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. 2002 Dec;61(12):1047–1054. doi: 10.1136/ard.61.12.1047

Receptor activator NF-κB ligand (RANKL) expression in synovial tissue from patients with rheumatoid arthritis, spondyloarthropathy, osteoarthritis, and from normal patients: semiquantitative and quantitative analysis

T Crotti 1, M Smith 1, H Weedon 1, M Ahern 1, D Findlay 1, M Kraan 1, P Tak 1, D Haynes 1
PMCID: PMC1753975  PMID: 12429533

Abstract

Objectives: To compare receptor activator of NF-κB ligand (RANKL) production in the synovial tissue from patients with active rheumatoid arthritis (RA), inactive RA, spondyloarthropathies (SpA), osteoarthritis, and from normal subjects. In addition, to establish the cell lineages expressing RANKL in these tissues.

Methods: Immunohistological analysis of frozen synovial tissue biopsy specimens was performed using a monoclonal antibody (mAb) to detect RANKL. Sections were evaluated by computer assisted image analysis and semiquantitative analysis to compare RANKL expression between groups. Dual and sequential labelling with mAb RANKL and cell lineage specific monoclonal antibodies were used to determine the types of cells expressing RANKL.

Results: Higher levels of RANKL were expressed in tissues from patients with active RA and SpA than in tissues from patients with inactive RA, osteoarthritis, and from normal subjects. RANKL protein was associated with CD3 antigen-positive lymphocytes and some macrophages. RANKL was predominantly associated with activated, memory T cells (CD45Ro positive cells) in patients with active RA and spondyloarthropathy (SpA).

Conclusions: The highest levels of RANKL were detected in patients with RA with active synovitis and in some patients with SpA. An increase in RANKL in the inflamed joint of patients with RA, produced by infiltrating activated T cells and macrophages, is likely to be an important cause of joint erosions in RA.

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Figure 1 .

Figure 1

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RANKL detected with mAb 626/immunoperoxidase and AEC (red), in synovial tissue from a patient with RA with (A) active disease and (B) inactive disease, (C) a patient with SpA, and (D) a patient with OA. (E) to (H) identify cell lineages expressing RANKL (mAb 626/immunoperoxidase (red)) using dual immunohistochemical labelling on serial synovial biopsy sections from a patient with RA with active disease. RANKL expression is identified by mAb 626 using an AEC/immunoperoxidase technique combined with an immunohistochemical alkaline phosphatase technique using fast blue to identify cell surface markers. (E) Dual labelling with anti-CD3 (blue) and both (purple), (F) B cell antibody anti-CD22 (blue), (G) monocyte/macrophage lineage anti-CD68 (blue) and both (purple), (H) FLS with anti- CD55/fast blue (blue). All magnifications taken at x20.

Figure 2 .

Figure 2

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Panels (A), (C), (E), (G) demonstrate RANKL expression, detected with mAb 626/ immunoperoxidase and AEC (red), in synovial tissue from a patient with RA with active disease (A), inactive disease (C), SpA (E), and from a normal subject (G). Panels (B), (D), (F), (H) show sequential sections where CD45Ro is detected with immunoperoxidase and AEC (red) in synovial tissue from patients with active RA (B), inactive RA (D), SpA (F), and from a normal subject (H). All magnifications taken at x20.

Selected References

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