Cell-based BSEP trans-inhibition: A novel, non-invasive test for diagnosis of antibody-induced BSEP deficiency.

Autor:	Stindt J; Department of Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany., Dröge C; Department of Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.; Department of Gastroenterology, Hepatology and Infectious Diseases, University Hospital Magdeburg, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany., Lainka E; Department of Pediatrics, Division of Gastroenterology and Hepatology, University Children's Hospital, Essen, Germany., Kathemann S; Department of Pediatrics, Division of Gastroenterology and Hepatology, University Children's Hospital, Essen, Germany., Pfister ED; Pediatric Gastroenterology and Hepatology, Department for Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, Hannover, Germany., Baumann U; Pediatric Gastroenterology and Hepatology, Department for Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, Hannover, Germany., Stalke A; Pediatric Gastroenterology and Hepatology, Department for Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, Hannover, Germany.; Department of Human Genetics, Hannover Medical School, Hannover, Germany., Grabhorn E; Pediatric Hepatology and Liver Transplantation, Transplantation Center, University Medical Center Hamburg-Eppendorf, Hamburg, Germany., Shagrani MA; Department of Liver & SB Transplant & Hepatobiliary-Pancreatic Surgery, King Faisal Specialist Hospital & Research Centre, Riyadh, Saudi Arabia., Mozer-Glassberg Y; Institute of Gastroenterology, Nutrition and Liver Diseases, Schneider Children's Medical Centre of Israel, Petach Tikvah, Israel., Hartley J; The Liver Unit Including Small Bowel Transplantation, Birmingham Women's and Children's Hospital, Birmingham, UK., Wammers M; Department of Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany., Klindt C; Department of Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany., Philippski P; Department of Gastroenterology, Hepatology and Infectious Diseases, University Hospital Magdeburg, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany., Liebe R; Department of Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany., Herebian D; Department of General Pediatrics, Neonatology and Pediatric Cardiology, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany., Mayatepek E; Department of General Pediatrics, Neonatology and Pediatric Cardiology, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany., Berg T; Division of Hepatology, Department of Medicine II, Leipzig University Medical Center, Leipzig, Germany., Schmidt-Choudhury A; Department of Pediatrics and Adolescent Medicine, Ruhr University Bochum, Bochum, Germany., Wiek C; Department of Otorhinolaryngology, Heinrich Heine University School of Medicine, Düsseldorf, Germany., Hanenberg H; Department of Otorhinolaryngology, Heinrich Heine University School of Medicine, Düsseldorf, Germany.; Department of Pediatrics III, University Children's Hospital, University of Duisburg-Essen, Essen, Germany., Luedde T; Department of Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany., Keitel V; Department of Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.; Department of Gastroenterology, Hepatology and Infectious Diseases, University Hospital Magdeburg, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany.
Jazyk:	angličtina
Zdroj:	JHEP reports : innovation in hepatology [JHEP Rep] 2023 Feb 01; Vol. 5 (7), pp. 100690. Date of Electronic Publication: 2023 Feb 01 (Print Publication: 2023).
DOI:	10.1016/j.jhepr.2023.100690
Abstrakt:	Background & Aims: Antibody-induced bile salt export pump deficiency (AIBD) is an acquired form of intrahepatic cholestasis, which may develop following orthotopic liver transplantation (OLT) for progressive familial intrahepatic cholestasis type 2 (PFIC-2). Approximately 8-33% of patients with PFIC-2 who underwent a transplant develop bile salt export pump (BSEP) antibodies, which trans-inhibit this bile salt transporter from the extracellular, biliary side. AIBD is diagnosed by demonstration of BSEP-reactive and BSEP-inhibitory antibodies in patient serum. We developed a cell-based test directly measuring BSEP trans-inhibition by antibodies in serum samples to confirm AIBD diagnosis. Methods: Sera from healthy controls and cholestatic non-AIBD or AIBD cases were tested (1) for anticanalicular reactivity by immunofluorescence staining of human liver cryosections, (2) for anti-BSEP reactivity by immunofluorescence staining of human embryonic kidney 293 (HEK293) cells expressing BSEP-enhanced yellow fluorescent protein (EYFP) and immunodetection of BSEP-EYFP on Western blot, and (3) for BSEP trans-inhibition using HEK293 cells stably expressing Na + /taurocholate cotransporting polypeptide (NTCP)-mCherry and BSEP-EYFP. The trans-inhibition test uses [ 3 H]-taurocholate as substrate and is divided into an uptake phase dominated by NTCP followed by BSEP-mediated export. For functional analysis, sera were bile salt depleted. Results: We found BSEP trans-inhibition by seven sera containing anti-BSEP antibodies, but not by five cholestatic or nine control sera, all lacking BSEP reactivity. Prospective screening of a patient with PFIC-2 post OLT showed seroconversion to AIBD, and the novel test method allowed monitoring of treatment response. Notably, we identified a patient with PFIC-2 post OLT with anti-BSEP antibodies yet without BSEP trans-inhibition activity, in line with asymptomatic presentation at serum sampling. Conclusions: Our cell-based assay is the first direct functional test for AIBD and allows confirmation of diagnosis as well as monitoring under therapy. We propose an updated workflow for AIBD diagnosis including this functional assay. Impact and Implications: Antibody-induced BSEP deficiency (AIBD) is a potentially serious complication that may affect patients with PFIC-2 after liver transplantation. To improve its early diagnosis and thus immediate treatment, we developed a novel functional assay to confirm AIBD diagnosis using a patient's serum and propose an updated diagnostic algorithm for AIBD. Competing Interests: EL has received honoraria for lectures from Albireo and Mirum and has served on advisory boards for both companies and on the GPGE board of directors. UB has received grants from Mirum Pharma, Albireo Pharma, and Alexion Pharma and has received consulting fees and honoraria from Mirum, Albireo, Alexion, Nestle, and Vivet. CK has received travel grant from Falk. TB has received grants, consulting fees, and/or honoraria from Abbvie, Alexion, Bayer, BMS, Gilead, GSK, Eisai, Enyo Pharma, Falk Foundation, HepaRegeniX GmbH, Humedics, Intercept, Ipsen, Janssen, Medupdate GmbH, MSD/Merck, Merz, Norgine, Novartis, Orphalan, Roche, Sequana Medical, SIRTEX, SOBI, and Shionogi. VK has received honoraria for speaker’s bureau from Falk, Albireo, CSL, Sanofi, Gilead, Abbvie, and Intercept and has served on one advisory board for Astra Zeneca. All other authors declare that there is no potential conflict of interest. Please refer to the accompanying ICMJE disclosure forms for further details. (© 2023 The Authors.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu