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. Author manuscript; available in PMC: 2010 Apr 20.
Published in final edited form as: Nat Med. 2009 Feb 22;15(3):313–318. doi: 10.1038/nm.1917

Figure 2.

Figure 2

Impaired function of αMβ2, α4β1 and αVβ3 integrins on subjects’ cells. (a) Adhesion of neutrophils isolated from subject 1 and a healthy control to fibrinogen and denatured ovalbumin in the presence or absence of 200 nM PMA. Specificity of adhesion was determined in the presence of an excess of αMβ2 ligand, neutrophil inhibitory factor (NIF). Adhesion of control resting neutrophils to ovalbumin was assigned a value of 100%. Data shown represent means ± s.d. (n = 3, ** P < 0.01). (b,c) Subject 1’s lymphocytes and neutrophils did not aggregate upon stimulation. Normal and subject lymphocytes (b) and neutrophils (c) were stimulated with 0.16 μM PMA and 1 μM fMLP, respectively. Representative photographs of lymphocytes were taken 25 min after stimulation (b). Scale bar, 100 μm. Neutrophil aggregation was measured 10 min after stimulation (c). (Means ± s.d.; n = 3, ** P < 0.01). (d) Neutrophil oxidative burst was impaired in subject 1. Superoxide release from neutrophils stimulated with PMA or opsonized zymosan (OZ) particles was measured as described in the Supplementary Methods (data represent means ± s.d.; n = 3, ** P < 0.01, * P < 0.05). (e) Subject 1’s lymphocytes, unlike control cells, did not adhere to intercellular adhesion molecule-1 in response to PMA (n = 3, ** P < 0.01). (f) Expression of the β2 integrin activation epitope on subject 1’s (left) and control (right) lymphocytes. Cells were stimulated with PMA (solid line) and analyzed for binding of mAb 24 by FACS analysis. The dotted line represents staining with an isotype-matched control mAb. (g) Binding of activation-dependent ligand WOW-1 Fab to lymphocytes from subject 2 and control cells was measured by FACS analysis in the presence or absence of PMA (200 nM). The data represent means ± s.d., n = 3, ** P < 0.01. (h) Peripheral blood mononuclear cells were isolated from the blood of a healthy control, the two subjects and their parents and immortalized. Adhesion of immortalized cells to fibrinogen in the presence or absence of 200 nM PMA, 1 mM EDTA and 200 nM RGD peptide is shown, as indicated. The data represent means ± s.d., n = 5, ** P < 0.01.