Cloning and functional expression of the murine homologue of proteinase 3: implications for the design of murine models of vasculitis

FEBS Letters

Hummel, Amber M.²  Jenne, Dieter E.¹  Fröhlich, Leopold¹  Specks, Ulrich² 

[1] Max-Planck-Institut für Psychiatrie, Abt. Neuroimmunologie, Martinsried, Germany;Thoracic Diseases Research Unit, Mayo Clinic and Foundation, Guggenheim Bldg. 642A, 200 First Street SW, Rochester, MN 55905, USA

关键词: Neutrophil;    Proteinase 3;    Anti-neutrophil cytoplasmic antibody;    In vivo animal model;    α1-PI;    α1-protease inhibitor (=α1-antitrypsin);    ANCA;    anti-neutrophil cytoplasmic antibodies;    c-ANCA;    cytoplasmic fluorescence pattern ANCA;    HNE;    human neutrophil elastase;    IIF;    indirect immunofluorescence;    mo-AB;    monoclonal antibody;    NGS;    normal goat serum;    PBS;    phosphate buffered saline;    PCR;    polymerase chain reaction;    PR3;    proteinase 3;    hPR3;    human PR3;    hPR3-tag;    human PR3 with His-Strep-tag;    mPR3;    mouse PR3;    mPR3-tag;    mouse PR3 with His-Strep-tag;    rPR3;    recombinant PR3;    TBS;    Tris buffered saline;    WG;    Wegener's granulomatosis;   

DOI  :  10.1016/S0014-5793(97)00418-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Anti-neutrophil cytoplasmic autoantibodies recognizing conformational epitopes (c-ANCA) of proteinase 3 (PR3) from azurophil granules are a diagnostic hallmark in Wegener's granulomatosis (WG). Because a functional PR3 homologue has not been identified in rodents, it is difficult to assess immunopathological responses in rats or mice immunized with patients' derived c-ANCA or human PR3. Here we report the full length cDNA cloning and functional expression of murine PR3 in HMC-1 cells. Recombinant murine PR3 shows highly similar substrate specificities towards synthetic peptides and is inhibited by human α1-proteinase inhibitor like human PR3. However, neither human c-ANCA, rabbit sera nor mouse monoclonal antibodies to human PR3 recognize the murine homologue. Consequently, it is unlikely that disease observed in mice after immunization with c-ANCA or human PR3 is caused by pathogenic antibodies directed against mouse PR3. Recombinant human-mouse chimaeric variants will be a valuable new tool to localize the disease-specific immunodominant epitopes in human PR3.

【 授权许可】

Unknown