Figure 3.
Most γδ T-cell clones that respond to EBV express a Vγ9JγP CDR3. Two hundred thousand γδ T cells were isolated from PBMCs before and after stimulation with Akata, an EBV-transformed B cell line. (A) Pie charts show the γ-chain CDR3 repertoires of unstimulated γδ T cells and EBV-stimulated γδ T cells from 1 representative group 1 donor and 1 representative group 2 donor. CDR3γ V9-JP sequences are represented in blue and CDR3γ V9-J2 sequences are represented in red. The gray zone represents the combination of all other unique CDR3s within the CDR3γ repertoires. (B) Pie charts show the distribution of the 20 most abundant γ-chain CDR3s of unstimulated γδ T cells and EBV-stimulated γδ T cells, from 3 representative group 1 donors and 3 representative group 2 donors. The public CDR3s ALWEVQELGKKIKV (dark blue) and ALWEVRELGKKIKV (light blue) are shown. For each donor, the 5 most dominant CDR3s expressed by unstimulated γδ T cells are shown in different shades of color and, when required, in dark blue or light blue for ALWEVQELGKKIKV and ALWEVRELGKKIKV, respectively. To track these same CDR3s in the EBV-stimulated γδ T-cell repertoires, we depicted them in similar colors. The other 15 CDR3s are colored in light green and the remaining unique CDR3s are shaded in gray. (C) Kullback-Leibler sequence logos show the residue preferences for the dominant 14-mer CDR3γ sequences from EBV-stimulated γδ T cells of group 1 (N = 10 CDR3γ) and group 2 (N = 10 CDR3γ) donors. Populations of γδ T cells were isolated from EBV-stimulated PBMCs. Each logo consists of amino acid stacks, 1 for each position. The stack height indicates the sequence conservation; the residue height indicates its relative frequency. Residue colors correspond to biochemical properties: acidic (red), basic (blue), hydrophobic (black), polar (green). Logos were generated using WebLogo.28 (D) The charts illustrate the distribution of the 20 most abundant γ-chain CDR3s among unstimulated γδ T cells and EBV-stimulated γδ T cells from a group 1 donor: donor 1. Populations of γδ T cells were isolated from unstimulated PBMCs and EBV-stimulated PBMCs. CDR3γ V9-JP sequences are represented by different shades of blue and CDR3γ V9-J2 are represented by different shades of red. The gray zone indicates a combination of all the other unique CDR3s within the CDR3γ repertoires.

Most γδ T-cell clones that respond to EBV express a Vγ9JγP CDR3. Two hundred thousand γδ T cells were isolated from PBMCs before and after stimulation with Akata, an EBV-transformed B cell line. (A) Pie charts show the γ-chain CDR3 repertoires of unstimulated γδ T cells and EBV-stimulated γδ T cells from 1 representative group 1 donor and 1 representative group 2 donor. CDR3γ V9-JP sequences are represented in blue and CDR3γ V9-J2 sequences are represented in red. The gray zone represents the combination of all other unique CDR3s within the CDR3γ repertoires. (B) Pie charts show the distribution of the 20 most abundant γ-chain CDR3s of unstimulated γδ T cells and EBV-stimulated γδ T cells, from 3 representative group 1 donors and 3 representative group 2 donors. The public CDR3s ALWEVQELGKKIKV (dark blue) and ALWEVRELGKKIKV (light blue) are shown. For each donor, the 5 most dominant CDR3s expressed by unstimulated γδ T cells are shown in different shades of color and, when required, in dark blue or light blue for ALWEVQELGKKIKV and ALWEVRELGKKIKV, respectively. To track these same CDR3s in the EBV-stimulated γδ T-cell repertoires, we depicted them in similar colors. The other 15 CDR3s are colored in light green and the remaining unique CDR3s are shaded in gray. (C) Kullback-Leibler sequence logos show the residue preferences for the dominant 14-mer CDR3γ sequences from EBV-stimulated γδ T cells of group 1 (N = 10 CDR3γ) and group 2 (N = 10 CDR3γ) donors. Populations of γδ T cells were isolated from EBV-stimulated PBMCs. Each logo consists of amino acid stacks, 1 for each position. The stack height indicates the sequence conservation; the residue height indicates its relative frequency. Residue colors correspond to biochemical properties: acidic (red), basic (blue), hydrophobic (black), polar (green). Logos were generated using WebLogo.28  (D) The charts illustrate the distribution of the 20 most abundant γ-chain CDR3s among unstimulated γδ T cells and EBV-stimulated γδ T cells from a group 1 donor: donor 1. Populations of γδ T cells were isolated from unstimulated PBMCs and EBV-stimulated PBMCs. CDR3γ V9-JP sequences are represented by different shades of blue and CDR3γ V9-J2 are represented by different shades of red. The gray zone indicates a combination of all the other unique CDR3s within the CDR3γ repertoires.

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