title: Immaturity of microvessels in haemorrhagic plaques is associated with proteolytic degradation of angiogenic factors.
abstract: OBJECTIVE
  We investigated the causes of microvessel immaturity and destabilization in human atherosclerotic lesions.
  
  
  RESULTS
  Human atherosclerotic carotid plaques (n = 24) were classified as non-haemorrhagic (NH) or haemorrhagic (Hem), according to their macroscopic aspect and haemoglobin content. Plaque microvessel density and maturity were quantified by immunohistochemistry. Expression of angiogenic factors was studied by immunohistochemistry, in situ hybridization, and ELISA. Plaque-conditioned media were tested for plasmin and elastase activities and for their ability to degrade angiogenic factors and to induce smooth muscle cell migration. Microvessel density and leucocyte infiltration were increased in Hem compared with NH plaques. Plaque vasculature appeared vulnerable as indicated by the absence of alpha-actin-positive mural cells in most plaque vessels. Despite increased numbers of angiogenic factor-expressing microvessels and leucocytes in Hem plaques, lower levels of vascular endothelial growth factor, placental growth factor, and angiopoietin-1 were found in conditioned media from Hem plaques. However, NH and Hem plaques released similar levels of the vascular destabilizing factor, angiopoietin-2. Addition of recombinant angiogenic factors to plaque extracts showed that all factors but angiopoietin-2 were selectively degraded by plasmin and/or elastase released from Hem plaques. Furthermore, conditioned media from Hem plaques showed a reduced ability to induce smooth muscle cell migration.
  
  
  CONCLUSIONS
  Our results provide evidence that immaturity of plaque vessels is associated with the degradation of angiogenic factors by haemorrhage-conveyed leucocytes and proteases.
authors.affiliation: Inserm and Univ. Paris 7, Cardiovascular Haematology, Bio-Engineering and Remodelling, Bichat-Claude Bernard Hospital, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Le Dall
authors.forename: Benoît
authors.initials: B
authors.lastname: Ho-Tin-Noé
authors.forename: Liliane
authors.initials: L
authors.lastname: Louedec
authors.forename: Olivier
authors.initials: O
authors.lastname: Meilhac
authors.forename: Carmen
authors.initials: C
authors.lastname: Roncal
authors.forename: Peter
authors.initials: P
authors.lastname: Carmeliet
authors.forename: Stéphane
authors.initials: S
authors.lastname: Germain
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Michel
authors.forename: Xavier
authors.initials: X
authors.lastname: Houard
mesh_terms: D000199:Actins / Q000032:analysis; D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D042683:Angiopoietin-1 / Q000378:metabolism; D050197:Atherosclerosis / Q000378:metabolism*; D002465:Cell Movement; D005341:Fibrinolysin / Q000502:physiology; D006470:Hemorrhage / Q000378:metabolism*; D006801:Humans; D019272:Leukocyte Elastase / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism; D055806:Microvessels / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D018919:Neovascularization, Physiologic; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: C467630:ANGPT1 protein, human; D000199:Actins; D043925:Angiogenesis Inducing Agents; D042683:Angiopoietin-1; D008565:Membrane Proteins; C098069:PIGF protein, human; D042461:Vascular Endothelial Growth Factor A; D019272:Leukocyte Elastase; D005341:Fibrinolysin