DRB1*01:01-FVIII2194-2213 tetramer staining avidity and TCRB gene sequence analysis for T-cell clones isolated from severe HA subject GS1-56A
| Clone* . | Tetramer staining† . | TCRBV gene‡ . | TCRBD gene‡ . | TCRBJ gene‡ . | V-D-J junction‡,§ . | Sequence identifier|| . |
|---|---|---|---|---|---|---|
| 56A-C1 | 974 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C2 | 643 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C3 | 7 | 18*01 | 2*01 or 2*02¶ | 2-7*01 | CASSAGLAETYEQYF | Seq02 |
| 56A-C4 | 869 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C6 | 896 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C7 | 613 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C8# | 202 | 7-8*01 or *03 | 2*01 or 2*02 | 2-3*01 | CASSSAPRLTSGRTDTQYF | Seq03 |
| 56A-C9 | 708 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C10 | 3 | 3-1 or 3-2 | 1*01 | 1-5*01 | CASSQGHGNQPQHF | Seq04 |
| 56A-C11 | 909 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C12 | 907 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C13 | 26 | 20-1** | 1*01 | 2-7*01 | CSARGQGAYEQYF | Seq05 |
| 56A-C14 | 852 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C15 | 5 | 9†† | 1*01 | 1-2*01 | CASSVAGTGPQVLYGYTF | Seq06 |
| 56A-C16 | 648 | nd | nd | nd | nd | |
| 56A-C17 | 804 | nd | nd | nd | nd | |
| 56A-C18 | 402 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| Clone* . | Tetramer staining† . | TCRBV gene‡ . | TCRBD gene‡ . | TCRBJ gene‡ . | V-D-J junction‡,§ . | Sequence identifier|| . |
|---|---|---|---|---|---|---|
| 56A-C1 | 974 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C2 | 643 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C3 | 7 | 18*01 | 2*01 or 2*02¶ | 2-7*01 | CASSAGLAETYEQYF | Seq02 |
| 56A-C4 | 869 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C6 | 896 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C7 | 613 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C8# | 202 | 7-8*01 or *03 | 2*01 or 2*02 | 2-3*01 | CASSSAPRLTSGRTDTQYF | Seq03 |
| 56A-C9 | 708 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C10 | 3 | 3-1 or 3-2 | 1*01 | 1-5*01 | CASSQGHGNQPQHF | Seq04 |
| 56A-C11 | 909 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C12 | 907 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C13 | 26 | 20-1** | 1*01 | 2-7*01 | CSARGQGAYEQYF | Seq05 |
| 56A-C14 | 852 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
| 56A-C15 | 5 | 9†† | 1*01 | 1-2*01 | CASSVAGTGPQVLYGYTF | Seq06 |
| 56A-C16 | 648 | nd | nd | nd | nd | |
| 56A-C17 | 804 | nd | nd | nd | nd | |
| 56A-C18 | 402 | 27*01 | Not identified | 1-1*01 | CASSFDPLNTEAFF | Seq01 |
nd, not determined.
56A-C5 is not included in the list because these cells did not expand sufficiently to permit tetramer staining.
Tetramer staining was quantitated from the MFI of staining with DRB1*01:01-FVIII2194-2213 tetramer minus MFI of staining with an irrelevant tetramer (DRB1*01:01-FVIII2178-2197). Results are from a single determination that occurred while screening clones that were expanding after single cell sorting.
Nucleotide sequences were analyzed using Adaptive Biotechnologies ImmunoSEQ Analyzer version 2.0. Gene names were determined using international ImMunoGeneTics information system/V-QUEry and STandardization (IMGT/V-QUEST) version 3.3.4.50 TCR nomenclature is in accordance with the international reference IMGT/GENE-DB.51
V-D-J junction sequence corresponds to the third hypervariable region of the TCR beginning with the second conserved cysteine encoded by the 3′ portion of the V-region and ends with the conserved phenylalanine encoded by the 5′ portion of the J-region.
Sequences were given a numerical identifier for labeling purposes. Analyzed nucleotide sequences are in supplemental Table 6.
Multiple gene names result from >1 known annotated V, D, or J gene sequence matching equally well with the aligned sequence.
56A-C8 expressed a second TCRB gene that corresponded to TCRBV27*01, TCRBD2*01 or *02, TCRBJ2-1*01 and is out of frame.
Sequence is identical to 20-1*01, *02, *03, *04, and *05 alleles.
Sequence is identical to 9*01, *02, and *03 alleles.