Characteristics of B-CLL PBMCs used to test EBV-transformation conditions
| CLL ID . | Mutation status* . | IGHV mutation, %† . | IGHV . | IGHD . | IGHJ . | HCDR3 length‡ . | IgM+ wells, %§ . | Mean IgM level, μg/mL . |
|---|---|---|---|---|---|---|---|---|
| CLL246 | U | 0.3 | 1-69 | 3-3 | 6 | 21 | 100 | 3.3 |
| CLL487 | M | 4.8 | 2-05 | 1-1 | 4 | 17 | 68.8 | 0.36 |
| CLL493 | U | 0 | 1-69 | 5-5 | 4 | 21 | 96.3 | 0.24 |
| CLL526 | U | 0 | 1-69 | 3-3 | 6 | 23 | 100 | 2.12 |
| CLL698 | U | 0 | 1-69 | 3-3 | 6 | 21 | 100 | 5.81 |
| CLL816 | U | 0 | 1-69 | 3-3 | 6 | 25 | 95.8 | 0.17 |
| CLL821 | U | 0 | 1-69 | 3-3 | 4 | 22 | 100 | 0.29 |
| CLL822 | U | 0 | 2-05 | 6-13 | 6 | 25 | 100 | 0.30 |
| CLL939 | M | 6.6 | 3-66 | 3-10 | 3 | 11 | 50.4 | 0.05 |
| CLL1073 | U | 0 | 1-69 | 1-26 | 6 | 20 | 85.0 | 2.04 |
| CLL1075 | M | 5.9 | 1-69 | 5-12 | 4 | 16 | 100 | 1.63 |
| CLL1080 | M | 2.8 | 3-09 | 3-3 | 4 | 15 | 50.0 | 1.03 |
| CLL1223 | M | 6.2 | 2-05 | 3-9 | 4 | 17 | 85.0 | 2.03 |
| CLL1296 | M | 7.6 | 3-07 | 3-22 | 5 | 13 | 93.8 | 2.31 |
| CLL1324 | U | 0 | 1-69 | 3-10 | 6 | 23 | 100 | 2.53 |
| CLL ID . | Mutation status* . | IGHV mutation, %† . | IGHV . | IGHD . | IGHJ . | HCDR3 length‡ . | IgM+ wells, %§ . | Mean IgM level, μg/mL . |
|---|---|---|---|---|---|---|---|---|
| CLL246 | U | 0.3 | 1-69 | 3-3 | 6 | 21 | 100 | 3.3 |
| CLL487 | M | 4.8 | 2-05 | 1-1 | 4 | 17 | 68.8 | 0.36 |
| CLL493 | U | 0 | 1-69 | 5-5 | 4 | 21 | 96.3 | 0.24 |
| CLL526 | U | 0 | 1-69 | 3-3 | 6 | 23 | 100 | 2.12 |
| CLL698 | U | 0 | 1-69 | 3-3 | 6 | 21 | 100 | 5.81 |
| CLL816 | U | 0 | 1-69 | 3-3 | 6 | 25 | 95.8 | 0.17 |
| CLL821 | U | 0 | 1-69 | 3-3 | 4 | 22 | 100 | 0.29 |
| CLL822 | U | 0 | 2-05 | 6-13 | 6 | 25 | 100 | 0.30 |
| CLL939 | M | 6.6 | 3-66 | 3-10 | 3 | 11 | 50.4 | 0.05 |
| CLL1073 | U | 0 | 1-69 | 1-26 | 6 | 20 | 85.0 | 2.04 |
| CLL1075 | M | 5.9 | 1-69 | 5-12 | 4 | 16 | 100 | 1.63 |
| CLL1080 | M | 2.8 | 3-09 | 3-3 | 4 | 15 | 50.0 | 1.03 |
| CLL1223 | M | 6.2 | 2-05 | 3-9 | 4 | 17 | 85.0 | 2.03 |
| CLL1296 | M | 7.6 | 3-07 | 3-22 | 5 | 13 | 93.8 | 2.31 |
| CLL1324 | U | 0 | 1-69 | 3-10 | 6 | 23 | 100 | 2.53 |
U indicates unmutated type; and M, > 2% mutated compared with germline according to IMGT.27
IGHV mutation (%) was compared with germline according to IMGT.27
HCDR3 length indicates number of amino acid residues in the HCDR3.
To determine frequencies of IgM+ wells, PBMCs from B-CLL patients were plated at 5000 cells per well in the presence of irradiated J774A.1 cells (50 000 cells per well) and ODN2006 (12.5 μg/mL). The culture supernatants were analyzed for IgM production. For comparison, we used EBV-transformed B-cell cultures under the same condition from 20 healthy control PBMCs and found IgM production in 100% of the wells tested with an average IgM level of 6.7 μg/mL.