A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection
Hepatitis B virus (HBV) infection, which increases the risk of cirrhosis and hepatocellular carcinoma and requires lifelong treatment, has become a major global health problem. However, host factors essential to the HBV life cycle are still unclear, and the development of new drugs is needed. Cells...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2019-07, Vol.515 (1), p.156-162 |
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| creator | Ueda, Youki Gu, Weilin Dansako, Hiromichi Nishitsuji, Hironori Satoh, Shinya Shimotohno, Kunitada Kato, Nobuyuki |
| description | Hepatitis B virus (HBV) infection, which increases the risk of cirrhosis and hepatocellular carcinoma and requires lifelong treatment, has become a major global health problem. However, host factors essential to the HBV life cycle are still unclear, and the development of new drugs is needed.
Cells derived from the human hepatoma cell line HepG2 and engineered to overexpress sodium taurocholate cotransporting polypeptide (NTCP: a receptor for HBV), termed HepG2/NTCP cells, are widely used as the cell-based HBV infection and replication systems for HBV research. We recently found that human hepatoma cell line Li23-derived cells overexpressing NTCP (A8 cells subcloned from Li23 cells), whose gene expression profile was distinct from that of HepG2/NTCP cells, were also sensitive to HBV infection. However, the HBV susceptibility of A8 cells was around 1/100 that of HepG2/NTCP cells. Since we considered that plural cell assay systems will be needed for the objective evaluation of anti-HBV reagents, as we previously demonstrated in hepatitis C virus research, we here attempted to develop a new Li23 cell-derived assay system equivalent to that using HepG2/NTCP cells. By repeated subcloning of A8 cells, we successfully established a new cell line (A8.15.78.10) exhibiting high HBV susceptibility equal to that of HepG2/NTCP cells. Characterization of A8.15.78.10 cells revealed that the increase of HBV susceptibility was correlated with increases in the protein and glycosylation levels of NTCP, and with decreased expression of STING, a factor contributing to innate immunity. Finally, we performed a comparative evaluation of HBV entry inhibitors (cyclosporin A and rosiglitazone) by an HBV/secNL reporter assay using A8.15.78.10 cells or HepG2/NTCP cells. The results confirmed that cyclosporin A exhibited anti-HBV activity in both cell lines, as previously reported. However, we found that rosiglitazone did not show the anti-HBV activity in A8.15.78.10 cells, although it worked in HepG2/NTCP cells as previously reported. This suggested that the difference in anti-HBV activity between cyclosporin A and rosiglitazone was due to the different types of cells used for the assay. In conclusion, plural assay systems using different types of cells are required for the objective and impartial evaluation of anti-HBV reagents.
•We established a Li23/NTCP subcloned cell line with high HBV susceptibility.•The glycosylation level of NTCP may be involved in HBV susceptibility.•A low le |
| doi_str_mv | 10.1016/j.bbrc.2019.05.126 |
| format | Article |
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Cells derived from the human hepatoma cell line HepG2 and engineered to overexpress sodium taurocholate cotransporting polypeptide (NTCP: a receptor for HBV), termed HepG2/NTCP cells, are widely used as the cell-based HBV infection and replication systems for HBV research. We recently found that human hepatoma cell line Li23-derived cells overexpressing NTCP (A8 cells subcloned from Li23 cells), whose gene expression profile was distinct from that of HepG2/NTCP cells, were also sensitive to HBV infection. However, the HBV susceptibility of A8 cells was around 1/100 that of HepG2/NTCP cells. Since we considered that plural cell assay systems will be needed for the objective evaluation of anti-HBV reagents, as we previously demonstrated in hepatitis C virus research, we here attempted to develop a new Li23 cell-derived assay system equivalent to that using HepG2/NTCP cells. By repeated subcloning of A8 cells, we successfully established a new cell line (A8.15.78.10) exhibiting high HBV susceptibility equal to that of HepG2/NTCP cells. Characterization of A8.15.78.10 cells revealed that the increase of HBV susceptibility was correlated with increases in the protein and glycosylation levels of NTCP, and with decreased expression of STING, a factor contributing to innate immunity. Finally, we performed a comparative evaluation of HBV entry inhibitors (cyclosporin A and rosiglitazone) by an HBV/secNL reporter assay using A8.15.78.10 cells or HepG2/NTCP cells. The results confirmed that cyclosporin A exhibited anti-HBV activity in both cell lines, as previously reported. However, we found that rosiglitazone did not show the anti-HBV activity in A8.15.78.10 cells, although it worked in HepG2/NTCP cells as previously reported. This suggested that the difference in anti-HBV activity between cyclosporin A and rosiglitazone was due to the different types of cells used for the assay. In conclusion, plural assay systems using different types of cells are required for the objective and impartial evaluation of anti-HBV reagents.
•We established a Li23/NTCP subcloned cell line with high HBV susceptibility.•The glycosylation level of NTCP may be involved in HBV susceptibility.•A low level of STING expression may be involved in HBV susceptibility.•The established cell line was useful for objective evaluation of anti-HBV reagents.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2019.05.126</identifier><identifier>PMID: 31133379</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Hepatitis B virus ; Li23/NTCP ; NTCP glycosylation ; Plural assay systems ; STING ; Subcloning</subject><ispartof>Biochemical and biophysical research communications, 2019-07, Vol.515 (1), p.156-162</ispartof><rights>2019 Elsevier Inc.</rights><rights>Copyright © 2019 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c422t-9472bd423affa5aee6cfa70462ade30495ffc773a5d16a0918c74894ea2676a53</citedby><cites>FETCH-LOGICAL-c422t-9472bd423affa5aee6cfa70462ade30495ffc773a5d16a0918c74894ea2676a53</cites><orcidid>0000-0002-2627-8524 ; 0000-0002-2353-3024</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbrc.2019.05.126$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,782,786,3554,27933,27934,46004</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31133379$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ueda, Youki</creatorcontrib><creatorcontrib>Gu, Weilin</creatorcontrib><creatorcontrib>Dansako, Hiromichi</creatorcontrib><creatorcontrib>Nishitsuji, Hironori</creatorcontrib><creatorcontrib>Satoh, Shinya</creatorcontrib><creatorcontrib>Shimotohno, Kunitada</creatorcontrib><creatorcontrib>Kato, Nobuyuki</creatorcontrib><title>A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>Hepatitis B virus (HBV) infection, which increases the risk of cirrhosis and hepatocellular carcinoma and requires lifelong treatment, has become a major global health problem. However, host factors essential to the HBV life cycle are still unclear, and the development of new drugs is needed.
Cells derived from the human hepatoma cell line HepG2 and engineered to overexpress sodium taurocholate cotransporting polypeptide (NTCP: a receptor for HBV), termed HepG2/NTCP cells, are widely used as the cell-based HBV infection and replication systems for HBV research. We recently found that human hepatoma cell line Li23-derived cells overexpressing NTCP (A8 cells subcloned from Li23 cells), whose gene expression profile was distinct from that of HepG2/NTCP cells, were also sensitive to HBV infection. However, the HBV susceptibility of A8 cells was around 1/100 that of HepG2/NTCP cells. Since we considered that plural cell assay systems will be needed for the objective evaluation of anti-HBV reagents, as we previously demonstrated in hepatitis C virus research, we here attempted to develop a new Li23 cell-derived assay system equivalent to that using HepG2/NTCP cells. By repeated subcloning of A8 cells, we successfully established a new cell line (A8.15.78.10) exhibiting high HBV susceptibility equal to that of HepG2/NTCP cells. Characterization of A8.15.78.10 cells revealed that the increase of HBV susceptibility was correlated with increases in the protein and glycosylation levels of NTCP, and with decreased expression of STING, a factor contributing to innate immunity. Finally, we performed a comparative evaluation of HBV entry inhibitors (cyclosporin A and rosiglitazone) by an HBV/secNL reporter assay using A8.15.78.10 cells or HepG2/NTCP cells. The results confirmed that cyclosporin A exhibited anti-HBV activity in both cell lines, as previously reported. However, we found that rosiglitazone did not show the anti-HBV activity in A8.15.78.10 cells, although it worked in HepG2/NTCP cells as previously reported. This suggested that the difference in anti-HBV activity between cyclosporin A and rosiglitazone was due to the different types of cells used for the assay. In conclusion, plural assay systems using different types of cells are required for the objective and impartial evaluation of anti-HBV reagents.
•We established a Li23/NTCP subcloned cell line with high HBV susceptibility.•The glycosylation level of NTCP may be involved in HBV susceptibility.•A low level of STING expression may be involved in HBV susceptibility.•The established cell line was useful for objective evaluation of anti-HBV reagents.</description><subject>Hepatitis B virus</subject><subject>Li23/NTCP</subject><subject>NTCP glycosylation</subject><subject>Plural assay systems</subject><subject>STING</subject><subject>Subcloning</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNp9kEtLw0AUhQdRbH38ARcySzeJdx6ZdMBNLb6g4EbF3TCZ3NgpaVJnEh__3pRWl64uXL5z4HyEnDFIGTB1uUyLIriUA9MpZCnjao-MGWhIOAO5T8YAoBKu2euIHMW4BGBMKn1IRoIxIUSux-RlShv8pAtc265dWeqwrmntG6T4tfCF73zzRhf-bUFjHx2uu-FX--6bdu02NBCRXtMPH_pIfVOh63zbnJCDytYRT3f3mDzf3jzN7pP5493DbDpPnOS8S7TMeVFKLmxV2cwiKlfZHKTitkQBUmdV5fJc2KxkyoJmE5fLiZZoucqVzcQxudj2rkP73mPszMrHzQbbYNtHw7ngoCd8wgeUb1EX2hgDVmYd_MqGb8PAbHyapdn4NBufBjIz-BxC57v-vlhh-Rf5FTgAV1sAh5UfHoOJzmPjsPRhUGHK1v_X_wN4xYci</recordid><startdate>20190712</startdate><enddate>20190712</enddate><creator>Ueda, Youki</creator><creator>Gu, Weilin</creator><creator>Dansako, Hiromichi</creator><creator>Nishitsuji, Hironori</creator><creator>Satoh, Shinya</creator><creator>Shimotohno, Kunitada</creator><creator>Kato, Nobuyuki</creator><general>Elsevier Inc</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-2627-8524</orcidid><orcidid>https://orcid.org/0000-0002-2353-3024</orcidid></search><sort><creationdate>20190712</creationdate><title>A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection</title><author>Ueda, Youki ; Gu, Weilin ; Dansako, Hiromichi ; Nishitsuji, Hironori ; Satoh, Shinya ; Shimotohno, Kunitada ; Kato, Nobuyuki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c422t-9472bd423affa5aee6cfa70462ade30495ffc773a5d16a0918c74894ea2676a53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Hepatitis B virus</topic><topic>Li23/NTCP</topic><topic>NTCP glycosylation</topic><topic>Plural assay systems</topic><topic>STING</topic><topic>Subcloning</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ueda, Youki</creatorcontrib><creatorcontrib>Gu, Weilin</creatorcontrib><creatorcontrib>Dansako, Hiromichi</creatorcontrib><creatorcontrib>Nishitsuji, Hironori</creatorcontrib><creatorcontrib>Satoh, Shinya</creatorcontrib><creatorcontrib>Shimotohno, Kunitada</creatorcontrib><creatorcontrib>Kato, Nobuyuki</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ueda, Youki</au><au>Gu, Weilin</au><au>Dansako, Hiromichi</au><au>Nishitsuji, Hironori</au><au>Satoh, Shinya</au><au>Shimotohno, Kunitada</au><au>Kato, Nobuyuki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2019-07-12</date><risdate>2019</risdate><volume>515</volume><issue>1</issue><spage>156</spage><epage>162</epage><pages>156-162</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>Hepatitis B virus (HBV) infection, which increases the risk of cirrhosis and hepatocellular carcinoma and requires lifelong treatment, has become a major global health problem. However, host factors essential to the HBV life cycle are still unclear, and the development of new drugs is needed.
Cells derived from the human hepatoma cell line HepG2 and engineered to overexpress sodium taurocholate cotransporting polypeptide (NTCP: a receptor for HBV), termed HepG2/NTCP cells, are widely used as the cell-based HBV infection and replication systems for HBV research. We recently found that human hepatoma cell line Li23-derived cells overexpressing NTCP (A8 cells subcloned from Li23 cells), whose gene expression profile was distinct from that of HepG2/NTCP cells, were also sensitive to HBV infection. However, the HBV susceptibility of A8 cells was around 1/100 that of HepG2/NTCP cells. Since we considered that plural cell assay systems will be needed for the objective evaluation of anti-HBV reagents, as we previously demonstrated in hepatitis C virus research, we here attempted to develop a new Li23 cell-derived assay system equivalent to that using HepG2/NTCP cells. By repeated subcloning of A8 cells, we successfully established a new cell line (A8.15.78.10) exhibiting high HBV susceptibility equal to that of HepG2/NTCP cells. Characterization of A8.15.78.10 cells revealed that the increase of HBV susceptibility was correlated with increases in the protein and glycosylation levels of NTCP, and with decreased expression of STING, a factor contributing to innate immunity. Finally, we performed a comparative evaluation of HBV entry inhibitors (cyclosporin A and rosiglitazone) by an HBV/secNL reporter assay using A8.15.78.10 cells or HepG2/NTCP cells. The results confirmed that cyclosporin A exhibited anti-HBV activity in both cell lines, as previously reported. However, we found that rosiglitazone did not show the anti-HBV activity in A8.15.78.10 cells, although it worked in HepG2/NTCP cells as previously reported. This suggested that the difference in anti-HBV activity between cyclosporin A and rosiglitazone was due to the different types of cells used for the assay. In conclusion, plural assay systems using different types of cells are required for the objective and impartial evaluation of anti-HBV reagents.
•We established a Li23/NTCP subcloned cell line with high HBV susceptibility.•The glycosylation level of NTCP may be involved in HBV susceptibility.•A low level of STING expression may be involved in HBV susceptibility.•The established cell line was useful for objective evaluation of anti-HBV reagents.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>31133379</pmid><doi>10.1016/j.bbrc.2019.05.126</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0002-2627-8524</orcidid><orcidid>https://orcid.org/0000-0002-2353-3024</orcidid></addata></record> |
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| subjects | Hepatitis B virus Li23/NTCP NTCP glycosylation Plural assay systems STING Subcloning |
| title | A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection |
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