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Anti-endothelial cell (EC) antibodies (AECA) are a heterogeneous group of antibodies directed against a variety of EC membrane proteins. A pathogenic role for AECA in diseases that involve the vascular system has not been clearly demonstrated. Induction of EC apoptosis appears to be one of the mechanisms by which AECA may exert their effect. AECA from some patients trigger the translocation of anionic phospholipids, most notably phosphatidylserine, from the inner to the outer leaflet of the plasma membrane, and thereafter activation of caspase 3 and cleavage of poly (ADP-ribose) polymerase, hallmarks of apoptosis. Apoptotic cell death generates oxidatively modified moieties, which can induce autoimmune and local inflammatory responses. While a sole AECA target involved in the apoptotic process of ECs has not been identified, some evidence suggests that Heat Shock Proteins may be an outstanding antigen.

Heat-shock protein (Hsp) family is made up of heterogeneous proteins of which Hsp60 members are the most studied. It is now generally admitted that Hsp60 is not only a mitochondrial component but can be localized on the membrane cell surface. Considered as a signal danger following infections, Hsp60 can induce the production of anti-Hsp60 antibodies as defense mechanisms against pathogens. However, endogenous Hsp60 is also a target of autoantibodies in autoimmune disorders, atherosclerosis and vascular diseases, in which anti-endothelial cell antibodies (AECA) are generated. Hsp60 is one of the endothelial cell autoantigens able to trigger cytotoxic and apoptotic responses when recognized by the related autoantibodies. Depending on the Hsp60 epitope specificity, it appears that AECA with Hsp60 reactivity may differ in their functional effects. These observations suggest that new therapeutic approach to avoid endothelial cell damages due to anti-Hsp60 autoantibodies would be successful provided that specific Hsp60 epitopes would have been precisely characterized.

Objective: Anti-endothelial cell antibodies (AECAs), which recognize a number of endothelial antigens, are seen in patients with systemic autoimmune diseases, more often in the presence of vasculitis than in its absence. Some AECAs induce apoptosis of endothelial cells (ECs), but their target antigens remain unknown. The aim of this study was to determine whether Hsp60 is a target antigen and whether AECAs induce apoptosis in ECs., Methods: Two-dimensional electrophoresis and conventional Western blotting techniques were used to characterize AECA targets. Hsp60 reactivity was determined by enzyme-linked immunosorbent assay., Results: Hsp60 was shown to be targeted by a proportion of AECAs. The level of reactivity was higher in patients with systemic autoimmune disease and vasculitis than in those without vasculitis and in patients with systemic lupus erythematosus than in patients with other systemic autoimmune diseases. Hsp60 was expressed on the plasma membrane of heat-stressed ECs, and this followed Hsp60 messenger RNA transcription, confinement of the protein to the cytoplasm, and translocation of the protein to the surface. Shedding of Hsp60 from ECs was induced by stress and resulted in the binding of soluble Hsp60 to the surface of ECs, particularly stressed ECs. Apoptosis of ECs was triggered by anti-Hsp60-containing AECA-positive sera and was inhibited by preincubation of the ECs with recombinant Hsp60., Conclusion: Our data support the notion that Hsp60 is an important target for AECAs and that such an interaction contributes to pathogenic effects, especially in vasculitis-associated systemic autoimmune disease.