Antibody determination by ELISA in rats with retinal S antigen-induced uveoretinitis

Abstract

Enzyme-linked immunosorbent assay was applied to the determination of the serum IgG antibody contents in rats immunized with the organ-specific autoantigen (S antigen) of the retina. Optimal conditions (i.e. S antigen concentration, serum and conjugate dilutions, enzymatic reaction time) were determined. The assay required only a single serum dilution and was well reproducible. It was very sensitive, allowing the detection of low antibody contents in animals immunized with 1 microgram of S antigen. The time course of the antibody response and its variations according to the dose and the species of the immunizing S antigen were considered.

Development of Monoclonal Antibodies to the Malondialdehyde−Deoxyguanosine Adduct, Pyrimidopurinone1

“…After the wells were washed four times with PBS-T, they were incubated for 1 h with aliquots of diluted peroxidase-labeled goat anti-mouse immunoglobulin (Chemicon). After washing, peroxidase activity remaining in the wells was determined by adding 0.05 M sodium phosphate/0.024 M citrate buffer (pH 5.0), containing 0.04% o-phenylenediamine and 0.012% hydrogen peroxide (200 µL/well) (29). The reaction was stopped after 5 min with 8 N sulfuric acid (50 µL/well), and the absorbance was measured at 490 nm using a Titertek Multiscan microtiter plate reader.…”

Rétine et auto-immunité

“…L'immunisation avec l' Ag-S, l'IRBP ou la rétine totale provoque une pinéalite associée à 1 'uvéa-rétinite. 1 La réponse immunitaire à l'antigène-S Outre l'atteinte pathologique de l'oeil , la réponse immune com porte la production d'anticorps circulants, qui sont déterminés habituellement aujourd'hui par ELISA [12], et une réponse im mune cellulaire. Celle-ci peut être mesurée par les tests de lympho prolifération ou de production de lymphokines (inhibition de migra tion des macrophages) induites in vitro par l'auto-antigène.…”