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From the Laboratories of Molecular
Immunoregulation*
and Experimental
Immunology,
Division of Basic Sciences,
Frederick Cancer Research and Development Center, Frederick, the
Intramural Research Support Program,§
Pathology
Histotechnology Laboratory, SAIC Frederick, Frederick, and the Cell
Biology Section,¶
National Institute of Dental
Research,||
Bethesda, Maryland; and the
Department of Medicine,
St. George Clinical
School, Kogarah, New South Wales, Australia
The contribution of chemokines toward angiogenesis is currently a
focus of intensive investigation. Certain members of the CXC chemokine
family can induce bovine capillary endothelial cell migration in
vitro and corneal angiogenesis in vivo,
and apparently act via binding to their receptors CXCR1 and CXCR2. We
used an RNAse protection assay that permitted the simultaneous
detection of mRNA for various CXC chemokine receptors in resting human
umbilical vein endothelial cells (HUVECs) and detected low levels of
only CXCR4 mRNA. Stimulation of HUVECs with vascular endothelial growth
factor (VEGF) or basic fibroblast growth factor (bFGF) up-regulated
levels of only CXCR4 mRNA. CXCR4 specifically binds the chemokine
stromal-derived factor-1
(SDF-1). Competitive binding studies
using 125I-labeled SDF-1 with Scatchard analysis
indicated that VEGF or bFGF induced an average number of approximately
16,600 CXCR4 molecules per endothelial cell, with a
Kd = 1.23 x 10-9 mol/L.
These receptors were functional as HUVECs and human aorta
endothelial cells (HAECs) migrated toward SDF-1. Although
SDF-1-induced chemotaxis was inhibited by the addition of a
neutralizing monoclonal CXCR4 antibody, endothelial chemotaxis
toward VEGF was not altered; therefore, the angiogenic effect
of VEGF is independent of SDF-1. Furthermore, subcutaneous
SDF-1 injections into mice induced formation of local small blood
vessels that was accompanied by leukocytic infiltrates. To test whether
these effects were dependent on circulating leukocytes, we
successfully obtained SDF-1-induced neovascularization from cross
sections of leukocyte-free rat aorta. Taken together, our data
indicate that SDF-1 acts as a potent chemoattractant for endothelial
cells of different origins bearing CXCR4 and is a participant in
angiogenesis that is regulated at the receptor level by VEGF and
bFGF.
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