Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivo

Marek's disease virus (MDV) establishes latency in chicken T lymphocytes that can lead to T cell transformation and cancer. Transformed Marek's disease chicken cell lines (MDCCs) can be expanded ex vivo and provide a valuable model to study latency, transformation, and reactivation. Here,...

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Main Authors: Yung-Tien Tien, Haji Akbar, Keith William Jarosinski
Format: Article
Language:English
Published: Frontiers Media S.A. 2023-03-01
Series:Frontiers in Veterinary Science
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fvets.2023.1145757/full
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author Yung-Tien Tien
Haji Akbar
Keith William Jarosinski
author_facet Yung-Tien Tien
Haji Akbar
Keith William Jarosinski
author_sort Yung-Tien Tien
collection DOAJ
description Marek's disease virus (MDV) establishes latency in chicken T lymphocytes that can lead to T cell transformation and cancer. Transformed Marek's disease chicken cell lines (MDCCs) can be expanded ex vivo and provide a valuable model to study latency, transformation, and reactivation. Here, we developed MDCCs from chickens infected with MDV that fluoresce during lytic replication and reactivation. Sodium butyrate treatment increased fluorescent protein expression as evidenced by fluorescent microscopy, flow cytometry, and western blotting; however, it caused significant apoptosis and necrosis. Treatment of MDCCs by decreasing the temperature resulted in robust MDV reactivation without significant induction of apoptosis and necrosis. Furthermore, MDV reactivation was significantly affected by the time in culture that can affect downstream reactivation analyses. In all, our data show that fluorescent protein expression during reactivation is a robust tool to examine viral replication in live cells ex vivo, and temperature treatment is an efficient technique to induce reactivation without punitive effects on cell viability seen with chemical treatment.
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spelling doaj.art-cf69edc2d6284795a16ecbe6a259f4f72023-03-08T05:31:02ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692023-03-011010.3389/fvets.2023.11457571145757Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivoYung-Tien TienHaji AkbarKeith William JarosinskiMarek's disease virus (MDV) establishes latency in chicken T lymphocytes that can lead to T cell transformation and cancer. Transformed Marek's disease chicken cell lines (MDCCs) can be expanded ex vivo and provide a valuable model to study latency, transformation, and reactivation. Here, we developed MDCCs from chickens infected with MDV that fluoresce during lytic replication and reactivation. Sodium butyrate treatment increased fluorescent protein expression as evidenced by fluorescent microscopy, flow cytometry, and western blotting; however, it caused significant apoptosis and necrosis. Treatment of MDCCs by decreasing the temperature resulted in robust MDV reactivation without significant induction of apoptosis and necrosis. Furthermore, MDV reactivation was significantly affected by the time in culture that can affect downstream reactivation analyses. In all, our data show that fluorescent protein expression during reactivation is a robust tool to examine viral replication in live cells ex vivo, and temperature treatment is an efficient technique to induce reactivation without punitive effects on cell viability seen with chemical treatment.https://www.frontiersin.org/articles/10.3389/fvets.2023.1145757/fullMarek's disease (MD)reactivationherpesviruschickentransformation
spellingShingle Yung-Tien Tien
Haji Akbar
Keith William Jarosinski
Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivo
Frontiers in Veterinary Science
Marek's disease (MD)
reactivation
herpesvirus
chicken
transformation
title Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivo
title_full Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivo
title_fullStr Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivo
title_full_unstemmed Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivo
title_short Temperature-induced reactivation of Marek's disease virus-transformed T cells ex vivo
title_sort temperature induced reactivation of marek s disease virus transformed t cells ex vivo
topic Marek's disease (MD)
reactivation
herpesvirus
chicken
transformation
url https://www.frontiersin.org/articles/10.3389/fvets.2023.1145757/full
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