Epitope mapping of anti‐PR3 antibodies using chimeric human/mouse PR3 recombinant proteins

• Published in: Clinical and experimental immunology Vol. 135; no. 1; pp. 164 - 172
• Main Authors: SELGA, D., SEGELMARK, M., WIESLANDER, J., GUNNARSSON, L., HELLMARK, T.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.01.2004; Blackwell; Oxford University Press; Blackwell Science Inc
• Subjects: Amino Acid Sequence; ANCA; Animals; Antibodies, Antineutrophil Cytoplasmic - blood; Antibodies, Antineutrophil Cytoplasmic - immunology; Antibodies, Monoclonal - immunology; Autoantibodies - immunology; Basic Medicine; Biological and medical sciences; Cells, Cultured; Clinical Studies; Enzyme-Linked Immunosorbent Assay - methods; epitope; Epitope Mapping - methods; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Genetic Vectors; Granulomatosis with Polyangiitis - diagnosis; Granulomatosis with Polyangiitis - immunology; Humans; Immunologi inom det medicinska området; Immunology in the medical area; Immunopathology; Leukocyte Elastase - genetics; Medical and Health Sciences; Medical sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Mice; Molecular Sequence Data; Myeloblastin; Peroxidase - immunology; proteinase 3; Recombinant Fusion Proteins - immunology; Recombinant Proteins - immunology; Serine Endopeptidases - genetics; Serine Endopeptidases - immunology; Transfection; vasculitis; Wegener's granulomatosis; Human; Cartography; Immunopathology; Serine endopeptidases; Antigenic determinant; Enzyme; Antibody; Rodentia; Peptidases; Vertebrata; Mammalia; Immunology; Mouse; Animal; ANCA proteinase 3 epitope vasculitis Wegener's granulomatosis; Hydrolases; Chimerism; Recombinant protein; Myeloblastin
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1111/j.1365-2249.2004.02314.x

SUMMARY Autoantibodies against proteinase 3 (PR3) and myeloperoxidase (MPO) (ANCA = anti‐neutrophil cytoplasmic antibodies) are used as diagnostic tools for patients with small vessel vasculitis. ANCA are detected by different assays, but the correlation between the results of these assays is generally poor. The overall aim of the study was to provide a framework for the future development of new assays with an increased diagnostic yield. In order to express discrete epitopes of human PR3 (hPR3), the nonantigenic molecules murine PR3 (mPR3) and human leucocyte elastase (HLE) were used as a framework. We constructed recombinant chimeric vectors and were able to produce 6 hPR3/mPR3 proteins and 3 hPR3/HLE proteins. Anti‐PR3 monoclonal antibodies differed in their binding pattern to the chimeras, but no distinct binding region could be identified for any monoclonal antibody. The recombinant hPR3/mPR3 were also tested in ELISA with sera from patients with Wegener's granulomatosis with renal involvement. The results show that patients have antibodies to different constructs, indicating that the patients vary in their antibody repertoire from the beginning of the disease, and that patients may have antibodies from a broad range of clones early in the course of the disease. Recombinant hPR3/mPR3 chimeric proteins have a potential to be used as antigens in future ANCA assays.