title: C4d Immunohistochemistry in glomerulonephritis with different antibodies.
abstract: BACKGROUND
  The presence of C4d in the kidney is generally detected particularly for the diagnosis of antibody-mediated rejection in renal transplants. In frozen sections of immunofluorescence (IF) staining with anti-C4d monoclonal antibodies (mAbs), we noted intrinsic C4d deposition even in normal glomeruli though their pathogenic or an intrinsic role is unkown. An anti-C4d polyclonal antibody (C4dpAb), which is suitable for paraffin immunoperoxidase (IP) staining, is less used than mAbs, and it has demonstrated that intrinsic C4d is not evident. To establish a stable and reproducible procedure for C4d detection with the C4dpAb and to determine the staining characteristics of it, the present study aimed to test whether the method was comparable with IF with a mAb.
  
  
  METHODS
  We compared the C4dpAb with the mAb in adjacent sections of human diseased kidneys, and then compared IP with IF of C4dpAb. Two ways of antigen retrieval was examined for IP.
  
  
  RESULTS
  On comparing the two antibodies for glomerular staining with IF, we found that the pattern and intensity (C4dpAb showed intrinsic C4d with IF) were similar. In addition, C4dpAb staining with IP and IF demonstrated that the intrinsic staining in the normal glomerulus was mostly undetectable by IP, whereas IF showed distinct staining. Likewise, C4d deposition with IP in some cases was apparently weaker than that on IF, suggesting that this deposition is not intrinsic but indicates pathogenic complement activation.
  
  
  CONCLUSIONS
  The advantage of the C4dpAb for immunohistochemistry is of value for reconsidering the role of C4d in glomerular diseases.
authors: Molecular Pathology, Biomolecular and Integrated Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennohdai, Tsukuba, Ibaraki, 305-8575, Japan.
authors: Taisei
authors: T
authors: Suzuki
authors: Kidney Center, Tokyo Women's Medical University, Tokyo, Japan.
authors: Shigeru
authors: S
authors: Horita
authors: The Pathology Department, Matsuyama Red Cross Hospital, Ehime, Japan.
authors: Koji
authors: K
authors: Kadoya
authors: The Kidney Center, Matsuyama Red Cross Hospital, Ehime, Japan.
authors: Koji
authors: K
authors: Mitsuiki
authors: Molecular Pathology, Biomolecular and Integrated Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennohdai, Tsukuba, Ibaraki, 305-8575, Japan.
authors: Kumi
authors: K
authors: Aita
authors: The Kidney Center, Matsuyama Red Cross Hospital, Ehime, Japan.
authors: Atsumi
authors: A
authors: Harada
authors: Kidney Center, Tokyo Women's Medical University, Tokyo, Japan.
authors: Kosaku
authors: K
authors: Nitta
authors: Molecular Pathology, Biomolecular and Integrated Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennohdai, Tsukuba, Ibaraki, 305-8575, Japan. nagatam@md.tsukuba.ac.jp.
authors: Michio
authors: M
authors: Nagata
mesh_terms: D000906:Antibodies; D000911:Antibodies, Monoclonal; D015935:Complement C4b / Q000032:analysis* / Q000276:immunology; D005455:Fluorescent Antibody Technique; D005629:Frozen Sections; D005921:Glomerulonephritis / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000276:immunology*; D016612:Paraffin Embedding; D010446:Peptide Fragments / Q000032:analysis* / Q000276:immunology; D015203:Reproducibility of Results
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D015935:Complement C4b; C032261:complement C4d