Hepatic stellate cells primed with cytokines upregulate inflammation in response to peptidoglycan or lipoteichoic acid

Yong Han Paik, Kwan Sik Lee, Hyun Jin Lee, Kyung Min Yang, Se Jun Lee, Dong Ki Lee, Kwang Hyub Han, Chae Yoon Chon, Sang In Lee, Young Myoung Moon, David A. Brenner

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Gram-positive bacterial products such as peptidoglycan (PGN) and lipoteichoic acid (LTA) are potent stimulators of innate inflammatory responses. We previously reported that lipopolysaccharide (LPS), a major biologically active agent of gram-negative bacteria, induces a proinflammatory response via the Toll-like receptor (TLR) 4 in hepatic stellate cells (HSCs). Here we investigated the mechanism of proinflammatory action by PGN and LTA in activated human HSCs. Following treatment with either TNF-α or IL-1β, expression of TLR2 and CD14 was determined by real-time PCR and Western blotting. NF-κB activation was assessed by NF-κB-driven luciferase assay and electrophoretic mobility shift assay. Interleukin-8 (IL-8) from culture supernatant was measured by ELISA. Activated human HSCs express TLR2 and CD14, which are receptors for PGN and LTA signaling. TNF-α and IL-1β significantly upregulated the expression of TLR2 mRNA and protein in HSCs. PGN and LTA induced NF-κB activation and stimulated production of IL-8 in HSCs. Pretreatment with TNF-α or IL-1β augmented NF-κB activation and IL-8 production in response to PGN or LTA. Both PGN- and LTA-induced NF-κB activation and IL-8 secretion were completely inhibited by anti-TLR2 blocking antibody (T2.5). These findings suggest that TNF-α or IL-1β primed HSCs enhance the production of IL-8 in response to PGN and LTA through augmentation of the TLR2 system.

Original languageEnglish
Pages (from-to)676-686
Number of pages11
JournalLaboratory Investigation
Volume86
Issue number7
DOIs
Publication statusPublished - 2006 Jul 1

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Hepatic Stellate Cells
Peptidoglycan
Interleukin-8
Up-Regulation
Cytokines
Inflammation
Interleukin-1
Toll-Like Receptor 4
Blocking Antibodies
Electrophoretic Mobility Shift Assay
Gram-Negative Bacteria
Luciferases
Lipopolysaccharides
lipoteichoic acid
Real-Time Polymerase Chain Reaction
Western Blotting
Enzyme-Linked Immunosorbent Assay
Messenger RNA
Proteins

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

Cite this

Paik, Yong Han ; Lee, Kwan Sik ; Lee, Hyun Jin ; Yang, Kyung Min ; Lee, Se Jun ; Lee, Dong Ki ; Han, Kwang Hyub ; Chon, Chae Yoon ; Lee, Sang In ; Moon, Young Myoung ; Brenner, David A. / Hepatic stellate cells primed with cytokines upregulate inflammation in response to peptidoglycan or lipoteichoic acid. In: Laboratory Investigation. 2006 ; Vol. 86, No. 7. pp. 676-686.
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abstract = "Gram-positive bacterial products such as peptidoglycan (PGN) and lipoteichoic acid (LTA) are potent stimulators of innate inflammatory responses. We previously reported that lipopolysaccharide (LPS), a major biologically active agent of gram-negative bacteria, induces a proinflammatory response via the Toll-like receptor (TLR) 4 in hepatic stellate cells (HSCs). Here we investigated the mechanism of proinflammatory action by PGN and LTA in activated human HSCs. Following treatment with either TNF-α or IL-1β, expression of TLR2 and CD14 was determined by real-time PCR and Western blotting. NF-κB activation was assessed by NF-κB-driven luciferase assay and electrophoretic mobility shift assay. Interleukin-8 (IL-8) from culture supernatant was measured by ELISA. Activated human HSCs express TLR2 and CD14, which are receptors for PGN and LTA signaling. TNF-α and IL-1β significantly upregulated the expression of TLR2 mRNA and protein in HSCs. PGN and LTA induced NF-κB activation and stimulated production of IL-8 in HSCs. Pretreatment with TNF-α or IL-1β augmented NF-κB activation and IL-8 production in response to PGN or LTA. Both PGN- and LTA-induced NF-κB activation and IL-8 secretion were completely inhibited by anti-TLR2 blocking antibody (T2.5). These findings suggest that TNF-α or IL-1β primed HSCs enhance the production of IL-8 in response to PGN and LTA through augmentation of the TLR2 system.",
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Hepatic stellate cells primed with cytokines upregulate inflammation in response to peptidoglycan or lipoteichoic acid. / Paik, Yong Han; Lee, Kwan Sik; Lee, Hyun Jin; Yang, Kyung Min; Lee, Se Jun; Lee, Dong Ki; Han, Kwang Hyub; Chon, Chae Yoon; Lee, Sang In; Moon, Young Myoung; Brenner, David A.

In: Laboratory Investigation, Vol. 86, No. 7, 01.07.2006, p. 676-686.

Research output: Contribution to journalArticle

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AU - Paik, Yong Han

AU - Lee, Kwan Sik

AU - Lee, Hyun Jin

AU - Yang, Kyung Min

AU - Lee, Se Jun

AU - Lee, Dong Ki

AU - Han, Kwang Hyub

AU - Chon, Chae Yoon

AU - Lee, Sang In

AU - Moon, Young Myoung

AU - Brenner, David A.

PY - 2006/7/1

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N2 - Gram-positive bacterial products such as peptidoglycan (PGN) and lipoteichoic acid (LTA) are potent stimulators of innate inflammatory responses. We previously reported that lipopolysaccharide (LPS), a major biologically active agent of gram-negative bacteria, induces a proinflammatory response via the Toll-like receptor (TLR) 4 in hepatic stellate cells (HSCs). Here we investigated the mechanism of proinflammatory action by PGN and LTA in activated human HSCs. Following treatment with either TNF-α or IL-1β, expression of TLR2 and CD14 was determined by real-time PCR and Western blotting. NF-κB activation was assessed by NF-κB-driven luciferase assay and electrophoretic mobility shift assay. Interleukin-8 (IL-8) from culture supernatant was measured by ELISA. Activated human HSCs express TLR2 and CD14, which are receptors for PGN and LTA signaling. TNF-α and IL-1β significantly upregulated the expression of TLR2 mRNA and protein in HSCs. PGN and LTA induced NF-κB activation and stimulated production of IL-8 in HSCs. Pretreatment with TNF-α or IL-1β augmented NF-κB activation and IL-8 production in response to PGN or LTA. Both PGN- and LTA-induced NF-κB activation and IL-8 secretion were completely inhibited by anti-TLR2 blocking antibody (T2.5). These findings suggest that TNF-α or IL-1β primed HSCs enhance the production of IL-8 in response to PGN and LTA through augmentation of the TLR2 system.

AB - Gram-positive bacterial products such as peptidoglycan (PGN) and lipoteichoic acid (LTA) are potent stimulators of innate inflammatory responses. We previously reported that lipopolysaccharide (LPS), a major biologically active agent of gram-negative bacteria, induces a proinflammatory response via the Toll-like receptor (TLR) 4 in hepatic stellate cells (HSCs). Here we investigated the mechanism of proinflammatory action by PGN and LTA in activated human HSCs. Following treatment with either TNF-α or IL-1β, expression of TLR2 and CD14 was determined by real-time PCR and Western blotting. NF-κB activation was assessed by NF-κB-driven luciferase assay and electrophoretic mobility shift assay. Interleukin-8 (IL-8) from culture supernatant was measured by ELISA. Activated human HSCs express TLR2 and CD14, which are receptors for PGN and LTA signaling. TNF-α and IL-1β significantly upregulated the expression of TLR2 mRNA and protein in HSCs. PGN and LTA induced NF-κB activation and stimulated production of IL-8 in HSCs. Pretreatment with TNF-α or IL-1β augmented NF-κB activation and IL-8 production in response to PGN or LTA. Both PGN- and LTA-induced NF-κB activation and IL-8 secretion were completely inhibited by anti-TLR2 blocking antibody (T2.5). These findings suggest that TNF-α or IL-1β primed HSCs enhance the production of IL-8 in response to PGN and LTA through augmentation of the TLR2 system.

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