{"id":{"repo_id":"washington","oai_identifier":"oai:digital.lib.washington.edu:1773/23353"},"canonical_url":"https://search.dev.ndltd.org/etd/washington/oai:digital.lib.washington.edu:1773/23353","repository":{"repo_id":"washington","name":"University of Washington","base_url":"https://digital.lib.washington.edu/server/oai/request"},"display":{"title":"Induction and Regulation of CXCL10 in Hepatocytes During Hepatitis C Virus Infection","abstract":"Chronic Hepatitis C affects an estimated 170 million people worldwide and 4 million in the United States. The pro-inflammatory chemokine CXCL10 is induced by hepatitis C virus (HCV) infection <italic>in vitro</italic> and <italic>in vivo</italic>, and is associated with the outcome of interferon (IFN)-based therapies. Since persistent hepatic inflammation can lead to degenerative liver disease, this work sought to evaluate how innate immune sensors of HCV infection (Toll-like receptor 3 [TLR3] and retinoic acid inducible gene I [RIG-I]) contribute to CXCL10 induction in hepatocytes. CXCL10 mRNA and protein were measured in primary human hepatocytes (PHH) and hepatocyte lines harboring functional or non-functional TLR3 and RIG-I pathways following HCV infection or exposure to receptor-specific stimuli. The contribution of hepatocyte-derived type I and type III IFNs and specific pro-inflammatory transcription factors to CXCL10 induction were also examined. In this study, PHH and immortalized PH5CH8 hepatocytes were confirmed to express functional TLR3 and RIG-I. Specific activation of TLR3 and RIG-I led to CXCL10 induction in a non-synergistic manner, and Huh7 human hepatoma cells expressing both receptors (TLR3+/RIG-I+ Huh7 cells) produced maximal CXCL10 during early HCV infection. Neutralization of type I and type III IFNs had no impact on virus-induced CXCL10 expression in TLR3+/RIG-I+ Huh7 cells, but reduced CXCL10 expression in PHH. PHH cultures were positive for monocyte, macrophage, and dendritic cell mRNAs, suggesting that standard PHH cultures contain non-parenchymal cells (NPCs). Immunodepletion of NPCs eliminated expression of immune and anti- inflammatory markers in PHH cultures, which then showed no IFN requirement for CXCL10 induction during HCV infection. Instead, HCV infection and specific TLR3/RIG-I activation induced binding of NF-κB and IRF3 to the CXCL10 promoter. Together, these data indicate that initial CXCL10 induction in hepatocytes during early HCV infection is independent of hepatocyte-derived type I and type III IFNs, while NPC- and immune cell-derived IFNs contribute to CXCL10 induction during HCV infection in PHH cultures and <italic>in vivo</italic>. Further elucidation of the regulatory pathways controlling CXCL10 induction may reveal novel targets for host-oriented therapies to reduce chronic inflammation, as well as provide insight into the complex and redundant signaling network of the innate immune system.","abstract_html":"Chronic Hepatitis C affects an estimated 170 million people worldwide and 4 million in the United States. The pro-inflammatory chemokine CXCL10 is induced by hepatitis C virus (HCV) infection &lt;italic&gt;in vitro&lt;/italic&gt; and &lt;italic&gt;in vivo&lt;/italic&gt;, and is associated with the outcome of interferon (IFN)-based therapies. Since persistent hepatic inflammation can lead to degenerative liver disease, this work sought to evaluate how innate immune sensors of HCV infection (Toll-like receptor 3 [TLR3] and retinoic acid inducible gene I [RIG-I]) contribute to CXCL10 induction in hepatocytes. CXCL10 mRNA and protein were measured in primary human hepatocytes (PHH) and hepatocyte lines harboring functional or non-functional TLR3 and RIG-I pathways following HCV infection or exposure to receptor-specific stimuli. The contribution of hepatocyte-derived type I and type III IFNs and specific pro-inflammatory transcription factors to CXCL10 induction were also examined. In this study, PHH and immortalized PH5CH8 hepatocytes were confirmed to express functional TLR3 and RIG-I. Specific activation of TLR3 and RIG-I led to CXCL10 induction in a non-synergistic manner, and Huh7 human hepatoma cells expressing both receptors (TLR3+/RIG-I+ Huh7 cells) produced maximal CXCL10 during early HCV infection. Neutralization of type I and type III IFNs had no impact on virus-induced CXCL10 expression in TLR3+/RIG-I+ Huh7 cells, but reduced CXCL10 expression in PHH. PHH cultures were positive for monocyte, macrophage, and dendritic cell mRNAs, suggesting that standard PHH cultures contain non-parenchymal cells (NPCs). Immunodepletion of NPCs eliminated expression of immune and anti- inflammatory markers in PHH cultures, which then showed no IFN requirement for CXCL10 induction during HCV infection. Instead, HCV infection and specific TLR3/RIG-I activation induced binding of NF-κB and IRF3 to the CXCL10 promoter. Together, these data indicate that initial CXCL10 induction in hepatocytes during early HCV infection is independent of hepatocyte-derived type I and type III IFNs, while NPC- and immune cell-derived IFNs contribute to CXCL10 induction during HCV infection in PHH cultures and &lt;italic&gt;in vivo&lt;/italic&gt;. Further elucidation of the regulatory pathways controlling CXCL10 induction may reveal novel targets for host-oriented therapies to reduce chronic inflammation, as well as provide insight into the complex and redundant signaling network of the innate immune system.","abstract_has_math":false,"creators":["Brownell, Jessica Lauren"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Polyak, Stephen J"],"committee_chairs":[],"committee_members":[],"year":2013,"date_issued":"2013-07-25","date_published":"2013-07-25","updated_at":"2026-07-24T05:58:16Z","subjects":["Hepatitis C; Interferon; RIG-I; TLR3"],"languages":["en_US"],"rights":["Copyright is held by the individual authors."],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/1773/23353","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Polyak, Stephen J"]},{"key":"dc:creator","label":"Author","values":["Brownell, Jessica Lauren"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2013-07-25T17:46:41Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2015-12-14T17:55:51Z"]},{"key":"dc:date.issued","label":"Date","values":["2013-07-25"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Hepatitis C; Interferon; RIG-I; TLR3"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en_US"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright is held by the individual authors."]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["Brownell_washington_0250E_11683.pdf"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/1773/23353"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Thesis (Ph.D.)--University of Washington, 2013"]},{"key":"dc:description.abstract","label":"Abstract","values":["Chronic Hepatitis C affects an estimated 170 million people worldwide and 4 million in the United States. The pro-inflammatory chemokine CXCL10 is induced by hepatitis C virus (HCV) infection <italic>in vitro</italic> and <italic>in vivo</italic>, and is associated with the outcome of interferon (IFN)-based therapies. Since persistent hepatic inflammation can lead to degenerative liver disease, this work sought to evaluate how innate immune sensors of HCV infection (Toll-like receptor 3 [TLR3] and retinoic acid inducible gene I [RIG-I]) contribute to CXCL10 induction in hepatocytes. CXCL10 mRNA and protein were measured in primary human hepatocytes (PHH) and hepatocyte lines harboring functional or non-functional TLR3 and RIG-I pathways following HCV infection or exposure to receptor-specific stimuli. The contribution of hepatocyte-derived type I and type III IFNs and specific pro-inflammatory transcription factors to CXCL10 induction were also examined. In this study, PHH and immortalized PH5CH8 hepatocytes were confirmed to express functional TLR3 and RIG-I. Specific activation of TLR3 and RIG-I led to CXCL10 induction in a non-synergistic manner, and Huh7 human hepatoma cells expressing both receptors (TLR3+/RIG-I+ Huh7 cells) produced maximal CXCL10 during early HCV infection. Neutralization of type I and type III IFNs had no impact on virus-induced CXCL10 expression in TLR3+/RIG-I+ Huh7 cells, but reduced CXCL10 expression in PHH. PHH cultures were positive for monocyte, macrophage, and dendritic cell mRNAs, suggesting that standard PHH cultures contain non-parenchymal cells (NPCs). Immunodepletion of NPCs eliminated expression of immune and anti- inflammatory markers in PHH cultures, which then showed no IFN requirement for CXCL10 induction during HCV infection. Instead, HCV infection and specific TLR3/RIG-I activation induced binding of NF-κB and IRF3 to the CXCL10 promoter. Together, these data indicate that initial CXCL10 induction in hepatocytes during early HCV infection is independent of hepatocyte-derived type I and type III IFNs, while NPC- and immune cell-derived IFNs contribute to CXCL10 induction during HCV infection in PHH cultures and <italic>in vivo</italic>. Further elucidation of the regulatory pathways controlling CXCL10 induction may reveal novel targets for host-oriented therapies to reduce chronic inflammation, as well as provide insight into the complex and redundant signaling network of the innate immune system."]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Induction and Regulation of CXCL10 in Hepatocytes During Hepatitis C Virus Infection"]}]}],"canonical_facts":{"dc:contributor.advisor":["Polyak, Stephen J"],"dc:creator":["Brownell, Jessica Lauren"],"dc:date.accessioned":["2013-07-25T17:46:41Z"],"dc:date.available":["2015-12-14T17:55:51Z"],"dc:date.issued":["2013-07-25"],"dc:description":["Thesis (Ph.D.)--University of Washington, 2013"],"dc:description.abstract":["Chronic Hepatitis C affects an estimated 170 million people worldwide and 4 million in the United States. The pro-inflammatory chemokine CXCL10 is induced by hepatitis C virus (HCV) infection <italic>in vitro</italic> and <italic>in vivo</italic>, and is associated with the outcome of interferon (IFN)-based therapies. Since persistent hepatic inflammation can lead to degenerative liver disease, this work sought to evaluate how innate immune sensors of HCV infection (Toll-like receptor 3 [TLR3] and retinoic acid inducible gene I [RIG-I]) contribute to CXCL10 induction in hepatocytes. CXCL10 mRNA and protein were measured in primary human hepatocytes (PHH) and hepatocyte lines harboring functional or non-functional TLR3 and RIG-I pathways following HCV infection or exposure to receptor-specific stimuli. The contribution of hepatocyte-derived type I and type III IFNs and specific pro-inflammatory transcription factors to CXCL10 induction were also examined. In this study, PHH and immortalized PH5CH8 hepatocytes were confirmed to express functional TLR3 and RIG-I. Specific activation of TLR3 and RIG-I led to CXCL10 induction in a non-synergistic manner, and Huh7 human hepatoma cells expressing both receptors (TLR3+/RIG-I+ Huh7 cells) produced maximal CXCL10 during early HCV infection. Neutralization of type I and type III IFNs had no impact on virus-induced CXCL10 expression in TLR3+/RIG-I+ Huh7 cells, but reduced CXCL10 expression in PHH. PHH cultures were positive for monocyte, macrophage, and dendritic cell mRNAs, suggesting that standard PHH cultures contain non-parenchymal cells (NPCs). Immunodepletion of NPCs eliminated expression of immune and anti- inflammatory markers in PHH cultures, which then showed no IFN requirement for CXCL10 induction during HCV infection. Instead, HCV infection and specific TLR3/RIG-I activation induced binding of NF-κB and IRF3 to the CXCL10 promoter. Together, these data indicate that initial CXCL10 induction in hepatocytes during early HCV infection is independent of hepatocyte-derived type I and type III IFNs, while NPC- and immune cell-derived IFNs contribute to CXCL10 induction during HCV infection in PHH cultures and <italic>in vivo</italic>. Further elucidation of the regulatory pathways controlling CXCL10 induction may reveal novel targets for host-oriented therapies to reduce chronic inflammation, as well as provide insight into the complex and redundant signaling network of the innate immune system."],"dc:format.mimetype":["application/pdf"],"dc:identifier.other":["Brownell_washington_0250E_11683.pdf"],"dc:identifier.uri":["http://hdl.handle.net/1773/23353"],"dc:language.iso":["en_US"],"dc:rights":["Copyright is held by the individual authors."],"dc:subject":["Hepatitis C; Interferon; RIG-I; TLR3"],"dc:title":["Induction and Regulation of CXCL10 in Hepatocytes During Hepatitis C Virus Infection"],"dc:type":["Thesis"]},"updated_at":"2026-07-24T05:58:16Z"}