title: Development of a recombinant cell-based indirect immunofluorescence assay for the determination of autoantibodies against soluble liver antigen in autoimmune hepatitis.
abstract: Autoantibodies against soluble liver antigen (SLA) are specific markers for autoimmune hepatitis (AIH) type 1. In contrast to the determination of other AIH-associated autoantibodies by indirect immunofluorescence assay (IFA), detection of anti-SLA relied up to now on ELISA or immunoblot based on bacterially expressed recombinant protein. In order to develop a complementary IFA substrate, SLA isoform 1 was recombinantly produced in the human cell line HEK293 and controlled by a rabbit hyperimmune serum against SLA. The recombinant cells were used in IFA (RC-IFA) to analyze sera from 20 AIH patients with anti-SLA positivity predetermined by ELISA together with 80 controls (20 anti-SLA negative AIH, 15 primary biliary cirrhosis, 15 HCV, and 30 healthy blood donors). Using RC-IFA, anti-SLA was detected in all ELISA positive AIH sera but in none of the controls. Furthermore, a cytosolic fraction of HEK293 containing SLA was able to neutralize the autoantibodies in all positive sera in a dose-dependent manner. HEK293 cells expressing SLA are a valid substrate for the serodiagnosis of AIH relevant autoantibodies by IFA. In concert with cryosections of primate liver, rat kidney, rat liver, rat stomach, and HEp-2 cells, they enable the parallel determination of all autoantibodies associated with autoimmune liver diseases.
authors: Institute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, Germany.
authors: Christiane
authors: C
authors: Radzimski
authors: Christian
authors: C
authors: Probst
authors: Bianca
authors: B
authors: Teegen
authors: Kristin
authors: K
authors: Rentzsch
authors: Inga Madeleine
authors: IM
authors: Blöcker
authors: Cornelia
authors: C
authors: Dähnrich
authors: Wolfgang
authors: W
authors: Schlumberger
authors: Winfried
authors: W
authors: Stöcker
authors: Dimitrios P
authors: DP
authors: Bogdanos
authors: Lars
authors: L
authors: Komorowski
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000097:blood*; D001324:Autoantigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D003001:Cloning, Molecular; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D057809:HEK293 Cells; D019693:Hepatitis, Autoimmune / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D019076:Transgenes / Q000235:genetics
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D015415:Biomarkers; D020033:Protein Isoforms; C046431:liver antigen LA-1
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