title: Certolizumab pegol attenuates the pro-inflammatory state in endothelial cells in a manner that is atheroprotective.
abstract: OBJECTIVE
  Rheumatoid arthritis (RA) is associated with accelerated atherosclerosis and premature cardiovascular death. Anti-TNF therapy is thought to reduce clinical cardiovascular disease risk and improve vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to explore the effects of certolizumab pegol (CZP) on TNF-activated human aortic endothelial cells (HAoECs).
  
  
  METHODS
  HAoECs were cultured in vitro and exposed to i) TNF alone, ii) TNF plus CZP, or iii) neither agent. Microarray analysis and quantitative polymerase chain reaction were used to analyse gene expression. Activation of NF-κB was investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was performed to detect microparticle release from HAoECs.
  
  
  RESULTS
  TNF alone had strong effects on endothelial gene expression, while TNF and CZP together produced a global gene expression pattern similar to untreated controls. In particular, genes for E-selectin, VCAM-1 and ICAM-1 were significantly up-regulated by TNF treatment. Notably, the TNF/CZP cocktail prevented the up-regulation of these genes. TNF-induced nuclear translocation of NF-κB was abolished by treatment with CZP. In addition the increased production of endothelial microparticles in TNF-activated HAoECs was prevented by treatment with CZP.
  
  
  CONCLUSIONS
  We have found at cellular level, that a clinically available TNF inhibitor, CZP i) reduces adhesion molecule expression; ii) prevents TNF-induced activation of the NF-κB pathway and iii) prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions.
authors.affiliation: University of Manchester, UK. sarahheathfield@hotmail.com
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Heathfield
authors.forename: Benjamin
authors.initials: B
authors.lastname: Parker
authors.forename: Leo A H
authors.initials: LA
authors.lastname: Zeef
authors.forename: Ian N
authors.initials: IN
authors.lastname: Bruce
authors.forename: M Yvonne
authors.initials: MY
authors.lastname: Alexander
mesh_terms: D021581:Active Transport, Cell Nucleus; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology*; D015153:Blotting, Western; D055252:Cell-Derived Microparticles / Q000187:drug effects / Q000276:immunology; D002478:Cells, Cultured; D000068582:Certolizumab Pegol; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000276:immunology; D005434:Flow Cytometry; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology*; D007150:Immunohistochemistry; D007249:Inflammation / Q000235:genetics / Q000276:immunology / Q000517:prevention & control*; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D011092:Polyethylene Glycols / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D061067:Antibodies, Monoclonal, Humanized; D019040:E-Selectin; D020672:I-kappa B Proteins; C573420:ICAM1 protein, human; D007140:Immunoglobulin Fab Fragments; D016328:NF-kappa B; C000606204:NFKBIA protein, human; C546936:SELE protein, human; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D000072000:NF-KappaB Inhibitor alpha; D011092:Polyethylene Glycols; D000068582:Certolizumab Pegol