Ian N Hines
East Carolina University
Project start date: 2007-08-01
Project end date: 2012-02-29
Sponsored Links Excellgen http://Excellgen.com
INVOLVEMENT OF NATURAL KILLER T CELLS IN HEPATIC FIBROGENESIS
Ian N Hines, Assistant Professor
University Of North Carolina Chapel Hill, Office Of Sponsored Research, Chapel Hill, Nc 27599
Grant 3K01AA016563-03S1 from National Institute On Alcohol Abuse And Alcoholism
Abstract: Hepatic fibrosis is a consequence of a number of chronic liver pathologies including alcoholic liver disease, viral infection, and biliary outflow obstruction. Growing experimental evidence implicates the class of cytokines produced, specifically the T helper (Th) cytokine phenotype expressed, as a key regulator of the fibrotic response. Indeed, while Th1 type cytokines such as interferon gamma (IFNg) or interleukin (IL) 12 are associated with hepatitis and acute inflammation, expression of the Th2 type cytokines IL4,IL5,andIL13 drive fibrosis development. Moreover, it appears that during chronic liver diseases a switch occurs in which a balanced or perhaps Th1-shifted cytokine response is replaced by an unbalanced Th2 predominated cytokine phenotype. Natural killer T (NKT) cells make up approximately 50% of the intrahepatic lymphocytes and are capable of producing both Th1 and Th2 type cytokines making them a potential regulator of the hepatic Th cytokine balance. The impact this cell population has on the hepatic fibrotic response is not known. It is hypothesized that NKT cells limit hepatocellular inflammation and promote hepatic fibrosis through production of key regulatory and pro-fibrotic Th2 type cytokines. Utilizing two well described models of hepatic fibrosis in rodents, carbon tetrachloride administration or bile duct ligation, and two mouse models of NKT cell deficiency, CD1d-/- mice and Ja281-/- mice, the role that NKT cells play in the fibrotic repsonse will be evaulated. In the first aim, the direct consequence of a deficiency in hepatic NKT cells in the development of hepatic fibrogenesis will be determined. Furthermore, using in vitro cell coculture, the role of NKT cells and NKT cell derived factors in the activation primary hepatic stellate cells will also be examined. In the second aim, hepatic NKT cell populations will be exhaustively characterized prior to and during the development of hepatic fibrosis focusing on their Th cytokine phenotype. The third aim will extend these findings and determine the role of NKT cell derived cytokines specifically using a novel adoptive transfer approach where NKT cell deficient mice are reconstituted with certain cytokine deficient NKT cells including IFNg deficient and IL4 deficient cells. The final aim will address the role of IL12, a known inhibitor of NKT cell function and survival, in the development and progression of experimental fibrosis. Together, these studies will provide new insight into the mechanisms of T cell-mediated hepatic fibrosis
Keywords: A Mouse; Acute Hepatitis; Address; Adoptive Transfer; Alcoholic Liver Diseases; Antibodies; B Cell Differentiation Factor I Gene; B-Cell Stimulatory Factor 1 Gene; BSF-1 Gene; BSF1 Gene; Bile Ducts; Bile duct structure; Biliary; Blood Serum; Body Tissues; Carbon Tetrachloride; Cell Count; Cell Fraction; Cell Function; Cell Isolation; Cell Number; Cell Process; Cell Segregation; Cell Separation; Cell Separation Technology; Cell physiology; Cell-Extracellular Matrix; Cell/Tissue, Immunohistochemistry; Cells; Cellular Function; Cellular Physiology; Cellular Process; Chronic; Closure by Ligation; Co-culture; Cocultivation; Coculture; Coculture Techniques; Collagen; Contractile Proteins; Cytofluorometry, Flow; Data; Detection; Development; ECM; EDF Gene; Edodekin Alfa; Eosinophil Differentiation Factor Gene; Equilibrium; Extracellular Matrix; Fibrosis; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Gamma interferon; Hepatic; Hepatic Disorder; Hepatic Fibrogenesis; Hepatic Stellate Cell; IFN-Gamma; IFN-g; IFNG; IHC; IL-12; IL-13; IL-4 Gene; IL-5 Gene; IL12; IL13; IL4; IL4 gene; IL5; IL5 gene; INFLM; Immune response; Immunohistochemistry; Immunohistochemistry Staining Method; In Vitro; Inflammation; Injury; Interferon Gamma; Interferon Type II; Interferon gamma (human lymphocyte protein moiety reduced); Interferon, Immune; Interferon-gamma; Interleukin 5 (Colony-Stimulating Factor, Eosinophil) Gene; Interleukin 5 Precursor Gene; Interleukin-12; Interleukin-13; Interleukin-4 Gene; Interleukin-4 Precursor Gene; Interleukin-5 Gene; Ito Cell; Ligation; Light; Liver; Liver Fibrosis; Liver diseases; Lymphocyte; Lymphocytic; Magnetism; Maintenance; Maintenances; Mammals, Mice; Mammals, Rodents; Measures; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Methane, tetrachloro-; Mice; Microfluorometry, Flow; Modeling; Mouse Strains; Murine; Mus; NKSF; Natural Killer Cell Stimulatory Factor; Obstruction; Oral; PCR; Pathology; Pattern; Phenotype; Photoradiation; Play; Polymerase Chain Reaction; Population; Preparation; Process; Production; RT-PCR; RTPCR; Regulation; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Rodentia; Rodentias; Role; SCID; SCID Mice; Serum; Severe Combined Immunodeficient Mice; Source; Staging; Stimulus; Subcellular Process; T-Cell Activation; T-Cell Replacing Factor Gene; T-Cells; T-Lymphocyte; TRF Gene; Testing; Tetrachloromethane; Thymus-Dependent Lymphocytes; Time; Tissues; Viral Diseases; Virus Diseases; alcohol induced hepatic injury; alcohol induced liver disorder; alcohol induced liver injury; alcohol-induced hepatic dysfunction; alcohol-induced liver disease; alcohol-induced liver dysfunction; alcohol-mediated liver dysfunction; alcohol-mediated liver injury; alpha-GalCer; alpha-galactosylceramide; balance; balance function; base; bile duct; bile ductule; body system, hepatic; cell sorting; cytokine; ethanol induced hepatic injury; ethanol induced liver disorder; ethanol induced liver injury; ethanol-induced hepatic dysfunction; ethanol-induced liver disease; ethanol-induced liver dysfunction; ethanol-mediated liver dysfunction; ethanol-mediated liver injury; fibrogenesis; flow cytophotometry; hepatic fibrosis; hepatopathy; host response; immunoresponse; inhibitor; inhibitor/antagonist; insight; intrahepatic; killer T cell; lFN-Gamma; liver disorder; liver fibrogenesis; lymph cell; magnetic; mouse model; novel; organ system, hepatic; reconstitute; reconstitution; resistant; response; reverse transcriptase PCR; severe combined immune deficiency; social role; stellate cell; thymus derived lymphocyte; viral infection; virus infection
Project start date: 2009-07-20
Project end date: 2010-06-30
Budget start date: 20-JUL-2009
Budget end date: 30-JUN-2010
PFA/PA: PA-06-001
3K01AA016563-03S1 (2009): $27000
Grants awarded to Ian N Hines
INVOLVEMENT OF NATURAL KILLER T CELLS IN HEPATIC FIBROGENESIS
Ian N Hines
East Carolina University, Office Of Sponsored Programs, Greenville, Nc 27858
Grant 7K01AA016563-04 from National Institute On Alcohol Abuse And Alcoholism
Abstract: Hepatic fibrosis is a consequence of a number of chronic liver pathologies including alcoholic liver disease, viral infection, and biliary outflow obstruction. Growing experimental evidence implicates the class of cytokines produced, specifically the T helper (Th) cytokine phenotype expressed, as a key regulator of the fibrotic response. Indeed, while Th1 type cytokines such as interferon gamma (IFNg) or interleukin (IL) 12 are associated with hepatitis and acute inflammation, expression of the Th2 type cytokines IL4, IL5, and IL13 drive fibrosis development. Moreover, it appears that during chronic liver diseases a switch occurs in which a balanced or perhaps Th1 -shifted cytokine response is replaced by an unbalanced Th2 predominated cytokine phenotype. Natural killer T (NKT) cells make up approximately 50% of the intrahepatic lymphocytes and are capable of producing both Th1 and Th2 type cytokines making them a potential regulator of the hepatic Th cytokine balance. The impact this cell population has on the hepatic fibrotic response is not known. It is hypothesized that NKT cells limit hepatocellular inflammation and promote hepatic fibrosis through production of key regulatory and pro-fibrotic Th2 type cytokines. Utilizing two well described models of hepatic fibrosis in rodents, carbon tetrachloride administration or bile duct ligation, and two mouse models of NKT cell deficiency, CD1d-/- mice and Ja281-/- mice, the role that NKT cells play in the fibrotic response will be evaluated. In the first aim, the direct consequence of a deficiency in hepatic NKT cells in the development of hepatic fibrogenesis will be determined. Furthermore, using in vitro cell coculture, the role of NKT cells and NKT cell derived factors in the activation primary hepatic stellate cells will also be examined. In the second aim, hepatic NKT cell populations will be exhaustively characterized prior to and during the development of hepatic fibrosis focusing on their Th cytokine phenotype. The third aim will extend these findings and determine the role of NKT cell derived cytokines specifically using a novel adoptive transfer approach where NKT cell deficient mice are reconstituted with certain cytokine deficient NKT cells including IFNg deficient and IL4 deficient cells. The final aim will address the role of IL12, a known inhibitor of NKT cell function and survival, in the development and progression of experimental fibrosis. Together, these studies will provide new insight into the mechanisms of T cell-mediated hepatic fibrosis
Keywords: A Mouse; Acute Hepatitis; Address; Adoptive Transfer; Alcoholic Liver Diseases; Antibodies; B Cell Differentiation Factor I Gene; B-Cell Stimulatory Factor 1 Gene; BSF-1 Gene; BSF1 Gene; Bile Ducts; Bile duct structure; Biliary; Blood Serum; Body Tissues; Carbon Tetrachloride; Cell Count; Cell Fraction; Cell Function; Cell Isolation; Cell Number; Cell Process; Cell Segregation; Cell Separation; Cell Separation Technology; Cell physiology; Cell-Extracellular Matrix; Cell/Tissue, Immunohistochemistry; Cells; Cellular Function; Cellular Physiology; Cellular Process; Chronic; Closure by Ligation; Co-culture; Cocultivation; Coculture; Coculture Techniques; Collagen; Contractile Proteins; Cytofluorometry, Flow; Data; Detection; Development; ECM; EDF Gene; Edodekin Alfa; Eosinophil Differentiation Factor Gene; Equilibrium; Extracellular Matrix; Fibrosis; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Gamma interferon; Hepatic; Hepatic Disorder; Hepatic Fibrogenesis; Hepatic Stellate Cell; IFN-Gamma; IFN-g; IFNG; IHC; IL-12; IL-13; IL-4 Gene; IL-5 Gene; IL12; IL13; IL4; IL4 gene; IL5; IL5 gene; INFLM; Immune response; Immunohistochemistry; Immunohistochemistry Staining Method; In Vitro; Inflammation; Injury; Interferon Gamma; Interferon Type II; Interferon gamma (human lymphocyte protein moiety reduced); Interferon, Immune; Interferon-gamma; Interleukin 5 (Colony-Stimulating Factor, Eosinophil) Gene; Interleukin 5 Precursor Gene; Interleukin-12; Interleukin-13; Interleukin-4 Gene; Interleukin-4 Precursor Gene; Interleukin-5 Gene; Ito Cell; Ligation; Light; Liver; Liver Fibrosis; Liver diseases; Lymphocyte; Lymphocytic; Magnetism; Maintenance; Maintenances; Mammals, Mice; Mammals, Rodents; Measures; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Methane, tetrachloro-; Mice; Microfluorometry, Flow; Modeling; Mouse Strains; Murine; Mus; NKSF; Natural Killer Cell Stimulatory Factor; Obstruction; Oral; PCR; Pathology; Pattern; Phenotype; Photoradiation; Play; Polymerase Chain Reaction; Population; Preparation; Process; Production; RT-PCR; RTPCR; Regulation; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Rodentia; Rodentias; Role; SCID; SCID Mice; Serum; Severe Combined Immunodeficient Mice; Source; Staging; Stimulus; Subcellular Process; T cell response; T-Cell Activation; T-Cell Replacing Factor Gene; T-Cells; T-Lymphocyte; TRF Gene; Testing; Tetrachloromethane; Thymus-Dependent Lymphocytes; Time; Tissues; Viral Diseases; Virus Diseases; alcohol induced hepatic injury; alcohol induced liver disorder; alcohol induced liver injury; alcohol-induced hepatic dysfunction; alcohol-induced liver disease; alcohol-induced liver dysfunction; alcohol-mediated liver dysfunction; alcohol-mediated liver injury; alpha-GalCer; alpha-galactosylceramide; balance; balance function; base; bile duct; bile ductule; body system, hepatic; cell sorting; cytokine; ethanol induced hepatic injury; ethanol induced liver disorder; ethanol induced liver injury; ethanol-induced hepatic dysfunction; ethanol-induced liver disease; ethanol-induced liver dysfunction; ethanol-mediated liver dysfunction; ethanol-mediated liver injury; fibrogenesis; flow cytophotometry; hepatic fibrosis; hepatopathy; host response; immunoresponse; inhibitor; inhibitor/antagonist; insight; intrahepatic; killer T cell; lFN-Gamma; liver disorder; liver fibrogenesis; lymph cell; magnetic; mouse model; novel; organ system, hepatic; reconstitute; reconstitution; resistant; response; reverse transcriptase PCR; severe combined immune deficiency; social role; stellate cell; thymus derived lymphocyte; viral infection; virus infection
Project start date: 2007-08-01
Project end date: 2012-06-30
Budget start date: 6-SEP-2010
Budget end date: 30-JUN-2011
PFA/PA: PA-06-001
7K01AA016563-04 (2010): $120609
5K01AA016563-03 (2009): $118445
5K01AA016563-02 (2008): $115786
1K01AA016563-01 (2007): $113207
Liver Stem Cell Response To Inflammation And Alcohol
Ian N Hines
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5F32AA015005-02 from National Institute On Alcohol Abuse And Alcoholism IRG: ZAA1
Abstract: Alcohol-induced liver injury represents a significant etiology of end-stage liver disease characterized inflammation, fat accumulation, and fibrosis. Because traditional treatment options such as organ transplantation have not been very successful, non-traditional therapies including the use of hepatic progenitor cells (HPC) offer an intriguing alternative. HPC proliferation and differentiation seems to be induced by two critical factors, hepatic insufficiency and a decreased ability of mature hepatocytes to replicate. Here, using two models of HSC proliferation, one initiated by inflammation due to Concanavalin A (ConA) and a second by fibrosis due to carbon tetrachloride (CCI4), each followed by partial hepatectomy, we will first determine whether the type of injury affects the differentiation lineage of HPCs. Next, since alcoholic liver disease (ALD) is complicated by fatty liver, we will determine the effect that fat, due to its reduction in the ability of mature hepatocytes to proliferate in response to injury, has on HPC proliferation in these models. It is hypothesized that fat accumulation will promote endogenous HPC proliferation by inhibition of normal hepatocyte function and proliferation within the inflamed or fibrotic and regenerating liver.
Keywords: alcoholic fatty liver, alcoholic hepatitis, alcoholic liver cirrhosis, ethanol, inflammation, liver cell, pathology, stem cell, toxicology, alcoholic beverage consumption, carbon tetrachloride, cell proliferation, concanavalin A, disease /disorder model, fat, fibrosis, flow cytometry, hepatectomy, immunocytochemistry, immunomagnetic separation, laboratory mouse, postdoctoral investigator
Project start date: 2004-04-01
Project end date: 2006-03-31
5F32AA015005-02 (2005): $48296
1F32AA015005-01 (2004): $42976