Erratum

In1, there are errors in figures 1 to 5. In Figures 1 and 2, ‘TCR‐α GFP’ and ‘Concentration of peptide (μM)’ were incorrectly labelled as ‘TCR‐a GFP’ and ‘Concentration of peptide (mM)’, respectively, in the published version. In Figures 3, 4 and 5, the images published were poor in quality.

Figure 1.

Reconstruction of naturally isolated T cell receptor (TCR) pairs. (a) TCR‐deficient TG40 cells expressing the naturally isolated αβ TCR pairs A1‐T/B11‐J1 (TCR‐139), A1‐J31/B11 (H127) or A2‐R/B4 (S1) were stained with anti‐CD3 monoclonal antibody (mAb) (top row) or VY8/B35 tetramer (middle row) and analysed by flow cytometry. Mock‐transduced TG40 cells were used as negative controls. Tetramer staining is depicted as mean fluorescence intensity after gating on 7‐aminoactinomycin D (7‐AAD)^– green fluorescent protein (GFP)⁺CD31⁺ cells. To assess T cell sensitivity (bottom row), TCR genes were delivered into TG40 cells expressing human CD8α, as described previously [34]. CD3⁺ cells were then enriched using magnetic beads and analysed for interleukin (IL)‐2 secretion in response to various concentrations of VY8 peptide. The amount of IL‐2 obtained with mock‐transduced TG40 cells was consistently <5·0 U/ml and the amount of IL‐2 secreted by TCR‐transduced TG40 cells was expressed as % maximal, normalized with respect to the anti‐CD3 mAb‐mediated activation. Data are representative of duplicate assays. (b) TCR cross‐reactivity was evaluated against a library of peptides in which each position was substituted with every amino acid except cysteine to generate a total of 144 individual variants (5 µM per peptide). Antigen sensitivity was calculated as the percent of maximum response compared to anti‐CD3 mAb‐stimulated cells. Graphical representations showing relative preferences for amino acid residues at each position were generated using WebLogo 3 (http://weblogo.threeplusone.com/) [38]. Colours represent physicochemical properties: green, polar (G, S, T, Y and C); purple, neutral (Q and N); blue, basic (K, R and H); red, acidic (D and E); black, hydrophobic (A, V, L, I, P, W, F and M). The index residue at each position is outlined in yellow. Residue size is proportional to T cell recognition preference.

Figure 5.

Cross‐reactivity of hybrid T cell receptors (TCRs) in primary CD8⁺ T cells. Primary CD8⁺ T cells expressing hybrid TCRs were tested for preferred recognition residues at each position along the peptide backbone using an octamer combinatorial peptide library incorporating a total of 2·4 × 10¹⁰ different octamer peptides. In each of 160 peptide submixtures, one position was fixed with a defined amino acid residue and all other positions were degenerate, including any amino acid except cysteine. Macrophage inflammatory protein (MIP)‐1β release was quantified by enzyme‐linked immunosorbent assay (ELISA). Responses are shown as fold change MIP‐1β production relative to the lowest response at each position defined as 1·0. Background responses were 70 ± 8·9 pg/ml and the lowest response for CD8⁺ T cells expressing A1‐T, A1‐S and A1‐R at each position were 183 ± 26, 93 ± 44 and 122 ± 19 pg/ml, respectively. Fold changes >1·5 were considered positive. A representative set of duplicate assays is shown. Black bars depict residues corresponding to the VY8 index sequence.

Figure 2.

Reconstruction of hybrid T cell receptors (TCRs). (a–d) The indicated combinations of TCR‐α (A1‐T, A1‐S, A1‐R and A2‐R) and ‐β (B4, B11‐J1, B11‐J2 and B11) chains were analysed for cell surface expression of TCR/CD3, VY8/B35 tetramer binding and VY8 reactivity, as described in Fig. 1a.

Figure 3.

Cross‐reactivity of hybrid T cell receptors (TCRs). (a–c) The indicated combinations of TCR‐α (A1‐T, A1‐S and A1‐R) and ‐β (B11‐J1, B11‐J2 and B11) chains were analysed for cross‐reactivity, as described in Fig. 1b.

Figure 4.

Reconstruction of hybrid T cell receptors (TCRs) in primary CD8⁺ T cells. (a–c) Primary CD8⁺ T cells transduced with the indicated TCR pairs were examined for rCD2 expression (a), VY8/B35 tetramer staining after gating on 7‐aminoactinomycin D (7‐AAD)^–CD8⁺ cells (b) and macrophage inflammatory protein (MIP)‐1β production in response to peptide stimulation (c). In (a) and (b), data are representative of duplicate assays. In (b), the percentage and mean fluorescence intensity of live tetramer⁺ CD8⁺ T cells are indicated in each histogram. In (c), standard deviation from the mean of two replicates is shown.

The corrected figures are as follows.

We apologize for these errors.