Fig 1.

Activation of the human leucocyte antigen (HLA)-A2-restricted ILA1 CD8⁺ T cell clone with biophysically characterized altered peptide ligands (APLs). ILA1 cells were stimulated with peptide-pulsed HLA-A2⁺ C1R target cells as indicated. Five functional readouts [CD107a, macrophage inflammatory protein (MIP)-1β, tumour necrosis factor (TNF)-α, interferon (IFN)-γ and interleukin (IL)-2] were measured by flow cytometry. (a) Overview of functional profiles. Pie chart segments represent the fraction of cells expressing the number of functions indicated in the key. Serial gating was performed as follows: (i) doublets were excluded in a forward scatter (FSC)-A versus FSC-W display; (ii) artefacts and fluorochrome aggregates were removed by Boolean analysis; (iii) viable CD3⁺ CD8⁺ cells were selected; and (iv) individual functions were identified. Combination gates were exported into spice software version 5·33 for further analysis. (b) Detailed analysis of functional profiles at peptide concentrations of 10⁻⁵ M. The pie charts are extended with arcs defining expressed functions as indicated in the key. The index peptide is denoted in red. Data shown are representative of three independent experiments. The corresponding biophysical parameters are listed in Table 1.