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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Kragstrup et al. Arthritis Research & Therapy 2014 16:R42 doi:10.1186/ar4471