International Immunology, Vol. 13, No. 3, 349-357,
March 2001
© 2001 Japanese Society for Immunology
Human monoclonal anti-endothelial cell IgG-derived from a systemic lupus erythematosus patient binds and activates human endothelium in vitro
Department of Biomedical Sciences, University of Bradford, Bradford BD7 1DP, UK
1 Department of Internal Medicine, IRCCS Ospedale Maggiore, Via Sforza 35, 20145 Milan, Italy
2 Department of Internal Medicine, IRCCS Istituto Auxologico Italiano, University of Milan, Via L. Ariosto 13, 20145 Milan, Italy.
Correspondence to: P. L. Meroni
Our objectives were to obtain monoclonal anti-endothelial cell antibodies (AECA) from systemic lupus erythematosus (SLE) patients, to characterize their antigen specificity, and their capability to induce a pro-inflammatory and pro-adhesive endothelial phenotype, and to investigate the mechanism of endothelial cell (EC) activation in vitro. Monoclonal IgG AECA were generated by hybridoma formation with human SLE B cells. Antigen specificity was characterized by immunoblotting with enriched cell membrane fractions, by cytofluorimetry and by cell solid-phase ELISA. Endothelial activation was evaluated by measuring increases in U937 cell adhesiveness, adhesion molecule (E-selectin and ICAM-1) expression and IL-6 production. In addition, mechanisms of endothelial activation were investigated by assessment of NF-
B by measuring the loss of its inhibitor I-B. mAb E-3 bound live EC and recognized a 42 kDa EC membrane protein, it enhanced U937 adhesiveness, E-selectin and ICAM-1 expression and IL-6 production, and caused the loss of I-B. We conclude this is the first in vitro demonstration that a human monoclonal AECA from a SLE patient reacts with a constitutive endothelial membrane antigen and induces a pro-inflammatory endothelial phenotype through NF-B activation.
Keywords: adhesion molecules, anti-endothelial cell antibody, cytokines, systemic lupus erythematosus
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