Clinical features of CLL samples used in this study and ERK1/2, intracellular Ca2+, and BIM phosphorylation responses
| Sample . | IGHV status . | Binet stage at diagnosis . | Treatment required* . | CD38† . | ZAP-70† . | Maximum iCa2+ response‡ . | Maximum P-ERK1/2 response§ . | Maximum P-BIM response‖ . | |
|---|---|---|---|---|---|---|---|---|---|
| BIML . | BIMEL . | ||||||||
| 189 | M | A | Y | − | − | 17 | 1.5 | ND | ND |
| 233 | U | A | N | + | + | 71 | 1.2 | 23 | 39 |
| 238 | U | A | Y | − | + | 2 | 1.4 | 61 | 62 |
| 241 | M | A | N | − | − | 2 | 1.0 | 2 | 2 |
| 251 | M | A | N | − | − | 0 | 1.0 | 9 | 11 |
| 256 | U | B | Y | − | − | 20 | 1.8 | 59 | 75 |
| 261 | M | A | N | − | − | 0 | 1.0 | 0 | 5 |
| 266 | U | C | Y | + | + | 4 | ND | ND | ND |
| 268 | M | A | N | − | − | 30 | 1.4 | 0 | 17 |
| 271 | M | A | N | − | + | 0 | ND | 0 | 0 |
| 277 | M | NA | Y | − | − | 6 | 2.1 | 18 | 58 |
| 285 | U | A | Y | + | + | 39 | 1.5 | ND | ND |
| 291 | U | NA | NA | − | + | 16 | 1.5 | 29 | 48 |
| 292 | M | A | N | − | − | 0 | ND | 0 | 0 |
| 293 | U | A | Y | − | + | 0 | ND | 31 | 64 |
| 298 | U | C | NA | + | + | 14 | 0.7 | 44 | 43 |
| 299 | M | A | N | − | − | 0 | 1.2 | 0 | 68 |
| 303 | M | NA | NA | − | NA | 9 | ND | ND | ND |
| 304 | U | B | N | + | + | 33 | 2.4 | 31 | 70 |
| 309 | M | NA | NA | − | − | 55 | ND | ND | ND |
| 313 | M | A | Y | − | − | 9 | ND | ND | ND |
| 321 | U | NA | NA | − | NA | 22 | ND | ND | ND |
| 328 | U | A | N | − | − | 74 | 1.6 | 48 | 57 |
| 332 | M | NA | NA | − | − | 2 | ND | 20 | 53 |
| 333 | M | A | Y | − | − | 77 | 1.6 | 41 | 71 |
| 343 | U | A | Y | + | − | 77 | 1.7 | 58 | 79 |
| 351 | M | A | NA | + | − | 2 | ND | 1 | 2 |
| 357 | M | A | N | + | − | 2 | ND | 3 | 14 |
| 362 | M | A | N | − | − | 2 | ND | 8 | 21 |
| 363 | M | A | N | + | − | 5 | 1.4 | 5 | 59 |
| 370 | M | NA | NA | − | − | 3 | 1.8 | 11 | 25 |
| 374 | M | A | N | − | − | 55 | 1.5 | 0 | 0 |
| 376 | U | A | Y | + | − | 23 | 1.8 | 51 | 81 |
| 379 | M | A | Y | − | − | 11 | 1.4 | 41 | 67 |
| 383 | U | A | Y | − | − | 14 | 2.0 | 36 | 76 |
| 392 | M | A | N | − | − | 72 | ND | ND | ND |
| 394 | U | A | N | + | + | 46 | ND | ND | ND |
| 399 | M | A | N | − | − | 0 | ND | ND | ND |
| 410 | U | NA | NA | + | − | 25 | ND | ND | ND |
| 431 | M | NA | NA | − | − | ND | ND | ND | ND |
| Sample . | IGHV status . | Binet stage at diagnosis . | Treatment required* . | CD38† . | ZAP-70† . | Maximum iCa2+ response‡ . | Maximum P-ERK1/2 response§ . | Maximum P-BIM response‖ . | |
|---|---|---|---|---|---|---|---|---|---|
| BIML . | BIMEL . | ||||||||
| 189 | M | A | Y | − | − | 17 | 1.5 | ND | ND |
| 233 | U | A | N | + | + | 71 | 1.2 | 23 | 39 |
| 238 | U | A | Y | − | + | 2 | 1.4 | 61 | 62 |
| 241 | M | A | N | − | − | 2 | 1.0 | 2 | 2 |
| 251 | M | A | N | − | − | 0 | 1.0 | 9 | 11 |
| 256 | U | B | Y | − | − | 20 | 1.8 | 59 | 75 |
| 261 | M | A | N | − | − | 0 | 1.0 | 0 | 5 |
| 266 | U | C | Y | + | + | 4 | ND | ND | ND |
| 268 | M | A | N | − | − | 30 | 1.4 | 0 | 17 |
| 271 | M | A | N | − | + | 0 | ND | 0 | 0 |
| 277 | M | NA | Y | − | − | 6 | 2.1 | 18 | 58 |
| 285 | U | A | Y | + | + | 39 | 1.5 | ND | ND |
| 291 | U | NA | NA | − | + | 16 | 1.5 | 29 | 48 |
| 292 | M | A | N | − | − | 0 | ND | 0 | 0 |
| 293 | U | A | Y | − | + | 0 | ND | 31 | 64 |
| 298 | U | C | NA | + | + | 14 | 0.7 | 44 | 43 |
| 299 | M | A | N | − | − | 0 | 1.2 | 0 | 68 |
| 303 | M | NA | NA | − | NA | 9 | ND | ND | ND |
| 304 | U | B | N | + | + | 33 | 2.4 | 31 | 70 |
| 309 | M | NA | NA | − | − | 55 | ND | ND | ND |
| 313 | M | A | Y | − | − | 9 | ND | ND | ND |
| 321 | U | NA | NA | − | NA | 22 | ND | ND | ND |
| 328 | U | A | N | − | − | 74 | 1.6 | 48 | 57 |
| 332 | M | NA | NA | − | − | 2 | ND | 20 | 53 |
| 333 | M | A | Y | − | − | 77 | 1.6 | 41 | 71 |
| 343 | U | A | Y | + | − | 77 | 1.7 | 58 | 79 |
| 351 | M | A | NA | + | − | 2 | ND | 1 | 2 |
| 357 | M | A | N | + | − | 2 | ND | 3 | 14 |
| 362 | M | A | N | − | − | 2 | ND | 8 | 21 |
| 363 | M | A | N | + | − | 5 | 1.4 | 5 | 59 |
| 370 | M | NA | NA | − | − | 3 | 1.8 | 11 | 25 |
| 374 | M | A | N | − | − | 55 | 1.5 | 0 | 0 |
| 376 | U | A | Y | + | − | 23 | 1.8 | 51 | 81 |
| 379 | M | A | Y | − | − | 11 | 1.4 | 41 | 67 |
| 383 | U | A | Y | − | − | 14 | 2.0 | 36 | 76 |
| 392 | M | A | N | − | − | 72 | ND | ND | ND |
| 394 | U | A | N | + | + | 46 | ND | ND | ND |
| 399 | M | A | N | − | − | 0 | ND | ND | ND |
| 410 | U | NA | NA | + | − | 25 | ND | ND | ND |
| 431 | M | NA | NA | − | − | ND | ND | ND | ND |
Because of limitations in the amount of material, we were unable to perform all assays on all samples.
M indicates mutated; U, unmutated; NA, not available; Y, yes; N, no; and ND, not done.
Requirement for treatment at any point after diagnosis.
Samples with > 30% CD38+ or > 20% ZAP-70+ CD19+ cells were considered positive.
Maximum anti-IgM–induced intracellular Ca2+ response, as described.10 Values represent percentage of cells fluxing calcium over unstimulated levels. The cut-off for classification as positive signaler was 5%.
Maximum anti-IgM–induced ERK1/2 phosphorylation (fold induction compared with untreated cells; flow cytometry).
Maximum percentage of anti-IgM–induced BIM phosphorylation (immunoblotting).