A sensitive immunoenzymometric assay for 2',5'-oligoadenylate. Detection of elevated 2',5'-oligoadenylate synthetase in human peripheral mononuclear cells.
A competition immunoenzymometric assay for 2',5'-oligoadenylate was developed and employed to measure the interferon-inducible enzyme 2',5'-oligoadenylate synthetase in cell extracts. Microtiter plates coated with a novel conjugate of p5'A2'p5'A2'p5'A and N-(2-aminoethyl)-carbamylmethylated-Ficoll (AECM-Ficoll) bound rabbit polyclonal or mouse monoclonal antibody directed against 2',5'-oligoadenylate. Binding was inhibited by soluble 2',5'-oligoadenylate. Estimates of 2',5'-oligoadenylate concentrations based on inhibition of antibody binding compared favorably with those obtained using a protein synthesis inhibition assay. Low concentrations of 2',5'-oligoadenylate synthesized in vitro by extracts of human peripheral mononuclear cells were conveniently estimated using less than or equal to 10(6) cells. Virtually identical results were obtained when either total extract or synthetase bound to poly(I) . poly(C)-agarose was used for the in vitro incubation. When peripheral mononuclear cells were incubated in vitro with interferon, the normally low levels of 2',5'-oligo(A) synthetase rose dramatically. The assay was employed to measure synthetase levels in peripheral mononuclear cells isolated from patients with systemic lupus erythematosus. Some of these patients were found to have elevated levels of 2',5'-oligoadenylate synthetase.
D015088:2',5'-Oligoadenylate Synthetase / Q000096:biosynthesis / Q000097:blood* / Q000276:immunology; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation; D006801:Humans; D007124:Immunoenzyme Techniques; D007370:Interferon Type I / Q000502:physiology; D007962:Leukocytes / Q000201:enzymology* / Q000276:immunology; D008180:Lupus Erythematosus, Systemic / Q000201:enzymology / Q000276:immunology; D011070:Poly I-C
D007370:Interferon Type I; D015088:2',5'-Oligoadenylate Synthetase; D011070:Poly I-C