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-kappaB by measuring the loss of its inhibitor I-kappaB. 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-kappaB. 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-kappaB activation.
|Titolo:||Human monoclonal anti-endothelial cell IgG-derived from a systemic lupus erythematosus patient binds and activates human endothelium in vitro|
RASCHI, ELENA (Secondo)
MERONI, PIERLUIGI (Penultimo)
|Parole Chiave:||NF-kappa B; Vasculitis; Intercellular Adhesion Molecule-1; Autoimmune Diseases; Enzyme Activation; Humans; Transcription, Genetic; Endothelium, Vascular; Antibodies, Monoclonal; Antibody Specificity; Lupus Erythematosus, Systemic; Autoantibodies; Cells, Cultured; E-Selectin; JNK Mitogen-Activated Protein Kinases; I-kappa B Proteins; Immunoglobulin G; Interleukin-6; Mitogen-Activated Protein Kinases; Gene Expression Regulation; Umbilical Cord; U937 Cells; Cell Adhesion|
|Settore Scientifico Disciplinare:||Settore MED/16 - Reumatologia|
|Data di pubblicazione:||mar-2001|
|Appare nelle tipologie:||01 - Articolo su periodico|