Reference standards for the detection of anti-mitochondrial and anti-rods/rings autoantibodies

• Published in: Clinical chemistry and laboratory medicine Vol. 56; no. 10; pp. 1789 - 1798
• Main Authors: Calise, S. John, Zheng, Bing, Hasegawa, Tomoko, Satoh, Minoru, Isailovic, Natasa, Ceribelli, Angela, Andrade, Luis E.C., Boylan, Katherine, Cavazzana, Ilaria, Fritzler, Marvin J., de la Torre, Ignacio Garcia, Hiepe, Falk, Kohl, Kathryn, Selmi, Carlo, Shoenfeld, Yehuda, Tincani, Angela, Chan, Edward K.L.
• Format: Journal Article
• Language: English
• Published: Germany De Gruyter 25.09.2018; Walter De Gruyter & Company
• Subjects: Aged; anti-mitochondrial antibody; anti-rods/rings antibody; Antibodies; Antigens; Antiviral Agents - adverse effects; Antiviral Agents - therapeutic use; Autoantibodies; Autoantibodies - blood; Autoantibodies - immunology; autoantibody; Autoimmune Diseases - diagnosis; autoimmunity; Blood plasma; Cholangitis; Dehydrogenase; Dehydrogenases; Dilution; Enzyme-linked immunosorbent assay; Female; Fluorescent Antibody Technique, Indirect - standards; Hepatitis; Hepatitis C; Hepatitis C - drug therapy; Histones; Humans; Identification methods; Immunoassay; Immunoassay - methods; Immunoassay - standards; Immunofluorescence; Immunoprecipitation; Inosine monophosphate; Interferon; Interferon-alpha - adverse effects; Interferon-alpha - therapeutic use; IP (Internet Protocol); Kidneys; Laboratories; Liver; Middle Aged; Mitochondria; Mitochondria - immunology; Nucleosomes; Patients; primary biliary cholangitis; Proteins; Pyruvate dehydrogenase (lipoamide); Pyruvic acid; Reference materials; Reference Standards; Ribavirin; Ribavirin - adverse effects; Ribavirin - therapeutic use; Rods; Staining; Stomach; Test procedures; Western blotting; α-Interferon; anti-mitochondrial antibody; autoantibody; autoimmunity; hepatitis C; anti-rods/rings antibody; primary biliary cholangitis
• DOI: 10.1515/cclm-2017-1152
• ISSN: 1434-6621; 1437-4331

Abstract Background: Anti-mitochondrial antibodies (AMA) are found in >90% of primary biliary cholangitis patients. Anti-rods/rings antibodies (anti-RR) are most commonly associated with interferon-α and ribavirin treatment in hepatitis C patients. Clinical laboratories routinely screen for AMA and anti-RR using indirect immunofluorescence on HEp-2 cells (HEp-2-IFA). Therefore, we sought to establish reference materials for use in AMA and anti-RR testing. Methods: AMA-positive and anti-RR-positive human plasma samples (AMA-REF and RR-REF), identified as potential reference materials based on preliminary data, were further validated by multiple laboratories using HEp-2-IFA, immunoprecipitation (IP), western blotting, IP-western, line immunoassay (LIA), addressable laser bead immunoassay (ALBIA) and enzyme-linked immunosorbent assay (ELISA). Results: AMA-REF showed a strong positive cytoplasmic reticular/AMA staining pattern by HEp-2-IFA to ≥1:1280 dilution and positive signal on rodent kidney/stomach/liver tissue. AMA-REF reacted with E2/E3, E3BP, E1α and E1β subunits of the pyruvate dehydrogenase complex by IP and western blotting and was positive for AMA antigens by LIA, ALBIA and ELISA. RR-REF showed a strong positive rods and rings staining pattern by HEp-2-IFA to ≥1:1280 dilution. RR-REF reacted with inosine monophosphate dehydrogenase by IP, IP-western and ALBIA. RR-REF also produced a nuclear homogenous staining pattern by HEp-2-IFA, immunoprecipitated proteins associated with anti-U1RNP antibody and reacted weakly with histones, nucleosomes, Sm and nRNP/Sm by LIA. Conclusions: AMA-REF and RR-REF are useful reference materials for academic or commercial clinical laboratories to calibrate and establish internal reference standards for immunodiagnostic assays. AMA-REF and RR-REF are now available for free distribution to qualified laboratories through Plasma Services Group.