Serum samples of patients with rheumatoid arthritis contain a specific autoantibody to “denatured” aldolase A in the osteoblast-like cell line, MG-63

OBJECTIVE To identify rheumatoid arthritis (RA) specific autoantibody and its antigen in the human osteoblast-like cell line, MG-63. METHODS MG-63 cell extract was subjected to western blotting by using RA and normal serum samples as probes. The autoantigen was purified and its N-terminal sequence w...

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Bibliographic Details
Published in:Annals of the rheumatic diseases 1999-03, Vol.58 (3), p.169-174
Main Authors: Ukaji, Fumio, Kitajima, Isao, Kubo, Toshikazu, Shimizu, Choji, Nakajima, Toshihiro, Maruyama, Ikuro
Format: Article
Language:English
Subjects:
Adult
Aged
aldolase A
Antigens
Arthritis, Rheumatoid - enzymology
Arthritis, Rheumatoid - immunology
Autoantibodies - analysis
Autoantibodies - blood
autoantibody
Biological and medical sciences
Blotting, Western
Cell Line
denature
Diseases of the osteoarticular system
Enzyme-Linked Immunosorbent Assay
Enzymes
Extended Reports
Female
Fructose-Bisphosphate Aldolase - immunology
Humans
Immunoblotting
Immunoglobulins
Inflammatory joint diseases
Isoenzymes - immunology
Laboratories
Male
Medical sciences
Middle Aged
Osteoblasts - enzymology
Osteoblasts - immunology
Protein Denaturation
Proteins
Rheumatoid arthritis
Studies
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Summary:OBJECTIVE To identify rheumatoid arthritis (RA) specific autoantibody and its antigen in the human osteoblast-like cell line, MG-63. METHODS MG-63 cell extract was subjected to western blotting by using RA and normal serum samples as probes. The autoantigen was purified and its N-terminal sequence was determined by automated Edman degradation. The reactivity of denatured aldolase A was evaluated by immunoblotting. Screening by enzyme linked immunosorbent assay (ELISA) using the autoantibody was performed. RESULTS 40 kDa protein was found only in the RA serum samples and it was identified as aldolase A. A polyclonal antibody for rabbit muscle aldolase A bound to the 40 kDa protein and reacted in preference with the denatured enzyme. Using ELISA for denatured rabbit aldolase A, the autoantibody was found in approximately 10% of RA patients, whereas it was not found in the other arthropathy and healthy adults. CONCLUSION This 40 kDa anti-aldolase A autoantibody, which was identified only in serum samples of RA patients with severe bone erosion, could be related to a certain event that induces RA specific joint destructions.
ISSN:0003-4967
1468-2060
DOI:10.1136/ard.58.3.169