title: Production of IL-8, VEGF and Elastase by Circulating and Intraplaque Neutrophils in Patients with Carotid Atherosclerosis.
abstract: OBJECTIVE
  Polymorphonuclear neutrophils (PMN) in atherosclerotic plaques have been identified only recently, and their contribution to plaque development is not yet fully understood. In this study, production of elastase, interleukin (IL)-8 and vascular endothelial growth factor (VEGF) by PMN was investigated in subjects with carotid stenosis undergoing carotid endarterectomy (CEA).
  
  
  METHODS
  The study enrolled 50 patients (Pts) and 10 healthy subjects (HS). Circulating PMN (cPMN) isolated from venous blood (in both Pts and HS) and from plaques (pPMN, in Pts) were cultured, alone or with 0.1 μM fMLP. Elastase, IL-8 and VEGF mRNA were analyzed by real-time PCR. In CEA specimens, PMN were localized by immunohistochemistry.
  
  
  RESULTS
  In both Pts cPMN and pPMN, IL-8 mRNA was higher at rest but lower after fMLP (P<0.01 vs HS), and VEGF mRNA was higher both at rest and after fMLP (P<0.01 vs HS), while elastase mRNA was not significantly different. On the contrary, protein production was always higher in cPMN of HS with respect to values measured in cells of Pts. In CEA specimens, CD66b+ cells localized to areas with massive plaque formation close to neovessels. Pts with soft and mix plaques, as defined by computed tomography, did not differ in cPMN or pPMN IL-8, VEGF or elastase mRNA, or in intraplaque CD66b+ cell density. However, Pts with soft plaques had higher white blood cell count due to increased PMN.
  
  
  CONCLUSIONS
  In Pts with carotid plaques, both circulating and intraplaque PMN produce IL-8, VEGF and elastase, which are crucial for plaque development and progression. These findings suggest mechanistic explanations to the reported correlation between PMN count and cardiovascular mortality in carotid ATH.
authors.affiliation: Center of Research in Medical Pharmacology, University of Insubria, Varese, Italy.
authors.forename: Franca
authors.initials: F
authors.lastname: Marino
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Tozzi
authors.affiliation: Center of Research in Medical Pharmacology, University of Insubria, Varese, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Schembri
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Ferraro
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Antonino
authors.initials: A
authors.lastname: Tarallo
authors.affiliation: Center of Research in Medical Pharmacology, University of Insubria, Varese, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Scanzano
authors.affiliation: Center of Research in Medical Pharmacology, University of Insubria, Varese, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Legnaro
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Patrizio
authors.initials: P
authors.lastname: Castelli
authors.affiliation: Center of Research in Medical Pharmacology, University of Insubria, Varese, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Cosentino
mesh_terms: D000368:Aged; D015703:Antigens, CD / Q000378:metabolism; D002340:Carotid Artery Diseases / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002465:Cell Movement; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D009504:Neutrophils / Q000378:metabolism*; D010196:Pancreatic Elastase / Q000096:biosynthesis* / Q000235:genetics; D058226:Plaque, Atherosclerotic / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D015703:Antigens, CD; C069847:CD66 antigens; C508606:CXCL8 protein, human; D015815:Cell Adhesion Molecules; D016209:Interleukin-8; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D010196:Pancreatic Elastase