C4d Immunohistochemistry in glomerulonephritis with different antibodies.
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.
Suzuki
Taisei
T
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.
Horita
Shigeru
S
Kidney Center, Tokyo Women's Medical University, Tokyo, Japan.
Kadoya
Koji
K
The Pathology Department, Matsuyama Red Cross Hospital, Ehime, Japan.
Mitsuiki
Koji
K
The Kidney Center, Matsuyama Red Cross Hospital, Ehime, Japan.
Aita
Kumi
K
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.
Harada
Atsumi
A
The Kidney Center, Matsuyama Red Cross Hospital, Ehime, Japan.
Nitta
Kosaku
K
Kidney Center, Tokyo Women's Medical University, Tokyo, Japan.
Nagata
Michio
M
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.
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
D000906:Antibodies; D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D015935:Complement C4b; C032261:complement C4d