Anti-PR-3 antibodies induce endothelial IL-8 release.
Eur J Clin Invest
; 29(11): 973-9, 1999 Nov.
Article
in En
| MEDLINE
| ID: mdl-10583443
ABSTRACT
BACKGROUND:
It has been shown that interaction of anti-PR-3 antibodies with human endothelial cells (EC) leads to an activation of EC in vitro, i.e. induction of adhesion molecules like E-selectin, VCAM-1 and tissue factor. The aim of this study was to investigate the effect of anti-PR-3 antibodies on endothelial IL-8 expression. MATERIALS ANDMETHODS:
EC were cultured in 96-well plates and stimulated with TNF-alpha and IL-1beta for 1 h to induce membrane expression of endothelial PR-3. Anti-PR-3 antibodies were purified from sera from patients with clinically active Wegener's granulomatosis. Purified anti-Ro, anti-centromere, anti-dsDNA antibodies and a monoclonal anti-PR-3 antibody (WGM2) served as controls. Induction of IL-8 mRNA was detected by RT-PCR. IL-8 was measured in the supernatant of EC by ELISA. In addition, induction of NFkappaB was investigated by PAGE of nuclear extracts of EC and Western blot with ab against p65.RESULTS:
In contrast to controls, interaction of anti-PR-3 antibodies (patients and WGM2) with cytokine activated EC led to the highest amount of IL-8 synthesis. Priming of EC with cytokines alone induced a markedly lower IL-8 level. The lowest levels of IL-8 could be measured after incubation of unprimed EC with anti-PR-3 antibodies. Anti-PR-3 antibody induced endothelial IL-8 expression could be inhibited by cycloheximide. In addition, we established that the activation of NF-kappaB is critically involved in anti-PR-3 antibody induced endothelial IL-8 production.CONCLUSION:
In summary, we were able to show that anti-PR-3 antibodies induce endothelial IL-8 synthesis by activating NF-kappaB. As IL-8 represents a powerful neutrophil chemoattractant, our data provide further evidence for a direct pathogenic effect of anti-PR-3 antibodies in ANCA related diseases.
Search on Google
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Autoantibodies
/
Autoantigens
/
Transcription, Genetic
/
Endothelium, Vascular
/
Serine Endopeptidases
/
Granulomatosis with Polyangiitis
/
Interleukin-8
Limits:
Humans
Language:
En
Journal:
Eur J Clin Invest
Year:
1999
Document type:
Article
Affiliation country:
Germany