The phenotype of infiltrating macrophages influences arteriosclerotic plaque vulnerability in the carotid artery

J Stroke Cerebrovasc Dis. 2013 Oct;22(7):910-8. doi: 10.1016/j.jstrokecerebrovasdis.2012.11.020. Epub 2012 Dec 27.

Abstract

Background: Proinflammatory (M1) macrophages and anti-inflammatory (M2) macrophages have been identified in atherosclerotic plaques. While these macrophages have been speculated to be related to plaque vulnerability, there are limited studies investigating this relationship. Therefore, we examined the association between macrophage phenotype (M1 versus M2) and plaque vulnerability and clinical events.

Methods: Patients undergoing carotid endarterectomy received an ultrasound of the carotid artery before surgery. Plaques were processed for analysis by immunohistochemistry, Western blotting, and real-time polymerase chain reaction studies. Medical history and clinical data were obtained from medical records.

Results: Patients were divided into 2 groups: those suffering from acute ischemic attack (symptomatic, n = 31) and those that did not present with symptoms (asymptomatic, n = 34). Ultrasound analysis revealed that plaque vulnerability was greater in the symptomatic group (P= .033; Chi-square test). Immunohistochemistry revealed that plaques from the symptomatic group had a greater concentration of M1 macrophages (CD68-, CD11c-positive) while plaques from the asymptomatic group had more M2 macrophages (CD163-positive). This observation was confirmed by Western blotting. Characterization by real-time polymerase chain reaction studies revealed that plaques from the symptomatic group had increased expression of the M1 markers CD68 and CD11c, as well as monocyte chemoattractive protein-1, interleukin-6, and matrix metalloproteinase-9. In addition, more M1 macrophages expressed in unstable plaques were defined by ultrasound analysis, while more M2 macrophages were expressed in stable plaques.

Conclusions: Our data show that M1 macrophage content of atherosclerotic plaques is associated with clinical incidence of ischemic stroke and increased inflammation or fibrinolysis. We also show the benefits of using ultrasound to evaluate vulnerability in the plaques.

Keywords: Atherosclerosis; carotid artery disease; inflammation; macrophage; stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Antigens, CD / metabolism
  • Antigens, Differentiation, Myelomonocytic / metabolism
  • CD11c Antigen / metabolism
  • Carotid Arteries / diagnostic imaging
  • Carotid Arteries / metabolism*
  • Carotid Arteries / surgery
  • Endarterectomy, Carotid
  • Female
  • Humans
  • Interleukin-6 / metabolism
  • Macrophages / metabolism*
  • Male
  • Matrix Metalloproteinase 9 / metabolism
  • Middle Aged
  • Plaque, Atherosclerotic / diagnostic imaging
  • Plaque, Atherosclerotic / metabolism*
  • Plaque, Atherosclerotic / surgery
  • Ultrasonography

Substances

  • Antigens, CD
  • Antigens, Differentiation, Myelomonocytic
  • CD11c Antigen
  • CD68 antigen, human
  • Interleukin-6
  • Matrix Metalloproteinase 9