An Ixodes scapularis glutathione S-transferase plays a role in cell survival and viability during Langat virus infection of a tick cell line
•Increased IsGST1 gene expression was observed on day 2 of LGTV infection.•IsGST1 gene knockdown results in decreased ISE6 cell viability.•IsGST1 gene knockdown and LGTV infection caused increased ISE6 cell mortality.•IsGST1 gene knockdown decreases LGTV proliferation in ISE6 cells. Ticks are import...
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description | •Increased IsGST1 gene expression was observed on day 2 of LGTV infection.•IsGST1 gene knockdown results in decreased ISE6 cell viability.•IsGST1 gene knockdown and LGTV infection caused increased ISE6 cell mortality.•IsGST1 gene knockdown decreases LGTV proliferation in ISE6 cells.
Ticks are important vectors of diseases affecting both humans and animals. To be an efficient vector, ticks have to survive infection by pathogens such as the Langat virus (LGTV). One method utilized by ticks is their complex antioxidant mechanism. Included in the vast antioxidant processes are several enzymes involved in redox homeostasis. The ubiquitous glutathione S-transferases (GSTs) belong to the antioxidant family of enzymes. In this study, we evaluated the role of a GST during LGTV infection.
ISE6 cells were infected with LGTV with a multiplicity of infection (MOI) of 0.01 and observed daily. The infection success was monitored via indirect immunofluorescent antibody test (IFAT) for LGTV for up to 4 days. The gene expression of IsGST1 was determined by real-time polymerase chain reaction (PCR) using IsGST1 gene–specific primers. Knockdown of the IsGST1 gene with subsequent LGTV infection was also performed. Afterward, ISE6 cell mortality and viability were checked daily until the fourth day. The virus titer from supernatants of IsGST1-knockdown cells was quantified using a focus-formation assay.
IFAT data showed that LGTV infects ISE6 cells in a time-dependent manner with increasing infection from day 0 to day 4. The IsGST1 genes showed an increasing expression until day 2 of infection, while decreased expression was observed from day 3 to day 4 post-infection. Knockdown of the IsGST1 resulted in increased mortality on the third day of infection, while the cell viability was also negatively affected by the knockdown of the IsGST1 genes from day 0 to day 4 post-infection. Knockdown of the IsGST1 genes also resulted in a decreased viral titer from the supernatants of the ISE6 cells infected with LGTV.
Based on the results, GSTs are possibly utilized both by cells and the virus for mutual survival and proliferation.
[Display omitted] |
doi_str_mv | 10.1016/j.actatropica.2020.105763 |
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Ticks are important vectors of diseases affecting both humans and animals. To be an efficient vector, ticks have to survive infection by pathogens such as the Langat virus (LGTV). One method utilized by ticks is their complex antioxidant mechanism. Included in the vast antioxidant processes are several enzymes involved in redox homeostasis. The ubiquitous glutathione S-transferases (GSTs) belong to the antioxidant family of enzymes. In this study, we evaluated the role of a GST during LGTV infection.
ISE6 cells were infected with LGTV with a multiplicity of infection (MOI) of 0.01 and observed daily. The infection success was monitored via indirect immunofluorescent antibody test (IFAT) for LGTV for up to 4 days. The gene expression of IsGST1 was determined by real-time polymerase chain reaction (PCR) using IsGST1 gene–specific primers. Knockdown of the IsGST1 gene with subsequent LGTV infection was also performed. Afterward, ISE6 cell mortality and viability were checked daily until the fourth day. The virus titer from supernatants of IsGST1-knockdown cells was quantified using a focus-formation assay.
IFAT data showed that LGTV infects ISE6 cells in a time-dependent manner with increasing infection from day 0 to day 4. The IsGST1 genes showed an increasing expression until day 2 of infection, while decreased expression was observed from day 3 to day 4 post-infection. Knockdown of the IsGST1 resulted in increased mortality on the third day of infection, while the cell viability was also negatively affected by the knockdown of the IsGST1 genes from day 0 to day 4 post-infection. Knockdown of the IsGST1 genes also resulted in a decreased viral titer from the supernatants of the ISE6 cells infected with LGTV.
Based on the results, GSTs are possibly utilized both by cells and the virus for mutual survival and proliferation.
[Display omitted]</description><identifier>ISSN: 0001-706X</identifier><identifier>EISSN: 1873-6254</identifier><identifier>DOI: 10.1016/j.actatropica.2020.105763</identifier><identifier>PMID: 33242485</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Cell Line ; Cell Survival - physiology ; Encephalitis Viruses, Tick-Borne - genetics ; Encephalitis Viruses, Tick-Borne - physiology ; Gene Expression Regulation, Enzymologic ; Gene Knockdown Techniques ; Glutathione Transferase - metabolism ; GST ; ISE6 cells ; Ixodes - cytology ; Ixodes - enzymology ; Langat virus ; Real-Time Polymerase Chain Reaction ; tick</subject><ispartof>Acta tropica, 2021-02, Vol.214, p.105763-105763, Article 105763</ispartof><rights>2020 Elsevier B.V.</rights><rights>Copyright © 2020 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c377t-fa83a35d32baec8bd5e858bb4bf9af0ca0eeb03a45e88a6654be813b5690887d3</citedby><cites>FETCH-LOGICAL-c377t-fa83a35d32baec8bd5e858bb4bf9af0ca0eeb03a45e88a6654be813b5690887d3</cites><orcidid>0000-0002-3141-5159</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.actatropica.2020.105763$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33242485$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hernandez, Emmanuel Pacia</creatorcontrib><creatorcontrib>Talactac, Melbourne Rio</creatorcontrib><creatorcontrib>Vitor, Rodel Jonathan Santos</creatorcontrib><creatorcontrib>Yoshii, Kentaro</creatorcontrib><creatorcontrib>Tanaka, Tetsuya</creatorcontrib><title>An Ixodes scapularis glutathione S-transferase plays a role in cell survival and viability during Langat virus infection of a tick cell line</title><title>Acta tropica</title><addtitle>Acta Trop</addtitle><description>•Increased IsGST1 gene expression was observed on day 2 of LGTV infection.•IsGST1 gene knockdown results in decreased ISE6 cell viability.•IsGST1 gene knockdown and LGTV infection caused increased ISE6 cell mortality.•IsGST1 gene knockdown decreases LGTV proliferation in ISE6 cells.
Ticks are important vectors of diseases affecting both humans and animals. To be an efficient vector, ticks have to survive infection by pathogens such as the Langat virus (LGTV). One method utilized by ticks is their complex antioxidant mechanism. Included in the vast antioxidant processes are several enzymes involved in redox homeostasis. The ubiquitous glutathione S-transferases (GSTs) belong to the antioxidant family of enzymes. In this study, we evaluated the role of a GST during LGTV infection.
ISE6 cells were infected with LGTV with a multiplicity of infection (MOI) of 0.01 and observed daily. The infection success was monitored via indirect immunofluorescent antibody test (IFAT) for LGTV for up to 4 days. The gene expression of IsGST1 was determined by real-time polymerase chain reaction (PCR) using IsGST1 gene–specific primers. Knockdown of the IsGST1 gene with subsequent LGTV infection was also performed. Afterward, ISE6 cell mortality and viability were checked daily until the fourth day. The virus titer from supernatants of IsGST1-knockdown cells was quantified using a focus-formation assay.
IFAT data showed that LGTV infects ISE6 cells in a time-dependent manner with increasing infection from day 0 to day 4. The IsGST1 genes showed an increasing expression until day 2 of infection, while decreased expression was observed from day 3 to day 4 post-infection. Knockdown of the IsGST1 resulted in increased mortality on the third day of infection, while the cell viability was also negatively affected by the knockdown of the IsGST1 genes from day 0 to day 4 post-infection. Knockdown of the IsGST1 genes also resulted in a decreased viral titer from the supernatants of the ISE6 cells infected with LGTV.
Based on the results, GSTs are possibly utilized both by cells and the virus for mutual survival and proliferation.
[Display omitted]</description><subject>Animals</subject><subject>Cell Line</subject><subject>Cell Survival - physiology</subject><subject>Encephalitis Viruses, Tick-Borne - genetics</subject><subject>Encephalitis Viruses, Tick-Borne - physiology</subject><subject>Gene Expression Regulation, Enzymologic</subject><subject>Gene Knockdown Techniques</subject><subject>Glutathione Transferase - metabolism</subject><subject>GST</subject><subject>ISE6 cells</subject><subject>Ixodes - cytology</subject><subject>Ixodes - enzymology</subject><subject>Langat virus</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>tick</subject><issn>0001-706X</issn><issn>1873-6254</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc2O0zAUhS3EiCkDr4DMjk2KY8eJuxxV_IxUiQUz0uysa-emuLhOsJ2KvgMPjasMiCUry0fnfNe-h5C3NVvXrG7fH9ZgM-Q4Ts7CmjN-0WXXimdkVatOVC2XzXOyYozVVcfax2vyMqVDufFO8hfkWgje8EbJFfl1G-jdz7HHRJOFafYQXaJ7Pxf-NzcGpF-rHCGkASMkpJOHc6JA4-iRukAtek_THE_uBJ5C6OnJgXHe5TPt5-jCnu4g7CEXPc6pRAa0uYDpOBRMdvb7wvAu4CtyNYBP-PrpvCEPHz_cbz9Xuy-f7ra3u8qKrsvVAEqAkL3gBtAq00tUUhnTmGEDA7PAEA0T0BRdQdvKxqCqhZHthinV9eKGvFu4Uxx_zJiyPrp0eQUEHOekeVMyTV1LXqybxWrjmFLEQU_RHSGedc30pQx90P-UoS9l6KWMkn3zNGY2R-z_Jv9svxi2iwHLZ08Oo07WYbDYu1i2pPvR_ceY34M-pEY</recordid><startdate>202102</startdate><enddate>202102</enddate><creator>Hernandez, Emmanuel Pacia</creator><creator>Talactac, Melbourne Rio</creator><creator>Vitor, Rodel Jonathan Santos</creator><creator>Yoshii, Kentaro</creator><creator>Tanaka, Tetsuya</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-3141-5159</orcidid></search><sort><creationdate>202102</creationdate><title>An Ixodes scapularis glutathione S-transferase plays a role in cell survival and viability during Langat virus infection of a tick cell line</title><author>Hernandez, Emmanuel Pacia ; Talactac, Melbourne Rio ; Vitor, Rodel Jonathan Santos ; Yoshii, Kentaro ; Tanaka, Tetsuya</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c377t-fa83a35d32baec8bd5e858bb4bf9af0ca0eeb03a45e88a6654be813b5690887d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Animals</topic><topic>Cell Line</topic><topic>Cell Survival - physiology</topic><topic>Encephalitis Viruses, Tick-Borne - genetics</topic><topic>Encephalitis Viruses, Tick-Borne - physiology</topic><topic>Gene Expression Regulation, Enzymologic</topic><topic>Gene Knockdown Techniques</topic><topic>Glutathione Transferase - metabolism</topic><topic>GST</topic><topic>ISE6 cells</topic><topic>Ixodes - cytology</topic><topic>Ixodes - enzymology</topic><topic>Langat virus</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>tick</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hernandez, Emmanuel Pacia</creatorcontrib><creatorcontrib>Talactac, Melbourne Rio</creatorcontrib><creatorcontrib>Vitor, Rodel Jonathan Santos</creatorcontrib><creatorcontrib>Yoshii, Kentaro</creatorcontrib><creatorcontrib>Tanaka, Tetsuya</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Acta tropica</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hernandez, Emmanuel Pacia</au><au>Talactac, Melbourne Rio</au><au>Vitor, Rodel Jonathan Santos</au><au>Yoshii, Kentaro</au><au>Tanaka, Tetsuya</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An Ixodes scapularis glutathione S-transferase plays a role in cell survival and viability during Langat virus infection of a tick cell line</atitle><jtitle>Acta tropica</jtitle><addtitle>Acta Trop</addtitle><date>2021-02</date><risdate>2021</risdate><volume>214</volume><spage>105763</spage><epage>105763</epage><pages>105763-105763</pages><artnum>105763</artnum><issn>0001-706X</issn><eissn>1873-6254</eissn><abstract>•Increased IsGST1 gene expression was observed on day 2 of LGTV infection.•IsGST1 gene knockdown results in decreased ISE6 cell viability.•IsGST1 gene knockdown and LGTV infection caused increased ISE6 cell mortality.•IsGST1 gene knockdown decreases LGTV proliferation in ISE6 cells.
Ticks are important vectors of diseases affecting both humans and animals. To be an efficient vector, ticks have to survive infection by pathogens such as the Langat virus (LGTV). One method utilized by ticks is their complex antioxidant mechanism. Included in the vast antioxidant processes are several enzymes involved in redox homeostasis. The ubiquitous glutathione S-transferases (GSTs) belong to the antioxidant family of enzymes. In this study, we evaluated the role of a GST during LGTV infection.
ISE6 cells were infected with LGTV with a multiplicity of infection (MOI) of 0.01 and observed daily. The infection success was monitored via indirect immunofluorescent antibody test (IFAT) for LGTV for up to 4 days. The gene expression of IsGST1 was determined by real-time polymerase chain reaction (PCR) using IsGST1 gene–specific primers. Knockdown of the IsGST1 gene with subsequent LGTV infection was also performed. Afterward, ISE6 cell mortality and viability were checked daily until the fourth day. The virus titer from supernatants of IsGST1-knockdown cells was quantified using a focus-formation assay.
IFAT data showed that LGTV infects ISE6 cells in a time-dependent manner with increasing infection from day 0 to day 4. The IsGST1 genes showed an increasing expression until day 2 of infection, while decreased expression was observed from day 3 to day 4 post-infection. Knockdown of the IsGST1 resulted in increased mortality on the third day of infection, while the cell viability was also negatively affected by the knockdown of the IsGST1 genes from day 0 to day 4 post-infection. Knockdown of the IsGST1 genes also resulted in a decreased viral titer from the supernatants of the ISE6 cells infected with LGTV.
Based on the results, GSTs are possibly utilized both by cells and the virus for mutual survival and proliferation.
[Display omitted]</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>33242485</pmid><doi>10.1016/j.actatropica.2020.105763</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0002-3141-5159</orcidid></addata></record> |
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subjects | Animals Cell Line Cell Survival - physiology Encephalitis Viruses, Tick-Borne - genetics Encephalitis Viruses, Tick-Borne - physiology Gene Expression Regulation, Enzymologic Gene Knockdown Techniques Glutathione Transferase - metabolism GST ISE6 cells Ixodes - cytology Ixodes - enzymology Langat virus Real-Time Polymerase Chain Reaction tick |
title | An Ixodes scapularis glutathione S-transferase plays a role in cell survival and viability during Langat virus infection of a tick cell line |
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