title: Detection of antiendothelial cell antibodies by an enzyme-linked immunosorbent assay using antigens from cell lysate: minimal interference with antinuclear antibodies and rheumatoid factors.
abstract: The objective of the present work was to set up a routine test adapted to screening for antiendothelial cell antibodies (AECAs) in serum samples with minimal interference from antinuclear antibodies (ANAs) or rheumatoid factors (RFs). We compared the titers of AECAs titrated following two enzyme-linked immunosorbent assays (ELISAs): (i) an ELISA with ethanol-fixed EA.hy926 monolayers as the antigenic substrate and (ii) an ELISA with nucleus-depleted lysates prepared from EA.hy926 cells and normalized for protein (1.0 to 1.7 mg/ml) and DNA (< or =0.1 microg/ml) contents as a surrogate substrate (postnuclear supernatant ELISA [PNS-ELISA]). The AECA titers in 51 serum samples, including 28 samples containing ANAs, were compared. A significantly positive correlation (r = 0.77; P < 0.001) between the two series was shown only for the ANA-negative serum samples. Conversely, ANAs or RFs in samples were shown not to interfere in tests for AECAs by the PNS-ELISA. AECAs recognize their antigenic targets in postnuclear supernatants, which is representative of the endothelial antigenic content, with improvement of the reliability of the assay, a prerequisite to application of the assay for their evaluation in clinical practice.
authors.affiliation: Laboratoire d'Immunologie, Hôpital Sud, CHU Grenoble, Grenoble, France. Christian.Drouet@ujf-grenoble.fr
authors.forename: Christian
authors.initials: C
authors.lastname: Drouet
authors.forename: Marie-France
authors.initials: MF
authors.lastname: Nissou
authors.forename: Denise
authors.initials: D
authors.lastname: Ponard
authors.forename: Josiane
authors.initials: J
authors.lastname: Arvieux
authors.forename: Chantal
authors.initials: C
authors.lastname: Dumestre-Pérard
authors.forename: Philippe
authors.initials: P
authors.lastname: Gaudin
authors.forename: Bernard
authors.initials: B
authors.lastname: Imbert
authors.forename: Christian
authors.initials: C
authors.lastname: Massot
authors.forename: Françoise
authors.initials: F
authors.lastname: Sarrot-Reynauld
mesh_terms: D000974:Antibodies, Antinuclear / Q000097:blood / Q000276:immunology; D000941:Antigens / Q000276:immunology; D001323:Autoantibodies / Q000097:blood*; D001327:Autoimmune Diseases / Q000097:blood / Q000175:diagnosis*; D002460:Cell Line; D003429:Cross Reactions; D042783:Endothelial Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D015203:Reproducibility of Results; D012217:Rheumatoid Factor / Q000097:blood / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D000974:Antibodies, Antinuclear; D000941:Antigens; D001323:Autoantibodies; C087589:anti-endothelial cell antibody; D012217:Rheumatoid Factor