Dana P.
Rosenke, Rebecca
Schwan, Tom G.
Mlera, Luwanika
Offerdahl, Danielle K.
Pletnev, Alexander G.
Bloom, Marshall E.
author_sort Grabowski, Jeffrey M.
collection PubMed
description 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.
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spelling pubmed-55659702017-08-25 Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control Grabowski, Jeffrey M. Tsetsarkin, Konstantin A. Long, Dan Scott, Dana P. Rosenke, Rebecca Schwan, Tom G. Mlera, Luwanika Offerdahl, Danielle K. Pletnev, Alexander G. Bloom, Marshall E. mBio Research Article 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. American Society for Microbiology 2017-08-22 /pmc/articles/PMC5565970/ /pubmed/28830948 http://dx.doi.org/10.1128/mBio.01255-17 Text en https://www.usa.gov/government-works This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.
spellingShingle Research Article
Grabowski, Jeffrey M.
Tsetsarkin, Konstantin A.
Long, Dan
Scott, Dana P.
Rosenke, Rebecca
Schwan, Tom G.
Mlera, Luwanika
Offerdahl, Danielle K.
Pletnev, Alexander G.
Bloom, Marshall E.
Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control
title Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control
title_full Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control
title_fullStr Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control
title_full_unstemmed Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control
title_short Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control
title_sort flavivirus infection of ixodes scapularis (black-legged tick) ex vivo organotypic cultures and applications for disease control
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5565970/
https://www.ncbi.nlm.nih.gov/pubmed/28830948
http://dx.doi.org/10.1128/mBio.01255-17
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