Varying the MHC-I affinity, TCR affinity or antigen dose alters the phenotype of CD8 T cells ex vivo. Varying the MHC-I affinity, TCR affinity or antigen.

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Varying the MHC-I affinity, TCR affinity or antigen dose alters the phenotype of CD8 T cells ex vivo. Varying the MHC-I affinity, TCR affinity or antigen dose alters the phenotype of CD8 T cells ex vivo. Mouse splenocytes (OT-1) were collected and immediately stimulated with SIINFEKL (OVA) or one of the variants that alters the MHC-I affinity (top), TCR affinity (middle) or different concentrations of SIINFEKL (bottom). The CD3 (TCR) levels, activation, and checkpoint receptor expression of CD8 T cells were monitored every 24 hours for 96 hours. Results are representative of two independent experiments with N = 3 mice each. NS, nonspecific peptide, SSX2 p103-111 (RLQGISPKI). Statistical analyses compare OVA with FT (top), OVA to NG (middle), and 2 μg/mL to 0.1 ng/mL OVA (bottom). For CD3 levels, OVA, FT, NG, and 0.1 ng/mL OVA were compared with NS. *, P < 0.05, two-way ANOVA followed by Bonferroni correction. Representative flow cytometry gating strategy is shown in Supplementary Fig. S3. Christopher D. Zahm et al. Cancer Immunol Res 2017;5:630-641 ©2017 by American Association for Cancer Research