Chronic TCR-MHC (self)-interactions limit the functional potential of TCR affinity-increased CD8 T lymphocytes

摘要

Baseline expression levels of the TCR/CD3 complex, CD5 and c-CBL and phosphorylation levels of CD3ζ and ERK upon stimulation in relation to HLA-A2. Expression levels of TCR/CD3 complex and CD28 in A2 and A2 primary CD8 T cells ( ) or A2 and A2 J76 CD8αβ cells ( ) engineered with TCRs of incremental affinities and analyzed under steady-state culture condition, in the absence of cognate antigen. Quantification of CD3ζ and ERK1/2 phosphorylation in TCR-expressing A2 and A2 CD8αβ J76 cells after stimulation with NY-ESO-1 multimer, OKT3 (anti-CD3ε) antibody or PMA/Ionomycin. Quantification of CD5 and c-CBL expression in A2 and A2 CD8αβ J76 cells. Direct comparison (bottom panel) of c-CBL expression in A2 versus A2 CD8αβ J76 cells for the indicated TCR variants by two-tailed, paired test. Data are means ± SD and representative of 5 to 15 independent experiments. TCR-transduced A2 cells versus A2 T cells are depicted as red versus blue symbols. *  ≤ 0.05, **  ≤ 0.01, ***  ≤ 0.001 and ****  ≤ 0.0001