Immune stimulation of hepatic fibrogenesis by CD8 cells and attenuation by transgenic interleukin-10 from hepatocytes

doi:10.1053/j.gastro.2004.04.062

Gastroenterology

Volume 127, Issue 3, September 2004, Pages 870-882

https://doi.org/10.1053/j.gastro.2004.04.062 Get rights and content

Backgrounds & Aims: Immunomodulatory cytokines, including interleukin-10 (IL-10), may mediate hepatic fibrosis. Methods: We generated transgenic (TG) mice with hepatocyte expression of rat IL-10 (rIL-10) to assess its impact on lymphocyte subsets and activation of hepatic stellate cells following liver injury from carbon tetrachloride (CCl₄) or thioacetamide (TAA). Results: Fibrosis was reduced in the TG animals in both models, which was not explained solely by differences in liver injury. By fluorescence-activated cell sorter (FACS), there were less CD4+ T cells in naive TG mice, and, following fibrosis induction, CD4+ T cells decreased only in wild-type (WT) mice, whereas increases in CD8+ T cells seen in WT animals were significantly attenuated in TG mice. Subtotal irradiation diminished fibrosis equally in both WT and TG groups, suggesting that rIL-10’s antifibrotic effect was lymphocyte mediated. To assess the role of lymphocytes on stellate cell activation, either whole splenic lymphocytes, CD4+, or CD8+ T-cell subsets from WT animals with CCl₄ fibrosis were adoptively transferred to severe combined immunodeficiency (SCID) recipients, which led to stellate cell activation and fibrogenic stimulation as assessed by expression of transforming growth factor (TGF)-β1 and collagen I messenger RNA (mRNA) and by immunoblot of α-smooth muscle actin. Moreover, serum aminotransferase levels and stellate cell activation mRNA were significantly higher among the CD8+ T-cell recipients. Conclusions: Transgenic expression of rIL-10 in liver leads to reduced fibrosis and alterations in liver lymphocyte subsets both in untreated liver and following fibrosis induction. In this model, fibrosis may be a CD8+ T-cell-mediated disease that is attenuated by rIL-10.

Section snippets

Animals

C57BL/6 mice (wild-type and SCID) were purchased from Jackson Laboratories. SCID mice were housed in a barrier facility. Animals received care according to National Institutes of Health guidelines.

Generation and characterization of rat IL-10 TG mice

TG mice expressing rIL-10 primarily in hepatocytes were generated by expressing the full-length rIL-10 cDNA (accession no. L02926, nucleotides 1–682) downstream of the transthyretin (TTR) promoter on a mixed C57BL/6 and C3H background, using an expression construct generously provided by Dr. T.

Transgenic rIL-10 is antifibrotic in 2 models of liver injury

We analyzed fibrosis in WT and TG mice following induction of fibrosis in 2 mechanistically distinct models of injury due to CCl₄ and TAA, respectively. Serum rIL-10 levels were undetectable in all WT animals from both groups and 147 ± 222 pg/mL in the untreated TG animals, whereas concentrations in the TG CCl₄ and TAA groups were 182.6 ± 101.5 and 292.6 ±280 pg/mL, respectively.

In the WT CCl₄ group the fibrotic area based on Bioquant morphometry was 4.3% (±2.5%) vs. TG 1.5% (±0.85%) (P < 0.03)

Discussion

Our study has explored the potential interactions of humoral factors and lymphocyte subsets by first assessing the impact of hepatic transgenic rIL-10 on liver fibrosis and lymphocyte responses in vivo then exploiting this response to dissect the contributions of lymphocyte subsets on fibrogenesis. IL-10 was chosen because of its broad immunomodulatory and antifibrotic properties, providing a useful reagent to perturb the homeostasis of normal and injured liver. IL-10 is a regulatory cytokine

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: Supported by grants from NIDDK (DK37340 and DK56621 to S.L.F. and DK02965-01 to W.Z.M.), Feld Family Trust, Artzt Primary Biliary Cirrhosis Program, and Shirley Fiterman Basic Science Research Award (to W.Z.M.).

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Basic-liver, pancreas, and biliary tractImmune stimulation of hepatic fibrogenesis by CD8 cells and attenuation by transgenic interleukin-10 from hepatocytes

Section snippets

Animals

Generation and characterization of rat IL-10 TG mice

Transgenic rIL-10 is antifibrotic in 2 models of liver injury

Discussion

J Biol Chem

J Biol Chem

Gastroenterology

Hepatology

Gastroenterology

Hepatology

J Hepatol

Hepatology

J Hepatol

Blood

Hepatology

Gastroenterology

Hepatology

Liver fibrosis—from bench to bedside

J Hepatol

Roles of TGF-β in hepatic fibrosis

Front Biosci

Expression of connective tissue growth factor in experimental rat and human liver fibrosis

Hepatology

IL-10 receptor and coreceptor expression in quiescent and activated hepatic stellate cells

Am J Physiol Gastrointest Liver Physiol

Interleukin-10 expression and function in experimental murine liver inflammation and fibrosis

Hepatology

Liver injury and fibrosis in interferon gamma knockout mice

Hepatology

Production and role of interleukin-10 in concanavalin A-induced hepatitis in mice

Hepatology

Interleukin-10 controls neutrophilic infiltration, hepatocyte proliferation, and liver fibrosis induced by carbon tetrachloride in mice

Hepatology

Reciprocal expression of interferon γ or interleukin 4 during the resolution or progression of murine leishmaniasis. Evidence for expansion of distinct helper T-cell subsets

J Exp Med

Strain-specific differences in mouse hepatic wound healing are mediated by divergent T-helper cytokine responses

Proc Natl Acad Sci U S A

Interferon γ-independent effects of interleukin 12 administered during acute or established infection due to Leishmania major

Proc Natl Acad Sci U S A

CD4+ effector cells default to the Th2 pathway in interferon γ-deficient mice infected with Leishmania major

J Exp Med

Influence of human immunodeficiency virus type 1 infection on the natural course of chronic parenterally acquired hepatitis C

Eur J Clin Microbiol Infect Dis

Liver fibrosis progression in human immunodeficiency virus and hepatitis C virus coinfected patients. The Multivirc Group

Hepatology

Fibrosis and disease progression in hepatitis C

Hepatology

Targeted in vivo expression of the cyclin-dependent kinase inhibitor p21 halts hepatocyte cell-cycle progression, postnatal liver development and regeneration

Genes Dev

Basic-liver, pancreas, and biliary tract
Immune stimulation of hepatic fibrogenesis by CD8 cells and attenuation by transgenic interleukin-10 from hepatocytes