Deep TCRB sequencing of tetramer-sorted cell populations from GS1-17A T-cell lines
| Cells* and frequency (%) . | Count . | TCRBV gene† . | TCRBD gene† . | TCRBJ gene† . | Junction sequence . | Sequence identifier‡ . |
|---|---|---|---|---|---|---|
| 17A-5YR-L1 Tet-neg (323 000) | ||||||
| 69.83 | 468 811 | 6-6§ | 2*02 | 2-5*01 | CASSDGLAGGWETQYF | Seq19 |
| 15.98 | 107 291 | 7-9|| | 2*01 | 1-1*01 | CASSPWKVTGNTEAFF | Seq20 |
| 11.89 | 79 822 | 12-3 or 12-4¶ | 2*01 | 2-7*01 | CASSFSQPGRRDEQYF | Seq21 |
| 0.64 | 4282 | 24-1*01 | 2*01 | 2-1*01 | CATSDLGTSGYNEQFF | Seq22 |
| 0.14 | 957 | 6-6§ | 1*01 | 1-2*01 | CASSYSDRGYGYTF | Seq23 |
| 0.13 | 846 | 7-2# | 1*01 | 2-7*01 | CASSLGAGGEQYF | Seq24 |
| 17A-5YR-L1 Tet-pos (307 000) | ||||||
| 50.99 | 134 032 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 34.53 | 90 759 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
| 11.81 | 31 040 | 6-6§ | 2*02 | 2-5*01 | CASSDGLAGGWETQYF | Seq19 |
| 0.64 | 1691 | 7-9|| | 2*01 | 1-1*01 | CASSPWKVTGNTEAFF | Seq20 |
| 0.50 | 1306 | 12-3 or 12-4¶ | 2*01 | 2-7*01 | CASSFSQPGRRDEQYF | Seq21 |
| 0.35 | 907 | 19†† | 1*01 | 1-4*01 | CASRDPHRVEKLFF | Seq27 |
| 0.10 | 269 | 24-1*01 | 2*01 | 2-1*01 | CATSDLGTSGYNEQFF | Seq22 |
| 17A-5YR-L2 Tet-pos-med‡‡(102 301) | ||||||
| 56.02 | 152 869 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 16.78 | 45 777 | 18*01 | Not identified | 2-1*01 | CASSPLPYSYNEQFF | Seq28 |
| 7.45 | 20 322 | 7-2# | 1*01 | 1-1*01 | CASSAGFPTEAFF | Seq29 |
| 7.10 | 19 366 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
| 4.16 | 11 351 | 7-2# | 1*01 | 2-7*01 | CASSLGAGGEQYF | Seq24 |
| 0.90 | 2444 | 5-1*01 or *02 | 1*01 | 1-2*01 | CASSLAGREGGYTF | Seq30 |
| 0.18 | 486 | 20-1** | 1*01 | 2-5*01 | CSARWTPGQGGETQYF | Seq31 |
| 0.11 | 296 | 3-1 or 3-2a | 1*01 | 1-1*01 | CASSQAGGNTEAFF | Seq32 |
| 17A-5YR-L3 Tet-pos (178 500) | ||||||
| 65.62 | 193 633 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 10.46 | 30 873 | 19†† | 1*01 | 1-2*01 | CASSTGTTSNYGYTF | Seq33 |
| 6.13 | 18 075 | 5-1*01 or *02 | 1*01 | 2-1*01 | CASSFQGVSYNEQFF | Seq34 |
| 4.86 | 14 327 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
| 4.79 | 14 127 | 7-2# | 1*01 | 2-7*01 | CASSLGAGGEQYF | Seq24 |
| 0.14 | 407 | 19†† | 1*01 | 1-4*01 | CASRDPHRVEKLFF | Seq27 |
| 17A-5YR-L4 Tet-pos (238 283) | ||||||
| 76.58 | 285 747 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 19.78 | 73 815 | 6-1*01 | 2*02 | 1-1*01 | CASSARADTEAFF | Seq35 |
| 1.45 | 5397 | 4-2*01 or *02 | 2*02 | 2-1*01 | CASSQEKREFDEQFF | Seq36 |
| 0.46 | 1729 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
| Cells* and frequency (%) . | Count . | TCRBV gene† . | TCRBD gene† . | TCRBJ gene† . | Junction sequence . | Sequence identifier‡ . |
|---|---|---|---|---|---|---|
| 17A-5YR-L1 Tet-neg (323 000) | ||||||
| 69.83 | 468 811 | 6-6§ | 2*02 | 2-5*01 | CASSDGLAGGWETQYF | Seq19 |
| 15.98 | 107 291 | 7-9|| | 2*01 | 1-1*01 | CASSPWKVTGNTEAFF | Seq20 |
| 11.89 | 79 822 | 12-3 or 12-4¶ | 2*01 | 2-7*01 | CASSFSQPGRRDEQYF | Seq21 |
| 0.64 | 4282 | 24-1*01 | 2*01 | 2-1*01 | CATSDLGTSGYNEQFF | Seq22 |
| 0.14 | 957 | 6-6§ | 1*01 | 1-2*01 | CASSYSDRGYGYTF | Seq23 |
| 0.13 | 846 | 7-2# | 1*01 | 2-7*01 | CASSLGAGGEQYF | Seq24 |
| 17A-5YR-L1 Tet-pos (307 000) | ||||||
| 50.99 | 134 032 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 34.53 | 90 759 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
| 11.81 | 31 040 | 6-6§ | 2*02 | 2-5*01 | CASSDGLAGGWETQYF | Seq19 |
| 0.64 | 1691 | 7-9|| | 2*01 | 1-1*01 | CASSPWKVTGNTEAFF | Seq20 |
| 0.50 | 1306 | 12-3 or 12-4¶ | 2*01 | 2-7*01 | CASSFSQPGRRDEQYF | Seq21 |
| 0.35 | 907 | 19†† | 1*01 | 1-4*01 | CASRDPHRVEKLFF | Seq27 |
| 0.10 | 269 | 24-1*01 | 2*01 | 2-1*01 | CATSDLGTSGYNEQFF | Seq22 |
| 17A-5YR-L2 Tet-pos-med‡‡(102 301) | ||||||
| 56.02 | 152 869 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 16.78 | 45 777 | 18*01 | Not identified | 2-1*01 | CASSPLPYSYNEQFF | Seq28 |
| 7.45 | 20 322 | 7-2# | 1*01 | 1-1*01 | CASSAGFPTEAFF | Seq29 |
| 7.10 | 19 366 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
| 4.16 | 11 351 | 7-2# | 1*01 | 2-7*01 | CASSLGAGGEQYF | Seq24 |
| 0.90 | 2444 | 5-1*01 or *02 | 1*01 | 1-2*01 | CASSLAGREGGYTF | Seq30 |
| 0.18 | 486 | 20-1** | 1*01 | 2-5*01 | CSARWTPGQGGETQYF | Seq31 |
| 0.11 | 296 | 3-1 or 3-2a | 1*01 | 1-1*01 | CASSQAGGNTEAFF | Seq32 |
| 17A-5YR-L3 Tet-pos (178 500) | ||||||
| 65.62 | 193 633 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 10.46 | 30 873 | 19†† | 1*01 | 1-2*01 | CASSTGTTSNYGYTF | Seq33 |
| 6.13 | 18 075 | 5-1*01 or *02 | 1*01 | 2-1*01 | CASSFQGVSYNEQFF | Seq34 |
| 4.86 | 14 327 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
| 4.79 | 14 127 | 7-2# | 1*01 | 2-7*01 | CASSLGAGGEQYF | Seq24 |
| 0.14 | 407 | 19†† | 1*01 | 1-4*01 | CASRDPHRVEKLFF | Seq27 |
| 17A-5YR-L4 Tet-pos (238 283) | ||||||
| 76.58 | 285 747 | 20-1** | 1*01 | 2-2*01 | CSARYFGPPKAGELFF | Seq25 |
| 19.78 | 73 815 | 6-1*01 | 2*02 | 1-1*01 | CASSARADTEAFF | Seq35 |
| 1.45 | 5397 | 4-2*01 or *02 | 2*02 | 2-1*01 | CASSQEKREFDEQFF | Seq36 |
| 0.46 | 1729 | 20-1** | 1*01 | 1-6*02 | CSARVQGGNSPLHF | Seq26 |
Cells were sorted with the DRB1*01:01-FVIII2194-2213 tetramer into populations that were stained (Tet-pos) or not stained (Tet-neg) with tetramer (supplemental Figure 5). The number of cells collected from each population is indicated in parentheses. Genomic DNA was isolated from cells, and deep TCRB-CDR3 sequencing was performed by Adaptive Biotechnologies. Sequences were analyzed with ImmunoSEQ Analyzer version 2.0. Productive sequences with a frequency ≥0.1% are shown.
Gene names for nucleotide sequences were determined using IMGT/V-QUEST version 3.3.5.50
Sequences were given a numerical identifier for labeling purposes. Analyzed nucleotide sequences are in supplemental Table 6.
Sequence matched equally well to 6-6*01, *02, *03, and *04 alleles.
Sequence matched equally well to 7-9*01, *02, *03, *04, and *05 alleles.
Sequence matched equally well to 12-3*01, 12-4*01, and 12-4*02 alleles.
Sequence matched equally well to 7-2*01, *02, *03, and *04 alleles.
Sequence matched equally well to 20-1*01, *02, *03, *04, and *05 alleles.
Sequence matched equally well to 19*01, *02, and *03 alleles.
Two tetramer-positive populations were observed at the time of sorting the 17A-5YR-L2 cell line. These populations were sorted and labeled medium and high, corresponding to their respective tetramer avidities. The high-avidity population (19 801 cells) was smaller than the medium-avidity population, and their sequence compositions were very similar. Therefore, only the medium-avidity population results are shown here.
Sequence matched equally well to 3-1*01 or *02, 3-2*01, *02, and *03 alleles.