Table 2. All T cell hybridomas show equivalent baseline surface expression of CD8α and TCR, as well as IL-2 release in response to CD3 or PMA-Ca2+ ionophore stimulation.
| CDR3β sequence*
|
Surface expression†
|
IL-2 release‡
|
||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| TCR mutant | 95 | 96 | 97 | 98 | 99 | 100 | 101 | 102 | CD8α, % | TCR, % | PMA | CD3ε |
| WT | S | S | G | V | G | T | E | V | 91.1 | 93.6 | 55.2 | 99.3 |
| G97A | — | — | A | — | — | — | — | — | 96.9 | 85.1 | 46.9 | 83.7 |
| G97/99A | — | — | A | — | A | — | — | — | 85.3 | 94.9 | 47.3 | 72.4 |
| G99A | — | — | — | — | A | — | — | — | 95.9 | 100.0 | 56.4 | 83.3 |
| V98L | — | — | — | L | — | — | — | — | 100.0 | 87.7 | 53.9 | 85.3 |
| V98D | — | — | — | D | — | — | — | — | 93.4 | 92.7 | 47.9 | 83.5 |
*
CDR3β sequences of WT N30.7 TCR (Vβ13, Jβ1.1) and TCR mutants G97A, G97/99A, G99A, V98L, and V98D. The amino acid residue mutated is indicated.
†
CD8α and TCR surface expression was determined by FACS analysis with anti-CD8α and anti-TCRβ (percentage relative to the hybridoma with the highest surface expression).
‡
IL-2 release (units/ml) in response to either anti-CD3ε or PMA-Ca2+ ionophore stimulation.