PIG-A mutations in cultured CD34+CD59+ and CD34+CD59− cells from 2 PNH patients and healthy volunteers
| Case no. . | No. of colonies4-150 . | Frequency of mutations4-151 . | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| CD34+CD59+cells . | CD34+CD59−cells . | CD34+CD59+cells . | CD34+CD59− cells . | |||||||||||||
| CFU-E BFU-E . | CFU-GM . | CFU-E BFU-E . | CFU-GM . | CFU-E BFU-E . | CFU-GM . | CFU-E BFU-E . | CFU-GM . | |||||||||
| Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | |
| PNH patients | ||||||||||||||||
| 2 | 28 | 45 | 32 | 50 | 102 | 105 | 76 | 91 | ||||||||
| Mutation 2-1‡ | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 18:20 | 14:20 | 17:20 | 12:20 |
| Mutation 2-2 | — | — | — | — | — | — | — | — | 1:20 | 1:20 | 0:20 | 0:20 | 2:20 | 3:20 | 0:20 | 0:20 |
| Mutation 2-3 | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 4:20 |
| 3 | 24 | 37 | 59 | 34 | 131 | 116 | 80 | 116 | ||||||||
| Mutation 3-14-153 | — | — | — | — | — | — | — | — | 1:20 | 0:20 | 0:20 | 1:20 | 18:20 | 15:20 | 15:20 | 12:20 |
| Mutation 3-2 | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 4:20 | 0:20 |
| Mutation 3-34-155 | — | — | — | — | — | — | — | — | 0:20 | 1:20 | 0:20 | 0:20 | 0:20 | 3:20 | 0:20 | 0:20 |
| Mutation 3-4 | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 4:20 | 0:20 |
| Healthy volunteers | ||||||||||||||||
| 1 | 92 | 98 | 95 | 120 | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 |
| 2 | 132 | 140 | 72 | 144 | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 |
| Case no. . | No. of colonies4-150 . | Frequency of mutations4-151 . | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| CD34+CD59+cells . | CD34+CD59−cells . | CD34+CD59+cells . | CD34+CD59− cells . | |||||||||||||
| CFU-E BFU-E . | CFU-GM . | CFU-E BFU-E . | CFU-GM . | CFU-E BFU-E . | CFU-GM . | CFU-E BFU-E . | CFU-GM . | |||||||||
| Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | Day 7 . | Day 14 . | |
| PNH patients | ||||||||||||||||
| 2 | 28 | 45 | 32 | 50 | 102 | 105 | 76 | 91 | ||||||||
| Mutation 2-1‡ | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 18:20 | 14:20 | 17:20 | 12:20 |
| Mutation 2-2 | — | — | — | — | — | — | — | — | 1:20 | 1:20 | 0:20 | 0:20 | 2:20 | 3:20 | 0:20 | 0:20 |
| Mutation 2-3 | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 4:20 |
| 3 | 24 | 37 | 59 | 34 | 131 | 116 | 80 | 116 | ||||||||
| Mutation 3-14-153 | — | — | — | — | — | — | — | — | 1:20 | 0:20 | 0:20 | 1:20 | 18:20 | 15:20 | 15:20 | 12:20 |
| Mutation 3-2 | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 4:20 | 0:20 |
| Mutation 3-34-155 | — | — | — | — | — | — | — | — | 0:20 | 1:20 | 0:20 | 0:20 | 0:20 | 3:20 | 0:20 | 0:20 |
| Mutation 3-4 | — | — | — | — | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 4:20 | 0:20 |
| Healthy volunteers | ||||||||||||||||
| 1 | 92 | 98 | 95 | 120 | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 |
| 2 | 132 | 140 | 72 | 144 | — | — | — | — | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 | 0:20 |
CFU-E, BFU-E, and CFU-GM colonies were cultured for 7 or 14 days as described in “Patients, materials, and methods.” The numbers of colonies were counted using an inverted microscope in 2 or 3 dishes for each cultured cell population. The mean values are presented.
The numbers indicate the ratio of mutated clones to the total examined in each cultured cell population.
Mutation 2-1 in CFU-E, BFU-E, 7-day CFU-GM, and 14-day CFU-GM colonies from CD34+CD59− cells was predominant compared with mutations 2-2 and 2-3 (P < .0001 versus mutations 2-2 and 2-3; P < .001 versus mutation 2-2 and P < .0001 versus mutation 2-3; P< .0001 versus mutations 2-2 and 2-3; P < .0001 versus mutation 2-2 and P < .05 versus mutation 2-3, respectively).
Mutation 3-1 in CFU-E, BFU-E, 7-day CFU-GM, and 14-day CFU-GM colonies from CD34+CD59− cells was predominant compared with the other mutations (P < .0001 versus mutations 3-2, 3-3 and 3-4; P < .0001 versus mutations 3-2 and 3-4 and P < .001 versus mutation 3-3;P < .002 versus mutations 3-2 and 3-4 andP < .0001 versus mutation 3-3; P < .0001 versus mutations 3-2, 3-3 and 3-4, respectively).
Mutation 3-3 in BFU-E colonies from CD34+CD59− cells was predominant compared with mutations 3-2 and 3-4 (P < .05).