Preliminary technical report. Epitope-defined monoclonal antibodies against type-IV collagen for diagnosis of Alport's syndrome
| Autor: | Yoshifumi Ninomiya, Tadashi Nemoto, Megumi Kagawa, Yumiko Kishiro, Yoshikazu Sado, Hiroshi Nakanishi, Ichiro Naito |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Transplantation
Pathology medicine.medical_specialty biology business.industry medicine.drug_class Gene mutation Monoclonal antibody medicine.disease Primary and secondary antibodies Molecular biology Epitope Staining Type IV collagen Nephrology biology.protein Medicine Alport syndrome Antibody business |
| Zdroj: | Nephrology Dialysis Transplantation. 12:1238-1241 |
| ISSN: | 1460-2385 0931-0509 |
| Popis: | chains of type-IV collagen [1‐3 ]. The genes of the Background. Alport’s syndrome can be diagnosed by a3( IV ) and a4 (IV ) chains are on chromosome 2 in a staining the a5 chain of type IV collagen in kidney head-to-head fashion [4 ], whereas the gene for the biopsy specimens with a monoclonal antibody. Because a5( IV ) chain resides on chromosome X [2 ]. Staining antibodies already established against the a5 chain of these a chains in the kidney and skin basement require denaturation treatment of cryostat sections to membranes (BM ) with monoclonal antibodies has expose their epitopes. To save time and eort for made it possible to diagnose Alport’s syndrome at the staining, a new epitope-defined monoclonal antibody protein level [5,6 ]. However, the staining procedure whose epitope is initially exposed on the surface of the with these antibodies involves acid‐urea treatment, molecule was established. incubation of sections with diluted animal serum to Methods. Two monoclonal antibodies against the avoid non-specific adsorption of secondary antibody triple-helical domains of the type IV collagen a2 and to the sections, incubation with primary antibody, and a5 chains were established with synthetic peptides as incubation with secondary antibody. The authors immunogens by the rat lymph node method. Their thought that this complicated, time-consuming and epitope were EAIQP at the positions of 675‐679 of labour-intensive staining was not a good method for the a2 chain, and IDVEF at the positions of 251‐255 the first step of examination at the time of a kidney of the a5 chain, respectively. They were purified with biopsy even when there is a suspicion of Alport’s synthetic peptide-coupled anity columns, and then syndrome. In place of the indirect staining, direct conjugated with Texas red and FITC, respectively. immunofluorescence staining with fluorochromeResults. The mixture of fluorochrome-conjugated anti- conjugated antibodies would be more suitable for this bodies was able to detect the distribution of the a2 purpose. and a5 chains in the normal and Alport kidney and The a5 (IV ) chain is the key chain in the diagnosis skin by direct immunofluorescence staining with and of Alport’s syndrome. First, in this syndrome, the X without denaturation treatment of the sections. chromosome-linked type is the major one, and is Conclusions. The direct double immunofluorescence induced by gene mutation of the a5( IV ) chain [7 ]. staining of kidney and skin cryostat sections with the Second, abnormal distribution of the a5 (IV ) chain can fluorochrome-conjugated antibodies is useful, reliable, be detected in the kidney of Alport’s patients at the and convenient for diagnosis of Alport’s syndrome. protein level in both X chromosome-linked Alport’s syndrome [5,6 ] and the autosomal recessive type [8 ] |
| Databáze: | OpenAIRE |
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