Von Willebrand Factor Targets IL-8 to Weibel-Palade Bodies in an Endothelial Cell Line

Exp Cell Res. 2003 May 15;286(1):67-74. doi: 10.1016/s0014-4827(03)00094-6.

Abstract

Vascular endothelial cells are able to store the chemotactic cytokine interleukin-8 (IL-8) in specialized storage vesicles, Weibel-Palade bodies, together with von Willebrand factor (VWF) and P-selectin. We investigated whether VWF plays a role in the sorting of IL-8 into these organelles. We examined the effect of VWF expression on IL-8 targeting in an endothelial cell line (EC-RF24). This cell line has retained the typical phenotypic characteristics of primary endothelial cells but has lost the capacity to produce VWF in appreciable amounts. EC-RF24 cells were retrovirally transduced with a vector encoding a VWF-green fluorescent protein chimera (VWF-GFP). This approach enables direct visualization of the cellular distribution and secretory behavior of the VWF-GFP hybrid. Expression of VWF-GFP resulted in the generation of Weibel-Palade body-like organelles as shown by the colocalization of VWF-GFP and P-selectin. VWF-GFP expressing EC-RF24 cells also showed significant colocalization of VWF-GFP with IL-8 in these storage vesicles. Live cell imaging revealed that the number of VWF-GFP-containing granules decreased upon cell stimulation. These observations indicate that VWF plays an active role in sequestering IL-8 into Weibel-Palade bodies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / metabolism
  • Cathepsin D / metabolism
  • Cell Line
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism*
  • Green Fluorescent Proteins
  • Humans
  • Indicators and Reagents / metabolism
  • Interleukin-1 / metabolism
  • Interleukin-8 / metabolism*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Fluorescence
  • P-Selectin / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Weibel-Palade Bodies / metabolism*
  • von Willebrand Factor / genetics
  • von Willebrand Factor / metabolism*

Substances

  • Antibodies, Monoclonal
  • Indicators and Reagents
  • Interleukin-1
  • Interleukin-8
  • Luminescent Proteins
  • P-Selectin
  • Recombinant Fusion Proteins
  • von Willebrand Factor
  • Green Fluorescent Proteins
  • Cathepsin D