Detection of AA76, a Common Form of Amyloid A Protein, as a Way of Diagnosing AA Amyloidosis.
Reactive amyloid deposits consist of amyloid A (AA) proteins, the degradation products of serum amyloid A (SAA). Since the most common species of AA is the amino terminal portion produced by cleavage between residues 76 and 77 of SAA (AA76), the presence of AA76 in tissues could be a consequence of AA amyloid deposition. This study assessed the diagnostic significance of the detection of AA76 for AA amyloidosis using two different approaches. Biopsy specimens (n=130 from 54 subjects) from gastroduodenal mucosa or abdominal fat (n=9 from 9 subjects) of patients who had already been diagnosed with or were suspected of having AA amyloidosis were used. Fixed mucosal sections were subjected to immunohistochemistry using a newly developed antibody recognizing the carboxyl terminal end of AA76 (anti-AA76). The non-fixed materials from gastroduodenal mucosa or abdominal fat were subjected to immunoblotting for detection of the size of AA76. Among the gastroduodenal specimens (n=115) from already diagnosed patients, the positive rates of Congo red staining, immunohistochemistry using anti-AA76, and immunoblotting were 68.4%, 73.0%, and 92.2%, respectively. The anti-AA76 did not stain the supposed SAA in the blood or leakage, which was stained by anti-SAA antibody. AA76 was not detected either by immunohistochemistry or by immunoblot in the materials from patients in whom AA amyloidosis had been ruled out. In the abdominal fat, the immunoblot detected AA76 in 8 materials from 8 already diagnosed patients and did not in 1 patient whose gastroduodenal mucosa was negative. In conclusion, the detection of AA76 may alter the ability to diagnose AA amyloidosis. In immunohistochemistry for fixed specimens, the new anti-AA76 antibody can improve the specificity. Immunoblot for non-fixed materials, which can considerably improve the sensitivity, should be beneficial for small materials like abdominal fat.
Department of Clinical Laboratory Medicine, Jichi Medical University, Tochigi, Japan Biochemical Research Laboratory, Eiken Chemical Co., Ltd., Tochigi, Japan.
Junji
J
Sato
Centre for Rheumatology, Dohgo Spa Hospital, Ehime, Japan.
Yasuaki
Y
Okuda
Health Service Center, Niigata University, Niigata, Japan.
Takeshi
T
Kuroda
Department of Clinical Laboratory Medicine, Jichi Medical University, Tochigi, Japan toshiyam@jichi.ac.jp.
Toshiyuki
T
Yamada
D000368:Aged; D000369:Aged, 80 and over; D000686:Amyloidosis / Q000175:diagnosis*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology; D004386:Duodenum / Q000473:pathology; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000473:pathology; D008875:Middle Aged; D000685:Serum Amyloid A Protein / Q000378:metabolism*; D013270:Stomach / Q000473:pathology
D000911:Antibodies, Monoclonal; D000685:Serum Amyloid A Protein
AA amyloidosis; abdominal fat; immunoblot; monoclonal antibody; serum amyloid A