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Vol. 9, Issue 4, 875-884, April 1998
*Department of Dermatology, Virchow Clinics, Humboldt
Universität zu Berlin, 13353 Berlin, Germany; and
Mast cells have been implicated in various diseases that are
accompanied by neovascularization. The exact mechanisms by which mast
cells might mediate an angiogenic response, however, are unclear and
therefore, we have investigated the possible expression of vascular
endothelial growth factor/vascular permeability factor (VEGF/VPF) in
the human mast cell line HMC-1 and in human skin mast cells. Reverse
transcription-polymerase chain reaction (RT-PCR) analysis revealed that
mast cells constitutively express VEGF121, VEGF165, and VEGF189. After a prolonged
stimulation of cells for 24 h with phorbol 12-myristate 13-acetate
(PMA) and the ionophore A23187, an additional transcript representing
VEGF206 was detectable, as could be verified by sequence
analysis. These results were confirmed at the protein level by Western
blot analysis. When the amounts of VEGF released under unstimulated and
stimulated conditions were compared, a significant increase was
detectable after stimulation of cells. Human microvascular endothelial
cells (HMVEC) responded to the supernatant of unstimulated HMC-1 cells with a dose-dependent mitogenic effect, neutralizable up to 90% in the
presence of a VEGF-specific monoclonal antibody. Flow cytometry and
postembedding immunoelectron microscopy were used to detect VEGF in its
cell-associated form. VEGF was exclusively detectable in the secretory
granules of isolated human skin mast cells. These results show that
both normal and leukemic human mast cells constitutively express
bioactive VEGF. Furthermore, this study contributes to the
understanding of the physiological role of the strongly heparin-binding VEGF isoforms, since these were found for the first time to be expressed in an activation-dependent manner in HMC-1 cells.
German Institute of Nutrition (DIfE),
Potsdam-Rehbrücke, Potsdam, Germany
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