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. 1999 Feb;58(2):96–102. doi: 10.1136/ard.58.2.96

A quantitative method for detecting deposits of amyloid A protein in aspirated fat tissue of patients with arthritis

B Hazenberg 1, P Limburg 1, J Bijzet 1, M H van Rijswijk 1
PMCID: PMC1752828  PMID: 10343524

Abstract

OBJECTIVE—To describe a new, quantitative, and reproducible method for detecting deposits of amyloid A protein in aspirated fat tissue and to compare it with smears stained with Congo red.
METHODS—After extraction of at least 30 mg of abdominal fat tissue in guanidine, the amyloid A protein concentration was measured by a monoclonal antibody-based sandwich ELISA.
RESULTS—The concentrations in 24 patients with arthritis and AA amyloidosis (median 236, range 1.1-8530 ng/mg tissue) were higher (p<0.001) than in non-arthritic controls, uncomplicated rheumatoid arthritis, and other types of systemic amyloidosis (median 1.1, range 1.1-11.6 ng/mg tissue). Patients with extensive deposits, according to Congo red staining, had higher concentrations than patients with minute deposits.
CONCLUSION—This is a new, quantitative, and reproducible method for detecting deposits of amyloid A protein in aspirated fat tissue of patients with arthritis, even when minute deposits are present as detected in smears stained with Congo red.

 Keywords: amyloid A protein; fat tissue; arthritis

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Figure 1  .

Figure 1  

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The amyloid A protein concentration (ng/mg tissue) of aspirated fat tissue of 22 controls, 25 patients with rheumatoid arthritis without AA amyloidosis (RA), 24 patients with arthritis and AA amyloidosis (AA), and 25 patients with AL or ATTR amyloidosis (AL/ATTR). Horizontal lines indicate median values. The limit of detection is 1.1 ng/mg fat tissue. The asterisk (*) indicates the difference (p < 0.001) between the patients with arthritis and AA amyloidosis and each of the three other groups.

Figure 2  .

Figure 2  

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The amyloid A protein concentration of aspirated fat tissue (ng/mg tissue) of 24 patients with AA amyloidosis divided into four groups with increasing deposition of amyloid in fat smears. The deposition of amyloid was scored by staining the smears with Congo red. Horizontal lines indicate median values. The asterisk (*) indicates the difference (p < 0.05) between the eight patients with extensive deposits (3+) and the eight patients with minute deposits (1+). No difference was found between the six patients with moderate deposits (2+) and the other groups.

Figure 3  .

Figure 3  

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The serum amyloid A protein (SAA) levels (mg/l) of the 25 patients with rheumatoid arthritis without AA amyloidosis. Horizontal lines indicate median values. The asterisk (*) indicates the difference (p < 0.01) between the eight patients with a measurable amyloid A protein concentration of fat tissue (> 1.1 ng/mg) and the 17 patients without a measurable concentration (1.1 ng/mg).

Figure 4  .

Figure 4  

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SDS-PAGE of non-acute phase serum (lane 1), acute phase serum (lane 2), fat extracts (lane 3-8) of a healthy control (lane 3), of patients with arthritis and AA amyloidosis (lane 4, 6, and 8), of a patient with rheumatoid arthritis without AA amyloidosis (lane 5), and of a patient with AL amyloidosis (lane 7) run on 15% polyacrylamide gel, immunoblotted, detected with the monoclonal antibody Reu.86.5 raised against serum amyloid A, followed by a chemiluminescence technique. Rainbow molecular weight markers were chosen from 2.35 to 46 kDa (Amersham, Buckinghamshire, UK).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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