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Increased expression of Toll-like receptor 3 in intrahepatic biliary epithelial cells at sites of ductular reaction in diseased livers

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Abstract

Background Toll-like receptors (TLRs) may play active roles in both innate and adaptive immune responses in human intrahepatic biliary epithelial cells (HIBECs). The role of TLR3 expressed by HIBECs, however, remains unclear. Methods We determined the in vivo expression of TLRs in biopsy specimens derived from diseased livers immunohistochemically using a panel of monoclonal antibodies against human TLRs. We then examined the response of cultured HIBECs to a TLR3 ligand, polyinosinic–polycytidylic acid (polyI:C). Using siRNAs specific for Toll-IL-1R homology domain-containing adaptor molecule 1 (TICAM-1) and mitochondrial antiviral signaling protein (MAVS), we studied signaling pathways inducing IFN-β expression. Results The expression of TLR3 was markedly increased in biliary epithelial cells at sites of ductular reaction in diseased livers, including primary biliary cirrhosis (PBC), autoimmune hepatitis (AIH), and chronic viral hepatitis (CH) as compared to nondiseased livers. Although cultured HIBECs constitutively expressed TLR3 at both the protein and mRNA levels in vitro, the addition of polyI:C to culture media induced only minimal increases in IFN-β mRNA. In contrast, transfection of HIBECs with polyI:C induced a marked increase in mRNAs encoding a variety of chemokines/cytokines, including IFN-β, IL-6, and TNF-α. The induction of IFN-β mRNA was efficiently inhibited by an siRNA against MAVS but not against TICAM-1, indicating that the main signaling pathway for IFN-β induction following polyI:C transfection is via retinoic acid-inducible gene I (RIG-I)/melanoma differentiation-associated gene 5 (MDA5) in HIBECs. Conclusions TLR3 expression by biliary epithelial cells increased at sites of ductular reaction in diseased livers; further study will be necessary to characterize it’s in vivo physiological role.

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Abbreviations

BEC:

Biliary epithelial cell

CK:

Cytokeratin

dsRNA:

Double stranded RNA

ER:

Endoplasmic reticulum

ELISA:

Enzyme-linked immunosorbent assay

GAPDH:

Glyceraldehydes-3-phosphate dehydrogenase

HIBEC:

Human intrahepatic biliary epithelial cell

HRP:

Horseradish peroxidase

IFN:

Interferon

IL:

Interleukin

IRF:

Interferon regulatory factor

MAVS:

Mitochondrial anti-viral signaling protein

MDA5:

Melanoma differentiation associated gene-5

MyD88:

Myeloid differentiation factor 88

PBC:

Primary biliary cirrhosis

PBMC:

Peripheral blood mononuclear cells

PolyI:C:

Polyinosinic–polycytidylic acid

PRR:

Pattern-recognition receptor

RIG-I:

Retinoic acid-inducible gene I

RT-PCR:

Reverse transcription-polymerase chain reaction

siRNA:

Small interfering RNA

TLR:

Toll-like receptor

TNF:

Tumor necrosis factor

TICAM-1:

Toll-IL-1R homology domain containing adaptor molecule 1

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Acknowledgments

This study was supported by Grants-in-Aid for Scientific Research from the Ministry of Health, Labour and Welfare of Japan and Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Clinical Research Center, National Hospital Organization (NHO) Nagasaki Medical Center, Kubara 2-1001-1, Omura, Nagasaki, 856-8562, Japan

    Minoru Nakamura, Atsumasa Komori, Terufumi Yokoyama, Yoshihiro Aiba, Aiko Araki, Yasushi Takii, Masahiro Ito, Mutsumi Matsuyama, Makiko Koyabu, Kiyoshi Migita, Ken Taniguchi, Hikaru Fujioka, Hiroshi Yatsuhashi & Hiromi Ishibashi

  2. Department of Hepatology, Nagasaki University Graduate School of Biomedical Sciences, Kubara 2-1001-1, Omura, Nagasaki, 856-8562, Japan

    Minoru Nakamura

  3. Department of Microbiology and Immunology, Hokkaido University Graduate School of Medicine, Sapporo, Japan

    Kenji Funami, Misako Matsumoto & Tsukasa Seya

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  1. Minoru Nakamura
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Correspondence to Minoru Nakamura.

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Nakamura, M., Funami, K., Komori, A. et al. Increased expression of Toll-like receptor 3 in intrahepatic biliary epithelial cells at sites of ductular reaction in diseased livers. Hepatol Int 2, 222–230 (2008). https://doi.org/10.1007/s12072-008-9055-4

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  • DOI: https://doi.org/10.1007/s12072-008-9055-4

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