IL-2 restores the ability of WAS and XLT NK cells to accumulate F-actin upon their interaction with sensitive targets: time course
. | Binding time . | . | . | . | . | . | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| . | Medium alone . | . | . | IL-2 . | . | . | |||||
| Sample . | 0 min . | 15 min . | 30 min . | 0 min . | 15 min . | 30 min . | |||||
| Healthy donors | |||||||||||
| Mean ± SD* | 52 ± 9 | 245 ± 33 (4.7 ± 0.4) | 257 ± 22 (5.1 ± 0.5) | 51 ± 4.7 | 252 ± 51 (5 ± 0.5) | 266 ± 28 (5.1 ± 0.7) | |||||
| Patients with XLT | |||||||||||
| W2 | 62 | 137 (2.2) | 143 (2.3) | 63 | 315 (5) | 283 (4.5) | |||||
| W3 | 45 | 94.5 (2.1) | 112 (2.5) | 59 | 242 (4.1) | 254 (4.3) | |||||
| W18 | 48 | 125 (2.6) | 139 (2.9) | 47 | 202 (4.3) | 174 (3.7) | |||||
| W19 | 46 | 106 (2.3) | 129 (2.8) | 57 | 234 (4.1) | 239 (4.2) | |||||
| Mean ± SD | 50 ± 7.9 | 115.6 ± 16 (2.3 ± 0.2) | 130.7 ± 12 (2.6 ± 0.3) | 56 ± 6.8 | 248 ± 41 (4.4 ± 0.4) | 237 ± 40 (4.2 ± 0.3) | |||||
| Patients with WAS | |||||||||||
| W4 | 45 | 81 (1.8) | 85 (1.9) | 55 | 236 (4.3) | 253 (4.6) | |||||
| W15 | 57 | 114 (2.0) | 125 (2.2) | 67 | ND | 275 (4.1) | |||||
| W22 | 65 | 104 (1.6) | 117 (1.8) | 70 | 224 (3.2) | 231 (3.3) | |||||
| Mean ± SD | 55 ± 10 | 99.6 ± 14 (1.8 ± 0.2) | 109 ± 17 (2.0 ± 0.2) | 64 ± 7.9 | 230 ± 6.0 (3.7 ± 0.8) | 253 ± 18 (4 ± 0.6) | |||||
. | Binding time . | . | . | . | . | . | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| . | Medium alone . | . | . | IL-2 . | . | . | |||||
| Sample . | 0 min . | 15 min . | 30 min . | 0 min . | 15 min . | 30 min . | |||||
| Healthy donors | |||||||||||
| Mean ± SD* | 52 ± 9 | 245 ± 33 (4.7 ± 0.4) | 257 ± 22 (5.1 ± 0.5) | 51 ± 4.7 | 252 ± 51 (5 ± 0.5) | 266 ± 28 (5.1 ± 0.7) | |||||
| Patients with XLT | |||||||||||
| W2 | 62 | 137 (2.2) | 143 (2.3) | 63 | 315 (5) | 283 (4.5) | |||||
| W3 | 45 | 94.5 (2.1) | 112 (2.5) | 59 | 242 (4.1) | 254 (4.3) | |||||
| W18 | 48 | 125 (2.6) | 139 (2.9) | 47 | 202 (4.3) | 174 (3.7) | |||||
| W19 | 46 | 106 (2.3) | 129 (2.8) | 57 | 234 (4.1) | 239 (4.2) | |||||
| Mean ± SD | 50 ± 7.9 | 115.6 ± 16 (2.3 ± 0.2) | 130.7 ± 12 (2.6 ± 0.3) | 56 ± 6.8 | 248 ± 41 (4.4 ± 0.4) | 237 ± 40 (4.2 ± 0.3) | |||||
| Patients with WAS | |||||||||||
| W4 | 45 | 81 (1.8) | 85 (1.9) | 55 | 236 (4.3) | 253 (4.6) | |||||
| W15 | 57 | 114 (2.0) | 125 (2.2) | 67 | ND | 275 (4.1) | |||||
| W22 | 65 | 104 (1.6) | 117 (1.8) | 70 | 224 (3.2) | 231 (3.3) | |||||
| Mean ± SD | 55 ± 10 | 99.6 ± 14 (1.8 ± 0.2) | 109 ± 17 (2.0 ± 0.2) | 64 ± 7.9 | 230 ± 6.0 (3.7 ± 0.8) | 253 ± 18 (4 ± 0.6) | |||||
NK cells from healthy donors and from patients with XLT or WAS, treated or not treated with human recombinant IL-2 (250 IU/mL) for 48 hours, were loaded with calceine am and then allowed to bind to K562 target cells (E/T ratio, 5:1) for the indicated times at 37°C. After stimulation, cells were fixed, permeabilized, and stained with rhodamine phalloidin, and the F-actin MFI was evaluated using FACS analysis in an equal number of binders. Results in parentheses indicate the F-actin MFI fold increase. Statistical analysis of the mean of fold increases of F-actin MFI of XLT and WAS NK cell binders, performed using the Student t test, indicates that the inhibition of F-actin accumulation observed in patients with XLT and WAS compared with those of control donors was statistically significant (P < .0001 at 15 and 30 minutes) and that IL-2 significantly restored this defect (P > .05).
ND indicates not determined.
Data are expressed as mean ± SD of F-actin MFI obtained in 6 healthy donors.