Open Access Research article

Decreased plasma levels of soluble CD18 link leukocyte infiltration with disease activity in spondyloarthritis

Tue W Kragstrup12, Babak Jalilian1, Malene Hvid13, Anders Kjærgaard4, René Østgård1, Berit Schiøttz-Christensen5, Anne G Jurik6, William H Robinson2, Thomas Vorup-Jensen1* and Bent Deleuran127

Author Affiliations

1 Department of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, DK-8000 Aarhus, Denmark

2 Division of Immunology and Rheumatology, Stanford University, 296 Campus Drive, Stanford, CA 94305, USA

3 Department of Clinical Medicine, Aarhus University, Brendstrupgårdsvej 100, DK-8200 Aarhus N Denmark

4 Department of Anaesthesiology, Aarhus University Hospital, Aarhus, Denmark

5 Aarhus Rheumatology Clinic, Skt. Clemens Torv 17, DK-8000 Denmark

6 Department of Radiology, Aarhus University Hospital, Aarhus, Denmark

7 Department of Rheumatology, Aarhus University Hospital, Aarhus, Denmark

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Arthritis Research & Therapy 2014, 16:R42  doi:10.1186/ar4471

Published: 4 February 2014

Additional files

Additional file 1: Table S1:

Associations at time of inclusion between plasma soluble CD18 (sCD18) levels in all patients with spondyloarthritis (SpA) and self-assessment scores after correction for age, disease duration, HLA-B27 status, treatment, and C-reactive protein (CRP).

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Additional file 2: Table S2:

Associations at time of inclusion between plasma soluble CD18 (sCD18) levels in all patients with spondyloarthritis (SpA) and clinical scores and test results after correction for age, disease duration, HLA-B27 status, treatment, and C-reactive protein (CRP).

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Additional file 3: Figure S1:

Confocal microscopy analysis of the ability of sCD11/CD18 complexes to bind intercellular adhesion molecule 1 (ICAM-1) expressed on the human umbilical vein cell line EA.hy926 or spondyloarthritis (SpA) fibroblast-like synoviocytes (FLSs). (A) Schematic of the cellular incubations. In step 1, adherent cells were incubated with 10 ng/mL tumor necrosis factor-alpha (TNFα), which increased the ICAM-1 expression. In step 2, a source of CD11/CD18 was added (that is, either NHS or supernatant from peripheral blood mononuclear cell (PBMC) culture). In step 3, biotinylated antibody recognizing ligand-binding activated CD11/CD18 (KIM127) was added followed by addition of fluorochrom-labelled streptavidin for detection with confocal microscopy. (B) Binding of soluble CD18 (sCD18) to TNFα-treated cells. As outlined above, in step 1, cells were treated with either TNFα or plain medium as a control. In step 2, either NHS or culture supernatant was used or plain medium was used as a control. In step 3, either the antibody to CD18 (KIM127) was used or biotinylated monoclonal IgG1 immunoglobulin was used as a control. Red staining indicated the binding of CD18, further indicated with white arrows. The positions of cell nuclei were located by 4′,6-diamidino-2-phenylindole (DAPI) staining, indicated in blue.

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Additional file 4: Figure S2:

Depletion of soluble CD18 (sCD18) by binding to intercellular adhesion molecule 1 (ICAM-1) expressed on the human umbelical vein cell line EA.hy926 or spondyloarthritis (SpA) fibroblast-like synoviocytes (FLSs). Culture medium supplemented with 50% (vol/vol) normal human serum (NHS) or 50% (vol/vol) synovial fluid mononuclear cell (SFMC) supernatant as sCD18 source were incubated with EA.hy926 or SPA FLS cells, each cell type either cultured in the supplemented media with 10 ng/mL tumor necrosis factor-alpha (TNFα) (to induce ICAM-1 expression) or in the supplemented media without further additions (“Medium”).

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