American Society of Hematology

Figure 1

$Figure 1. NK-cell migration in response to chemokines is impaired in patients with XLT and patients with WAS. (A-B) Cultured NK cells from healthy donors or patients with WAS/XLT (W27, W28, W29, W19, W35) or highly purified freshly isolated W2 NK cells were assayed for their ability to migrate through ICAM-1 (5 μg/mL)–, or VCAM-1 (5 μg/mL)–precoated polycarbonate filters using CX3CL1/fractalkine (1nM) or CXCL12/SDF-1 (10nM) as chemoattractants. (C) Freshly isolated PBMCs from healthy donors or patients with WAS/XLT were allowed to migrate through a monolayer of TNFα (10 ng/mL)–activated endothelial cells on transwell filters in response to CXCL12/SDF-1 (1nM) or CX3CL1/fractalkine (1nM). After 1 hour of incubation, the number of migrated PBMCs was evaluated by FACS; the percentage of CD56+CD3− NK cells in input and migrated cells was assessed by immunofluorescence and FACS analysis. The percentage of migrated NK cells was calculated as follows: number of migrated NK cells/number of input NK cells × 100. Data from patients with WAS/XLT are expressed as the mean ± SD of the percentage of migrated cells obtained from 2 independent determinations. Control (Cntl) value represent the mean ± SD of the percentage of NK-cell migration obtained from 6 (A-B) or 4 healthy donors (C). ND indicates not determined. Statistical analysis performed by Student t test comparing the mean percentage of XLT/WAS NK-cell migration with that of control donors indicates that the inhibition of migration observed in all patients with WAS/XLT except for W35 is statistically significant (P < .02).$

NK-cell migration in response to chemokines is impaired in patients with XLT and patients with WAS. (A-B) Cultured NK cells from healthy donors or patients with WAS/XLT (W27, W28, W29, W19, W35) or highly purified freshly isolated W2 NK cells were assayed for their ability to migrate through ICAM-1 (5 μg/mL)–, or VCAM-1 (5 μg/mL)–precoated polycarbonate filters using CX3CL1/fractalkine (1nM) or CXCL12/SDF-1 (10nM) as chemoattractants. (C) Freshly isolated PBMCs from healthy donors or patients with WAS/XLT were allowed to migrate through a monolayer of TNFα (10 ng/mL)–activated endothelial cells on transwell filters in response to CXCL12/SDF-1 (1nM) or CX3CL1/fractalkine (1nM). After 1 hour of incubation, the number of migrated PBMCs was evaluated by FACS; the percentage of CD56⁺CD3⁻ NK cells in input and migrated cells was assessed by immunofluorescence and FACS analysis. The percentage of migrated NK cells was calculated as follows: number of migrated NK cells/number of input NK cells × 100. Data from patients with WAS/XLT are expressed as the mean ± SD of the percentage of migrated cells obtained from 2 independent determinations. Control (Cntl) value represent the mean ± SD of the percentage of NK-cell migration obtained from 6 (A-B) or 4 healthy donors (C). ND indicates not determined. Statistical analysis performed by Student t test comparing the mean percentage of XLT/WAS NK-cell migration with that of control donors indicates that the inhibition of migration observed in all patients with WAS/XLT except for W35 is statistically significant (P < .02).

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