Expression and regulation of Toll-like receptor 2 by IL-1beta and fibronectin fragments in human articular chondrocytes

S-L Su; C-D Tsai; C-H Lee; D M Salter; H-S Lee

doi:10.1016/j.joca.2005.04.017

Expression and regulation of Toll-like receptor 2 by IL-1beta and fibronectin fragments in human articular chondrocytes

S-L Su, C-D Tsai, C-H Lee, D M Salter, H-S Lee

Research output: Contribution to journal › Article › peer-review

Abstract / Description of output

OBJECTIVE: The objective of this study was to examine expression and regulation of Toll-like receptor 2 (TLR2) in human articular chondrocytes.

METHODS: Human articular chondrocytes were enzymatically isolated from normal and osteoarthritic knee cartilage. Immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to assess the expression of toll-like receptors. Following stimulation of chondrocytes in vitro by IL-1beta and fibronectin proteolytic fragments, the relative levels of mRNA for TLR2 were determined by quantitative real-time PCR. MyD88 activation and nuclear factor-kappaB (NF-kappaB) translocation were evaluated by immunoprecipitation and electrophoretic mobility shift assay, respectively.

RESULTS: Human articular chondrocytes mainly expressed TLR1, 2, 5 by RT-PCR. Protein expression of TLR2 was also identified in adult human articular cartilage. TLR2 was upregulated following IL-1beta and fibronectin proteolytic fragments stimulation in primary cultures of osteoarthritic articular chondrocytes. Fibronectin proteolytic fragments-induced TLR2 upregulation involved an IL-1beta autocrine/paracrine pathway.

CONCLUSIONS: TLR2 is expressed in human articular cartilage and is upregulated by proarthritic agents including IL-1beta and fibronectin fragments. Signaling through TLR is a novel pro-inflammatory mechanism in osteoarthritis and targeting of these signaling pathways may be of value in treatment of degenerative joint disease.

Original language	English
Pages (from-to)	879-86
Number of pages	8
Journal	Osteoarthritis and Cartilage
Volume	13
Issue number	10
DOIs	https://doi.org/10.1016/j.joca.2005.04.017
Publication status	Published - Oct 2005

Keywords / Materials (for Non-textual outputs)

Aged
Blotting, Western
Cartilage, Articular
Cells, Cultured
Chondrocytes
Fibronectins
Gene Expression Regulation
Humans
Interleukin-1
Middle Aged
Osteoarthritis, Knee
Peptide Fragments
RNA, Messenger
Reverse Transcriptase Polymerase Chain Reaction
Signal Transduction
Toll-Like Receptor 2

Access to Document

10.1016/j.joca.2005.04.017

Cite this

@article{fb4f5d6b198b4eb8804ac2812a7755f0,

title = "Expression and regulation of Toll-like receptor 2 by IL-1beta and fibronectin fragments in human articular chondrocytes",

abstract = "OBJECTIVE: The objective of this study was to examine expression and regulation of Toll-like receptor 2 (TLR2) in human articular chondrocytes.METHODS: Human articular chondrocytes were enzymatically isolated from normal and osteoarthritic knee cartilage. Immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to assess the expression of toll-like receptors. Following stimulation of chondrocytes in vitro by IL-1beta and fibronectin proteolytic fragments, the relative levels of mRNA for TLR2 were determined by quantitative real-time PCR. MyD88 activation and nuclear factor-kappaB (NF-kappaB) translocation were evaluated by immunoprecipitation and electrophoretic mobility shift assay, respectively.RESULTS: Human articular chondrocytes mainly expressed TLR1, 2, 5 by RT-PCR. Protein expression of TLR2 was also identified in adult human articular cartilage. TLR2 was upregulated following IL-1beta and fibronectin proteolytic fragments stimulation in primary cultures of osteoarthritic articular chondrocytes. Fibronectin proteolytic fragments-induced TLR2 upregulation involved an IL-1beta autocrine/paracrine pathway.CONCLUSIONS: TLR2 is expressed in human articular cartilage and is upregulated by proarthritic agents including IL-1beta and fibronectin fragments. Signaling through TLR is a novel pro-inflammatory mechanism in osteoarthritis and targeting of these signaling pathways may be of value in treatment of degenerative joint disease.",

keywords = "Aged, Blotting, Western, Cartilage, Articular, Cells, Cultured, Chondrocytes, Fibronectins, Gene Expression Regulation, Humans, Interleukin-1, Middle Aged, Osteoarthritis, Knee, Peptide Fragments, RNA, Messenger, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction, Toll-Like Receptor 2",

author = "S-L Su and C-D Tsai and C-H Lee and Salter, {D M} and H-S Lee",

year = "2005",

month = oct,

doi = "10.1016/j.joca.2005.04.017",

language = "English",

volume = "13",

pages = "879--86",

journal = "Osteoarthritis and Cartilage",

issn = "1063-4584",

publisher = "W.B. Saunders Company",

number = "10",

}

TY - JOUR

T1 - Expression and regulation of Toll-like receptor 2 by IL-1beta and fibronectin fragments in human articular chondrocytes

AU - Su, S-L

AU - Tsai, C-D

AU - Lee, C-H

AU - Salter, D M

AU - Lee, H-S

PY - 2005/10

Y1 - 2005/10

N2 - OBJECTIVE: The objective of this study was to examine expression and regulation of Toll-like receptor 2 (TLR2) in human articular chondrocytes.METHODS: Human articular chondrocytes were enzymatically isolated from normal and osteoarthritic knee cartilage. Immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to assess the expression of toll-like receptors. Following stimulation of chondrocytes in vitro by IL-1beta and fibronectin proteolytic fragments, the relative levels of mRNA for TLR2 were determined by quantitative real-time PCR. MyD88 activation and nuclear factor-kappaB (NF-kappaB) translocation were evaluated by immunoprecipitation and electrophoretic mobility shift assay, respectively.RESULTS: Human articular chondrocytes mainly expressed TLR1, 2, 5 by RT-PCR. Protein expression of TLR2 was also identified in adult human articular cartilage. TLR2 was upregulated following IL-1beta and fibronectin proteolytic fragments stimulation in primary cultures of osteoarthritic articular chondrocytes. Fibronectin proteolytic fragments-induced TLR2 upregulation involved an IL-1beta autocrine/paracrine pathway.CONCLUSIONS: TLR2 is expressed in human articular cartilage and is upregulated by proarthritic agents including IL-1beta and fibronectin fragments. Signaling through TLR is a novel pro-inflammatory mechanism in osteoarthritis and targeting of these signaling pathways may be of value in treatment of degenerative joint disease.

AB - OBJECTIVE: The objective of this study was to examine expression and regulation of Toll-like receptor 2 (TLR2) in human articular chondrocytes.METHODS: Human articular chondrocytes were enzymatically isolated from normal and osteoarthritic knee cartilage. Immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to assess the expression of toll-like receptors. Following stimulation of chondrocytes in vitro by IL-1beta and fibronectin proteolytic fragments, the relative levels of mRNA for TLR2 were determined by quantitative real-time PCR. MyD88 activation and nuclear factor-kappaB (NF-kappaB) translocation were evaluated by immunoprecipitation and electrophoretic mobility shift assay, respectively.RESULTS: Human articular chondrocytes mainly expressed TLR1, 2, 5 by RT-PCR. Protein expression of TLR2 was also identified in adult human articular cartilage. TLR2 was upregulated following IL-1beta and fibronectin proteolytic fragments stimulation in primary cultures of osteoarthritic articular chondrocytes. Fibronectin proteolytic fragments-induced TLR2 upregulation involved an IL-1beta autocrine/paracrine pathway.CONCLUSIONS: TLR2 is expressed in human articular cartilage and is upregulated by proarthritic agents including IL-1beta and fibronectin fragments. Signaling through TLR is a novel pro-inflammatory mechanism in osteoarthritis and targeting of these signaling pathways may be of value in treatment of degenerative joint disease.

KW - Aged

KW - Blotting, Western

KW - Cartilage, Articular

KW - Cells, Cultured

KW - Chondrocytes

KW - Fibronectins

KW - Gene Expression Regulation

KW - Humans

KW - Interleukin-1

KW - Middle Aged

KW - Osteoarthritis, Knee

KW - Peptide Fragments

KW - RNA, Messenger

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Signal Transduction

KW - Toll-Like Receptor 2

U2 - 10.1016/j.joca.2005.04.017

DO - 10.1016/j.joca.2005.04.017

M3 - Article

C2 - 15961329

SN - 1063-4584

VL - 13

SP - 879

EP - 886

JO - Osteoarthritis and Cartilage

JF - Osteoarthritis and Cartilage

IS - 10

ER -

Expression and regulation of Toll-like receptor 2 by IL-1beta and fibronectin fragments in human articular chondrocytes

Abstract / Description of output

Keywords / Materials (for Non-textual outputs)

Access to Document

Fingerprint

Cite this