NGS analysis of the TCRVα21-Cα mRNA repertoire
| . | TCRVβ19+CD48+ . | TCRVβ19−CD48+ . | TCRVβ19+CD48− . | TCRVβ19−CD48− . | ||||
|---|---|---|---|---|---|---|---|---|
| Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | |
| Patients†‡ | ||||||||
| P3 | 43.3 | 0 (2 999) | 767.3 | 0 (5 495) | 17.6 | 0.21 (2763) | 711.4 | 0 (5627) |
| P4 | 306.8 | 0 (3 514) | 1300 | 0.16 (2 406) | 5.5 | 0 (4538) | ||
| P6 | 2.2 | 1.11 (499) | 598 | 0 (807) | 0 (921) | |||
| P22 | 93.4 | 0 (2 468) | 1100 | 0 (920) | 32.2 | 0 (2355) | ||
| P23 | 28.4 | 75.8 (2 970) | 359 | 0 (5 643) | 8.9 | 0 (4389) | 2.89 | 0 (1016) |
| P24 | 21.8 | 6.34 (4 457) | 414 | 0.56 (4 243) | 0.96 | 31.79 (585) | ||
| P25 | 96.9 | 1.87 (1 229) | 1150 | 0 (3 232) | 328 | 1.79 (782) | ||
| P26 | 21.6 | 1.05 (2 185) | 592 | 0 (874) | 18.6 | 0 (1176) | ||
| P27 | 46.7 | 0 (1 359) | 1100 | 0.06 (6 209) | 17 | 0.21 (910) | 306 | 0 (1973) |
| P28 | 86.6 | 0 (3 911) | 8.7 | 1.86 (5585) | ||||
| P29 | 54.3 | 0 (3 389) | 1370 | 0 (4 386) | 35.9 | 0 (3845) | ||
| Controls† | ||||||||
| N8 | 77.6 | 0.42 (10 669) | 840 | 0.18 (10 778) | ||||
| N9 | 160 | 0.16 (10 914) | 1250 | 0.14 (10 281) | ||||
| N10 | 150 | 0 (3 532) | 1800 | 0.09 (4 103) | ||||
| N11 | 128 | 0 (3 722) | 1200 | 0.06 (4 558) | ||||
| N12 | 90 | 0 (779) | 800 | 0 (715) | ||||
| N13 | 60 | 0 (6 976) | 1040 | 0 (11 929) | ||||
| N14 | 60 | 0 (5 704) | 1500 | 0.25 (79 369) | ||||
| N15 | 80 | 0.31 (2 248) | 1420 | 0.025 (3 945) | ||||
| . | TCRVβ19+CD48+ . | TCRVβ19−CD48+ . | TCRVβ19+CD48− . | TCRVβ19−CD48− . | ||||
|---|---|---|---|---|---|---|---|---|
| Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | Sorted cells, ×103 . | % TCRVα21Jα31 (no. of reads*) . | |
| Patients†‡ | ||||||||
| P3 | 43.3 | 0 (2 999) | 767.3 | 0 (5 495) | 17.6 | 0.21 (2763) | 711.4 | 0 (5627) |
| P4 | 306.8 | 0 (3 514) | 1300 | 0.16 (2 406) | 5.5 | 0 (4538) | ||
| P6 | 2.2 | 1.11 (499) | 598 | 0 (807) | 0 (921) | |||
| P22 | 93.4 | 0 (2 468) | 1100 | 0 (920) | 32.2 | 0 (2355) | ||
| P23 | 28.4 | 75.8 (2 970) | 359 | 0 (5 643) | 8.9 | 0 (4389) | 2.89 | 0 (1016) |
| P24 | 21.8 | 6.34 (4 457) | 414 | 0.56 (4 243) | 0.96 | 31.79 (585) | ||
| P25 | 96.9 | 1.87 (1 229) | 1150 | 0 (3 232) | 328 | 1.79 (782) | ||
| P26 | 21.6 | 1.05 (2 185) | 592 | 0 (874) | 18.6 | 0 (1176) | ||
| P27 | 46.7 | 0 (1 359) | 1100 | 0.06 (6 209) | 17 | 0.21 (910) | 306 | 0 (1973) |
| P28 | 86.6 | 0 (3 911) | 8.7 | 1.86 (5585) | ||||
| P29 | 54.3 | 0 (3 389) | 1370 | 0 (4 386) | 35.9 | 0 (3845) | ||
| Controls† | ||||||||
| N8 | 77.6 | 0.42 (10 669) | 840 | 0.18 (10 778) | ||||
| N9 | 160 | 0.16 (10 914) | 1250 | 0.14 (10 281) | ||||
| N10 | 150 | 0 (3 532) | 1800 | 0.09 (4 103) | ||||
| N11 | 128 | 0 (3 722) | 1200 | 0.06 (4 558) | ||||
| N12 | 90 | 0 (779) | 800 | 0 (715) | ||||
| N13 | 60 | 0 (6 976) | 1040 | 0 (11 929) | ||||
| N14 | 60 | 0 (5 704) | 1500 | 0.25 (79 369) | ||||
| N15 | 80 | 0.31 (2 248) | 1420 | 0.025 (3 945) | ||||
NGS analysis of the TCRVα21-Cα mRNA repertoire in flow-sorted TCRVβ19+ and TCRVβ19− T cells from PNH patients and normal controls (see also supplemental Figure 4).
mRNA, messenger RNA; NGS, next-generation sequencing.
Median age (range) patients vs controls: 36 years (28-59) vs 36 years (27-52).
Due to very low frequencies of GPI-negative T-cell fractions and small sample sizes, deep sequencing of TCRVα21-Cα amplicons was not possible for all 4 populations in all patients with PNH.
Represent productive TCR reads.