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Mytilinaiou M.G.,Kings College London | Mytilinaiou M.G.,Cell Immunotherapy and Molecular Immunodiagnostics Biomedical Research and Technology Biomedical | Meyer W.,Institute for Experimental Immunology affiliated to EUROIMMUN AG | Scheper T.,Institute for Experimental Immunology affiliated to EUROIMMUN AG | And 11 more authors.
Clinica Chimica Acta | Year: 2012

Background: The lack of an immunoassay that detects antibodies to promyelocytic leukaemia (PML) protein, the primary biliary cirrhosis (PBC)-specific multiple nuclear dot (MND) antigen, has prompted us to develop a line immunoassay (LIA) for the simultaneous detection of PML and Sp100 MND-specific autoantibodies. Methods: PML and Sp100 were expressed in Escherichia coli, and analysed by SDS-PAGE and immunoblotting using a monoclonal antibody and MALDI-ToF fingerprinting. A quantitative PML and Sp100 LIA were developed and testing was performed in 150 anti-mitochondrial antibody (AMA) positive, 20 AMA-PBCs and 130 controls. Results: Thirty-five (23%) of 150 AMA. + PBCs (18 anti-MND. +) were anti-PML. + (12%) or anti-Sp100. + (20%), 10 being anti-PML. +/Sp100+, 5 single anti-PML. + and 20 single anti-Sp100+. Six (30%, 5 anti-MND. +) AMA-PBCs were anti-PML. + or Sp100+. Only 2 (1.7%) pathological controls were anti-PML. + and/or anti-Sp100+. Levels of anti-PML correlated with those of anti-Sp100 (R = 0.64, p< 0.0001). The autoantibody profile largely remained unchanged over a 10. year-follow up (52 patients, 352 samples). Anti-PML, Sp100 or MND-reactive PBCs were younger and had longer disease duration than the seronegative (p = 0.06, for both). Anti-Sp100 levels correlated with the Mayo risk score (r = 0.63, p=0.01). Anti-PML. +/Sp100. + patients had more advanced disease compared to patients negative for anti-PML/Sp100 (p=0.04). Conclusion: The new line immunoassay offers a robust and accurate method for the detection of clinically-relevant PBC-specific anti-MND antibodies. © 2012 Elsevier B.V.

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