Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis
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 ba...
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Wiley
2013-01-01
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| Series: | Clinical and Developmental Immunology |
| Online Access: | http://dx.doi.org/10.1155/2013/572815 |
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| author | Christiane Radzimski Christian Probst Bianca Teegen Kristin Rentzsch Inga Madeleine Blöcker Cornelia Dähnrich Wolfgang Schlumberger Winfried Stöcker Dimitrios P. Bogdanos Lars Komorowski |
| author_facet | Christiane Radzimski Christian Probst Bianca Teegen Kristin Rentzsch Inga Madeleine Blöcker Cornelia Dähnrich Wolfgang Schlumberger Winfried Stöcker Dimitrios P. Bogdanos Lars Komorowski |
| author_sort | Christiane Radzimski |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-8341a38e637446e29b1532125af2fe0d |
| institution | Kabale University |
| issn | 1740-2522 1740-2530 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Clinical and Developmental Immunology |
| spelling | doaj-art-8341a38e637446e29b1532125af2fe0d2025-02-03T05:47:43ZengWileyClinical and Developmental Immunology1740-25221740-25302013-01-01201310.1155/2013/572815572815Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune HepatitisChristiane Radzimski0Christian Probst1Bianca Teegen2Kristin Rentzsch3Inga Madeleine Blöcker4Cornelia Dähnrich5Wolfgang Schlumberger6Winfried Stöcker7Dimitrios P. Bogdanos8Lars Komorowski9Institute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyInstitute of Liver Studies, School of Medicine, King's College London, Denmark Hill, London SE5 9RS, UKInstitute of Experimental Immunology, Euroimmun AG, Seekamp 31, 23560 Lübeck, GermanyAutoantibodies 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.http://dx.doi.org/10.1155/2013/572815 |
| spellingShingle | Christiane Radzimski Christian Probst Bianca Teegen Kristin Rentzsch Inga Madeleine Blöcker Cornelia Dähnrich Wolfgang Schlumberger Winfried Stöcker Dimitrios P. Bogdanos Lars Komorowski Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis Clinical and Developmental Immunology |
| title | Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis |
| title_full | Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis |
| title_fullStr | Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis |
| title_full_unstemmed | Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis |
| title_short | Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis |
| title_sort | development of a recombinant cell based indirect immunofluorescence assay for the determination of autoantibodies against soluble liver antigen in autoimmune hepatitis |
| url | http://dx.doi.org/10.1155/2013/572815 |
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