Induction of apoptosis in human dermal microvascular endothelial cells and infantile hemangiomas by interferon-alpha.
BACKGROUND
Hemangioma of infancy is an angiomatous disorder characterized by the proliferation of capillary endothelium. It has been shown that interferon-alpha (IFN-alpha) may induce involution of proliferating, life-threatening hemangiomas in children. This IFN-alpha-induced regression of hemangiomas is not accompanied by any T cell response nor by the occurrence of necrosis.


METHODS
To determine whether IFN-alpha may induce apoptosis, we cultured human dermal microvascular endothelial cells (HDMEC) with IFN-alpha at concentrations of 100, 500, 1,000 and 2,000 U/ml for 24-72 h and detected apoptosis by terminal deoxynucleotidyl transferase mediated FITC-dUTP nick end labeling (TUNEL). Quantitative analysis was performed using the FACScan and morphological alterations were studied by confocal laser scanning microscopy. In addition to the in vitro study we also analyzed frozen skin sections from proliferating, spontaneously regressing, and IFN-alpha-treated hemangiomas by simultaneous determination of endothelial cells and apoptotic nuclei using an indirect immunofluorescence test in combination with TUNEL.


RESULTS
Apoptosis was detected in up to 20% of the IFN-alpha-treated endothelial cells compared to the untreated controls. A maximum of apoptosis was observed after 48 h of stimulation with IFN-alpha in a dose-dependent manner. The analysis of hemangioma biopsies revealed apoptotic endothelial cells in IFN-alpha-treated as well as in spontaneously regressing hemangiomas, but not in proliferating ones.


CONCLUSIONS
These data indicate that the therapeutic effect of IFN-alpha on hemangiomas is based on apoptosis induction of endothelial cells, which might also explain the clinically and histologically observed involution without any sign of inflammation or necrosis.
Sgonc
R
R
Institute for General and Experimental Pathology, University of Innsbruck, Innsbruck, Austria. roswitha.sgone@uibk.ac.at
Fuerhapter
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C
Boeck
G
G
Swerlick
R
R
Fritsch
P
P
Sepp
N
N
D000906:Antibodies / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured / Q000187:drug effects / Q000502:physiology; D004396:Coloring Agents; D004247:DNA / Q000032:analysis; D053938:DNA Fragmentation; D020405:Dermis / Q000098:blood supply*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology; D005434:Flow Cytometry; D006391:Hemangioma / Q000188:drug therapy / Q000473:pathology* / Q000503:physiopathology*; D006801:Humans; D020287:In Situ Nick-End Labeling; D007223:Infant; D007231:Infant, Newborn; D016898:Interferon-alpha / Q000494:pharmacology* / Q000627:therapeutic use*; D008297:Male; D008833:Microcirculation; D018613:Microscopy, Confocal; D011419:Propidium; D016707:Tissue Fixation; D014841:von Willebrand Factor / Q000276:immunology
D000906:Antibodies; D004396:Coloring Agents; D016898:Interferon-alpha; D014841:von Willebrand Factor; D011419:Propidium; D004247:DNA