ESPN 51th Annual Meeting

ESPN 2018


 
IMPAIRED ENDOTHELIAL CELL MIGRATION: NEW INSIGHTS INTO THE MECHANISMS OF COMPLEMENT-MEDIATED ENDOTHELIAL CELL INJURY
VALENTINA BRUNO 1 MAGDALENA RIEDL 2 CHIA WEI TEOH 2 CARMINE PECORARO 1 CHRISTOPH LICHT 2

1- S.C. NEFROLOGIA E DIALISI - SANTOBONO-PAUSILIPON CHILDREN’S HOSPITAL, NAPLES, ITALY
2- THE HOSPITAL FOR SICK CHILDREN, UNIVERSITY OF TORONTO, TORONTO, CANADA
 
Introduction:

Recent evidence suggests impairment of cell migration as a new mechanism of complement-mediated endothelial cell (EC) injury leading to thrombotic microangiopathy (TMA). We aimed to evaluate whether complement activation affects EC migration. 

Material and methods:

Healthy donor blood outgrowth endothelial cells (BOECs) were sensitized with anti-CD59 antibody and exposed to 50% Normal Human Serum (50% NHS). As controls, BOECs were exposed to 50% NHS without sensitization with anti-CD59 or to 50% Heat Inactivated Serum (50% HIS, complement inactive). A cell viability/proliferation assay was performed to evaluate the percentage of necrosis/apoptosis and proliferation. A scratch wound assay was used to study cell migration

Results:

We found signicant dierences (p<0.001) in wound closure between BOECs exposed to complement or not. Compared to controls, wound closure was reduced in BOECs after 4, 6, 8 and 10 hours of complement activation (11.59% vs 22.95% after 4 hours; 19.99% vs 38.27% after 6 hours; 31.99% vs 53.03% after 8 hours; 43.20% vs 67.43% after 10 hours. Figure). Cell death was excluded as cause for the defective wound closure (no significant difference in the rate of proliferation/viability compared to controls). We also found that impairment of BOEC migration was reversible after a short complement exposure (2-4 hours).

 

Conclusions:

We described impaired cell migration as a new mechanism of complement-mediated EC injury.