AID+ fraction and expression following in vitro stimulation
. | Cells/well . | . | . | . | . | . | . | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample . | 50 . | 100 . | 400 . | 800 . | 1600 . | 3200 . | QPCR value* . | |||||
| CLL203 | ||||||||||||
| Before | — | — | 1/93 | — | — | 0.28 | ||||||
| PMA + ionomycin | 3/95 | 7/95 | — | — | — | — | 4.78 | |||||
| αCD40 + IL-4 | 93/93 | 90/95 | — | — | — | — | 2398 | |||||
| CLL361 | ||||||||||||
| Before | — | — | — | 1/32 | 0/32 | 1/31 | 5.2 | |||||
| PMA + ionomycin | — | 0/32 | 1/32 | — | 1/30 | — | 11.8 | |||||
| αCD40 + IL-4 | — | 10/32 | 23/32 | — | 30/31 | — | 235 | |||||
| CLL127 | ||||||||||||
| Before | — | — | — | 0/32 | 1/32 | 0/31 | 0.5 | |||||
| PMA + ionomycin | — | 0/32 | 0/31 | — | 1/28 | — | 7.7 | |||||
| αCD40 + IL-4 | — | 2/32 | 2/32 | — | 5/31 | — | 116.8 | |||||
. | Cells/well . | . | . | . | . | . | . | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample . | 50 . | 100 . | 400 . | 800 . | 1600 . | 3200 . | QPCR value* . | |||||
| CLL203 | ||||||||||||
| Before | — | — | 1/93 | — | — | 0.28 | ||||||
| PMA + ionomycin | 3/95 | 7/95 | — | — | — | — | 4.78 | |||||
| αCD40 + IL-4 | 93/93 | 90/95 | — | — | — | — | 2398 | |||||
| CLL361 | ||||||||||||
| Before | — | — | — | 1/32 | 0/32 | 1/31 | 5.2 | |||||
| PMA + ionomycin | — | 0/32 | 1/32 | — | 1/30 | — | 11.8 | |||||
| αCD40 + IL-4 | — | 10/32 | 23/32 | — | 30/31 | — | 235 | |||||
| CLL127 | ||||||||||||
| Before | — | — | — | 0/32 | 1/32 | 0/31 | 0.5 | |||||
| PMA + ionomycin | — | 0/32 | 0/31 | — | 1/28 | — | 7.7 | |||||
| αCD40 + IL-4 | — | 2/32 | 2/32 | — | 5/31 | — | 116.8 | |||||
Summary of the results of the limiting dilution experiments performed on 3 CLL cases before and after 3.5 days of culture in the presence of either PMA and ionomycin or CD32-transfected L cells with anti-CD40 mAb and IL-4. The results are shown as a ratio of the AID+ wells to VH+ wells at indicated dilutions for each sample. In the last column, the real-time quantitative PCR values are reported. — indicates that no experiment was performed at those conditions.
Expression units, as per Figure 2.