Working Algorithms and Detection Methods of Autoantibodies in Autoimmune Liver Disease: A Nationwide Study

Autoantibody detection is the cornerstone of autoimmune liver diseases (AILD) diagnosis. Standardisation of working algorithms among autoimmunity laboratories, as well as being aware of the sensitivity and specificity of various commercial techniques in daily practice, are still necessary. The aim of this nationwide study is to report the results of the 2020 Autoimmunity Workshop organised by the Autoimmunity Group of the Spanish Society of Immunology and to provide useful information to clinicians and laboratory specialists to improve the management of autoantibody detection in AILD diagnoses. Serum samples from 17 patients with liver diseases were provided by the organisers of the 2020 Autoimmunity Workshop and were subsequently analysed by the 40 participating laboratories. Each laboratory used different techniques for the detection of autoantibodies in each patients’ serum sample, according to their working algorithm. Thus, almost 680 total complete patient reports were obtained, and the number of results from different autoantibody detection techniques was >3000. Up to eight different working algorithms were employed, including indirect immunofluorescence assays (IFA) and antigen-specific techniques (AgST). The IFA of HEp-2 cells was more sensitive than IFA of rat triple tissue for the study of anti-nuclear autoantibodies (ANA) associated with AILD. The IFA of a human neutrophil study for the analysis of anti-neutrophil cytoplasmic autoantibodies was not carried out systemically in all patients, or by all laboratories. AgSTs were the most sensitive methods for the detection of anti-smooth muscle/F-actin, soluble liver antigen, liver cytosol-1, M2-mitochondrial autoantibodies, and ANA associated with primary biliary cholangitis. The main differences in AMA detection were due to patients with autoantibodies against the non-dominant epitope of pyruvate dehydrogenase complex. Given that they are complementary, IFA and AgST should be performed in parallel. If there is high suspicion of AILD, AgST should always be performed.

[1]  K. Tsuneyama,et al.  Clinical practice guidelines for autoimmune hepatitis , 2022, Hepatology research : the official journal of the Japan Society of Hepatology.

[2]  J. Climent,et al.  Detection of anti-mitochondrial 2-oxoacid dehydrogenase complex subunit's antibodies for the diagnosis of primary biliary cholangitis. , 2021, Clinical immunology.

[3]  Linda Beenet.  The major diagnostic role of autoantibodies in the diagnosis of autoimmune hepatitis, a disease of all ages , 2021, Clinical and molecular hepatology.

[4]  F. Carrilho,et al.  Antineutrophil cytoplasmic antibody profiles differ according to type of primary sclerosing cholangitis and autoimmune hepatitis , 2021, Clinics.

[5]  A. Tanaka.  Current understanding of primary biliary cholangitis , 2020, Clinical and molecular hepatology.

[6]  A. Franke,et al.  Anti‐glycoprotein 2 (anti‐GP2) IgA and anti‐neutrophil cytoplasmic antibodies to serine proteinase 3 (PR3‐ANCA): antibodies to predict severe disease, poor survival and cholangiocarcinoma in primary sclerosing cholangitis , 2020, Alimentary pharmacology & therapeutics.

[7]  A. Lohse,et al.  Update of the simplified criteria for autoimmune hepatitis: evaluation of the methodology for immunoserological testing. , 2020, Journal of hepatology.

[8]  M. Mahler,et al.  Novel Anti-Hexokinase 1 Antibodies Are Associated With Poor Prognosis in Patients With Primary Biliary Cholangitis. , 2020, The American journal of gastroenterology.

[9]  Lisa K. Peterson,et al.  Assessment of antinuclear antibodies by indirect immunofluorescence assay: report from a survey by the American Association of Medical Laboratory Immunologists , 2020, Clinical chemistry and laboratory medicine.

[10]  D. Thorburn,et al.  Guideline review: British Society of Gastroenterology/UK-PSC guidelines for the diagnosis and management of primary sclerosing cholangitis , 2020, Frontline Gastroenterology.

[11]  M. Manns,et al.  Diagnosis and Management of Autoimmune Hepatitis in Adults and Children: 2019 Practice Guidance and Guidelines From the American Association for the Study of Liver Diseases , 2019, Hepatology.

[12]  C. Mack,et al.  The Contribution of B Cells in Autoimmune Liver Diseases , 2019, Seminars in Liver Disease.

[13]  M. Fritzler,et al.  The Prevalence of Anti-Hexokinase-1 and Anti-Kelch-Like 12 Peptide Antibodies in Patients With Primary Biliary Cholangitis Is Similar in Europe and North America: A Large International, Multi-Center Study , 2019, Front. Immunol..

[14]  J. Koskinas,et al.  Hellenic Association for the Study of the Liver Clinical Practice Guidelines: Autoimmune hepatitis , 2018, Annals of gastroenterology.

[15]  W. Kim,et al.  Diagnosis and Management of Primary Biliary Cholangitis , 2019, The American Journal of Gastroenterology.

[16]  T. Scheper,et al.  Anti-Ro60 Seropositivity Determines Anti-Ro52 Epitope Mapping in Patients With Systemic Sclerosis , 2018, Front. Immunol..

[17]  M. Manns,et al.  Autoimmune hepatitis , 2018, Nature Reviews Disease Primers.

[18]  T. Liwinski,et al.  Autoantibodies in Autoimmune Liver Disease—Clinical and Diagnostic Relevance , 2018, Front. Immunol..

[19]  U. Christen,et al.  Autoantibodies in Autoimmune Hepatitis: Can Epitopes Tell Us about the Etiology of the Disease? , 2018, Front. Immunol..

[20]  A. Parés.  Primary biliary cholangitis. , 2018, Medicina clinica.

[21]  J. Tabibian,et al.  Primary sclerosing cholangitis: A review and update. , 2017, Liver research.

[22]  G. Hirschfield,et al.  EASL Clinical Practice Guidelines: The diagnosis and management of patients with primary biliary cholangitis. , 2017, Journal of hepatology.

[23]  M. Gershwin,et al.  A contemporary perspective on the molecular characteristics of mitochondrial autoantigens and diagnosis in primary biliary cholangitis , 2016, Expert review of molecular diagnostics.

[24]  Paulo Luiz Carvalho Francescantônio,et al.  Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp-2 Cell Patterns 2014–2015 , 2015, Front. Immunol..

[25]  D. Bogdanos,et al.  Anti‐SLA/LP alone or in combination with anti‐Ro52 and fine specificity of anti‐Ro52 antibodies in patients with autoimmune hepatitis , 2015, Liver international : official journal of the International Association for the Study of the Liver.

[26]  J. Drenth.  EASL Clinical Practice Guidelines: Autoimmune hepatitis. , 2015, Journal of hepatology.

[27]  Shiling Hu,et al.  The accuracy of the anti-mitochondrial antibody and the M2 subtype test for diagnosis of primary biliary cirrhosis: a meta-analysis , 2014, Clinical chemistry and laboratory medicine.

[28]  Minoru Nakamura.  Clinical Significance of Autoantibodies in Primary Biliary Cirrhosis , 2014, Seminars in Liver Disease.

[29]  Q. Hu,et al.  Meta-Analysis Assessment of GP210 and SP100 for the Diagnosis of Primary Biliary Cirrhosis , 2014, PloS one.

[30]  Torsten Witte,et al.  International recommendations for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies , 2013, Annals of the rheumatic diseases.

[31]  D. Vergani,et al.  Clinical significance of autoantibodies in autoimmune hepatitis. , 2013, Journal of autoimmunity.

[32]  C. V. van Nieuwkerk,et al.  Auto immune hepatitis. , 2016, World journal of gastroenterology.

[33]  E. Ballot,et al.  Auto-antibodies in autoimmune hepatitis: anti-smooth muscle antibodies (ASMA). , 2012, Clinics and research in hepatology and gastroenterology.

[34]  H. Tony,et al.  Nuclear Pore Protein p62 Autoantibodies in Systemic Lupus Erythematosus , 2010, The open rheumatology journal.

[35]  T. Sauerbruch,et al.  p-ANCAs in autoimmune liver disorders recognise human β-tubulin isotype 5 and cross-react with microbial protein FtsZ , 2009, Gut.

[36]  I. Mackay,et al.  Autoimmune liver serology: current diagnostic and clinical challenges. , 2008, World journal of gastroenterology.

[37]  R. Chapman,et al.  Antibodies to SS‐A/Ro‐52kD and centromere in autoimmune liver disease: a clue to diagnosis and prognosis of primary biliary cirrhosis , 2007, Alimentary pharmacology & therapeutics.

[38]  J. Węsierska‐Gądek,et al.  Characterization of Autoantibodies against Components of the Nuclear Pore Complexes , 2007, Annals of the New York Academy of Sciences.

[39]  D. Bogdanos,et al.  Diagnostic relevance of anti-filamentous actin antibodies in autoimmune hepatitis , 2007, Journal of Clinical Pathology.

[40]  Maryland.,et al.  Manual of Molecular and Clinical Laboratory Immunology , 2006 .

[41]  M. Manns,et al.  Liver autoimmune serology: a consensus statement from the committee for autoimmune serology of the International Autoimmune Hepatitis Group. , 2004, Journal of hepatology.

[42]  J. B. Rattner,et al.  The nuclear pore complex protein Tpr is a common autoantigen in sera that demonstrate nuclear envelope staining by indirect immunofluorescence , 2004, Clinical and experimental immunology.

[43]  F. Alvarez,et al.  Anti-Soluble Liver Antigen (SLA) Antibodies in Chronic HCV Infection , 2004, Autoimmunity.

[44]  J. Sánchez-Román,et al.  Clinical significance of anti-multiple nuclear dots/Sp100 autoantibodies. , 2003, Scandinavian journal of gastroenterology.

[45]  M. Manns,et al.  Prävalenz von zirkulierenden Autoantikörpern bei gesunden Individuen , 2002, Medizinische Klinik.

[46]  J. Herkel,et al.  Identification of target antigen for SLA/LP autoantibodies in autoimmune hepatitis , 2000, The Lancet.

[47]  M. Manns,et al.  International Autoimmune Hepatitis Group Report: review of criteria for diagnosis of autoimmune hepatitis. , 1999, Journal of hepatology.

[48]  F. Alvarez,et al.  Formiminotransferase cyclodeaminase is an organ-specific autoantigen recognized by sera of patients with autoimmune hepatitis. , 1999, Gastroenterology.

[49]  M. Lenzi,et al.  Liver/kidney microsomal antibody type 1 and liver cytosol antibody type 1 concentrations in type 2 autoimmune hepatitis , 1998, Gut.

[50]  K. Boberg,et al.  Incidence and prevalence of primary biliary cirrhosis, primary sclerosing cholangitis, and autoimmune hepatitis in a Norwegian population. , 1998, Scandinavian journal of gastroenterology.

[51]  J. Koziol,et al.  Range of antinuclear antibodies in "healthy" individuals. , 1997, Arthritis and rheumatism.

[52]  D. Vergani,et al.  Autoimmune hepatitis in childhood: A 20‐year experience , 1997, Hepatology.

[53]  A. Czaja,et al.  Frequency and significance of antibodies to actin in type 1 autoimmune hepatitis , 1996, Hepatology.

[54]  M. Lenzi,et al.  Liver cytosolic 1 antigen-antibody system in type 2 autoimmune hepatitis and hepatitis C virus infection. , 1995, Gut.

[55]  G. Blobel,et al.  Identification and characterization of autoantibodies against the nuclear envelope lamin B receptor from patients with primary biliary cirrhosis , 1990, The Journal of experimental medicine.

[56]  C. Johanet,et al.  Antibody to liver cytosol (anti‐LC1) in patients with autoimmune chronic active hepatitis type 2 , 1988, Hepatology.

[57]  F. Alvarez,et al.  Chronic active hepatitis associated with antiliver/kidney microsome antibody type 1: A second type of “autoimmune” hepatitis , 1987, Hepatology.

[58]  D. Doniach,et al.  Classification of smooth muscle autoantibodies detected by immunofluorescence. , 1976, Journal of clinical pathology.

[59]  E. Holborow,et al.  Antibody to smooth muscle in patients with liver disease. , 1965, Lancet.

[60]  J. Sans-Sabrafen,et al.  [Lupoid hepatitis]. , 1965, Medicina clinica.