Vascular cell responsiveness to toll-like receptor ligands in carotid atheroma

C. Erridge, A. Burdess, A.J. Jackson, C. Murray, M. Riggio, D. Lappin, S. Milligan, C.M. Spickett, D.J. Webb

Research output: Contribution to journalArticle

Abstract

Background Atherosclerosis is potentiated by stimulation of Toll-like receptors (TLRs), which serve to detect pathogen associated molecular patterns (PAMPs). However little is known of which PAMPs may be present in atheroma, or capable of stimulating inflammatory signalling in vascular cells. Materials and Methods DNA extracted from human carotid atheroma samples was amplified and sequenced using broad-range 16S gene specific primers to establish historical exposure to bacterial PAMPs. Responsiveness of primary human arterial and venous endothelial and smooth muscle cells to PAMPs specific for each of the TLRs was assessed by measurement of interleukin-8 secretion and E-selectin expression. Results Extracts of atheromatous tissue stimulated little or no signalling in TLR-transfected HEK-293 cells. However, sequencing of bacterial DNA amplified from carotid atheroma revealed the presence of DNA from 17 different bacterial genera, suggesting historical exposure to bacterial lipopeptide, lipopolysaccharide and flagellin. All cells examined were responsive to the ligands of TLR3 and TLR4, poly inosine:cytosine and lipopolysaccharide. Arterial cells were responsive to a wider range of PAMPs than venous cells, being additionally responsive to bacterial flagellin and unmethylated cytosine-phosphate-guanosine DNA motifs, the ligands of TLR5 and TLR9, respectively. Cells were generally unresponsive towards the ligands of human TLR7 and TLR8, loxoribine and single stranded RNA. Only coronary artery endothelial cells expressed TLR2 mRNA and responded to the TLR2 ligand Pam3CSK4. Conclusions Vascular cells are responsive to a relatively diverse range of TLR ligands and may be exposed, at least transiently, to ligands of TLR2, TLR4, TLR5 and TLR9 during the development of carotid atheroma.
LanguageEnglish
Pages713-720
Number of pages8
JournalEuropean journal of clinical investigation
Volume38
Issue number10
Early online date20 Sep 2008
DOIs
Publication statusPublished - Oct 2008

Fingerprint

Toll-Like Receptors
Atherosclerotic Plaques
Blood Vessels
Ligands
Flagellin
Cytosine
Lipopolysaccharides
DNA
Lipopeptides
Bacterial DNA
Inosine
Nucleotide Motifs
E-Selectin
Tissue Extracts
Guanine Nucleotides
HEK293 Cells
Endothelial cells
Interleukin-8
Smooth Muscle Myocytes
Muscle

Keywords

  • 16S gene
  • atherosclerosis
  • bacteria
  • toll-like receptor
  • general medicine
  • pharmacology therapeutics
  • pharmacy and materia medica

Cite this

Erridge, C., Burdess, A., Jackson, A. J., Murray, C., Riggio, M., Lappin, D., ... Webb, D. J. (2008). Vascular cell responsiveness to toll-like receptor ligands in carotid atheroma. European journal of clinical investigation, 38(10), 713-720. https://doi.org/10.1111/j.1365-2362.2008.02010.x
Erridge, C. ; Burdess, A. ; Jackson, A.J. ; Murray, C. ; Riggio, M. ; Lappin, D. ; Milligan, S. ; Spickett, C.M. ; Webb, D.J. / Vascular cell responsiveness to toll-like receptor ligands in carotid atheroma. In: European journal of clinical investigation. 2008 ; Vol. 38, No. 10. pp. 713-720.
@article{ebd1d1dc471e491a97017d66e4b95938,
title = "Vascular cell responsiveness to toll-like receptor ligands in carotid atheroma",
abstract = "Background Atherosclerosis is potentiated by stimulation of Toll-like receptors (TLRs), which serve to detect pathogen associated molecular patterns (PAMPs). However little is known of which PAMPs may be present in atheroma, or capable of stimulating inflammatory signalling in vascular cells. Materials and Methods DNA extracted from human carotid atheroma samples was amplified and sequenced using broad-range 16S gene specific primers to establish historical exposure to bacterial PAMPs. Responsiveness of primary human arterial and venous endothelial and smooth muscle cells to PAMPs specific for each of the TLRs was assessed by measurement of interleukin-8 secretion and E-selectin expression. Results Extracts of atheromatous tissue stimulated little or no signalling in TLR-transfected HEK-293 cells. However, sequencing of bacterial DNA amplified from carotid atheroma revealed the presence of DNA from 17 different bacterial genera, suggesting historical exposure to bacterial lipopeptide, lipopolysaccharide and flagellin. All cells examined were responsive to the ligands of TLR3 and TLR4, poly inosine:cytosine and lipopolysaccharide. Arterial cells were responsive to a wider range of PAMPs than venous cells, being additionally responsive to bacterial flagellin and unmethylated cytosine-phosphate-guanosine DNA motifs, the ligands of TLR5 and TLR9, respectively. Cells were generally unresponsive towards the ligands of human TLR7 and TLR8, loxoribine and single stranded RNA. Only coronary artery endothelial cells expressed TLR2 mRNA and responded to the TLR2 ligand Pam3CSK4. Conclusions Vascular cells are responsive to a relatively diverse range of TLR ligands and may be exposed, at least transiently, to ligands of TLR2, TLR4, TLR5 and TLR9 during the development of carotid atheroma.",
keywords = "16S gene, atherosclerosis, bacteria, toll-like receptor, general medicine, pharmacology therapeutics, pharmacy and materia medica",
author = "C. Erridge and A. Burdess and A.J. Jackson and C. Murray and M. Riggio and D. Lappin and S. Milligan and C.M. Spickett and D.J. Webb",
year = "2008",
month = "10",
doi = "10.1111/j.1365-2362.2008.02010.x",
language = "English",
volume = "38",
pages = "713--720",
journal = "European journal of clinical investigation",
issn = "0014-2972",
publisher = "Wiley-Blackwell",
number = "10",

}

Erridge, C, Burdess, A, Jackson, AJ, Murray, C, Riggio, M, Lappin, D, Milligan, S, Spickett, CM & Webb, DJ 2008, 'Vascular cell responsiveness to toll-like receptor ligands in carotid atheroma' European journal of clinical investigation, vol. 38, no. 10, pp. 713-720. https://doi.org/10.1111/j.1365-2362.2008.02010.x

Vascular cell responsiveness to toll-like receptor ligands in carotid atheroma. / Erridge, C.; Burdess, A.; Jackson, A.J.; Murray, C.; Riggio, M.; Lappin, D.; Milligan, S.; Spickett, C.M.; Webb, D.J.

In: European journal of clinical investigation, Vol. 38, No. 10, 10.2008, p. 713-720.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Vascular cell responsiveness to toll-like receptor ligands in carotid atheroma

AU - Erridge, C.

AU - Burdess, A.

AU - Jackson, A.J.

AU - Murray, C.

AU - Riggio, M.

AU - Lappin, D.

AU - Milligan, S.

AU - Spickett, C.M.

AU - Webb, D.J.

PY - 2008/10

Y1 - 2008/10

N2 - Background Atherosclerosis is potentiated by stimulation of Toll-like receptors (TLRs), which serve to detect pathogen associated molecular patterns (PAMPs). However little is known of which PAMPs may be present in atheroma, or capable of stimulating inflammatory signalling in vascular cells. Materials and Methods DNA extracted from human carotid atheroma samples was amplified and sequenced using broad-range 16S gene specific primers to establish historical exposure to bacterial PAMPs. Responsiveness of primary human arterial and venous endothelial and smooth muscle cells to PAMPs specific for each of the TLRs was assessed by measurement of interleukin-8 secretion and E-selectin expression. Results Extracts of atheromatous tissue stimulated little or no signalling in TLR-transfected HEK-293 cells. However, sequencing of bacterial DNA amplified from carotid atheroma revealed the presence of DNA from 17 different bacterial genera, suggesting historical exposure to bacterial lipopeptide, lipopolysaccharide and flagellin. All cells examined were responsive to the ligands of TLR3 and TLR4, poly inosine:cytosine and lipopolysaccharide. Arterial cells were responsive to a wider range of PAMPs than venous cells, being additionally responsive to bacterial flagellin and unmethylated cytosine-phosphate-guanosine DNA motifs, the ligands of TLR5 and TLR9, respectively. Cells were generally unresponsive towards the ligands of human TLR7 and TLR8, loxoribine and single stranded RNA. Only coronary artery endothelial cells expressed TLR2 mRNA and responded to the TLR2 ligand Pam3CSK4. Conclusions Vascular cells are responsive to a relatively diverse range of TLR ligands and may be exposed, at least transiently, to ligands of TLR2, TLR4, TLR5 and TLR9 during the development of carotid atheroma.

AB - Background Atherosclerosis is potentiated by stimulation of Toll-like receptors (TLRs), which serve to detect pathogen associated molecular patterns (PAMPs). However little is known of which PAMPs may be present in atheroma, or capable of stimulating inflammatory signalling in vascular cells. Materials and Methods DNA extracted from human carotid atheroma samples was amplified and sequenced using broad-range 16S gene specific primers to establish historical exposure to bacterial PAMPs. Responsiveness of primary human arterial and venous endothelial and smooth muscle cells to PAMPs specific for each of the TLRs was assessed by measurement of interleukin-8 secretion and E-selectin expression. Results Extracts of atheromatous tissue stimulated little or no signalling in TLR-transfected HEK-293 cells. However, sequencing of bacterial DNA amplified from carotid atheroma revealed the presence of DNA from 17 different bacterial genera, suggesting historical exposure to bacterial lipopeptide, lipopolysaccharide and flagellin. All cells examined were responsive to the ligands of TLR3 and TLR4, poly inosine:cytosine and lipopolysaccharide. Arterial cells were responsive to a wider range of PAMPs than venous cells, being additionally responsive to bacterial flagellin and unmethylated cytosine-phosphate-guanosine DNA motifs, the ligands of TLR5 and TLR9, respectively. Cells were generally unresponsive towards the ligands of human TLR7 and TLR8, loxoribine and single stranded RNA. Only coronary artery endothelial cells expressed TLR2 mRNA and responded to the TLR2 ligand Pam3CSK4. Conclusions Vascular cells are responsive to a relatively diverse range of TLR ligands and may be exposed, at least transiently, to ligands of TLR2, TLR4, TLR5 and TLR9 during the development of carotid atheroma.

KW - 16S gene

KW - atherosclerosis

KW - bacteria

KW - toll-like receptor

KW - general medicine

KW - pharmacology therapeutics

KW - pharmacy and materia medica

UR - http://www.scopus.com/inward/record.url?scp=52149089441&partnerID=8YFLogxK

UR - http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2362.2008.02010.x/abstract

U2 - 10.1111/j.1365-2362.2008.02010.x

DO - 10.1111/j.1365-2362.2008.02010.x

M3 - Article

VL - 38

SP - 713

EP - 720

JO - European journal of clinical investigation

T2 - European journal of clinical investigation

JF - European journal of clinical investigation

SN - 0014-2972

IS - 10

ER -