Figure 5
Distributions of read numbers of CMV-NLV-specific TCRβ clonotypes. Cells from the HLA-A2-positive and CMV-seropositive donors V001 and V004 that were detected using the HLA-A2-NLV tetramer probe were sorted and isolated from unstimulated PBMCs (a,d) and PBMCs stimulated once (b,e) or twice (c,f) with the NLV peptide. The sorted cells were then subjected to NGS. Each marker in the plots shows one unique read of CMV NLV-specific TCRβ. The TCRβ clonotypes are ordered according to their frequencies on the x-axis. The y-axis depicts the read number of the clonotypes. Shared CMV NLV-specific TCRs and unshared CMV NLV-specific TCRs are shown as red circles and red dots, respectively. Orange thick circle represents one of nine dominant TCRs in V001 (TCR ID; 001-17, 001-48, 001-41) and V004 (TCR ID; 004-66, 004-22, 004-63, 004-30, 004-28 and 004-71) (Fig. 2a). The sizes of the circles represent the number of sharing (two to five) among the five donors. The largest circle indicates that the corresponding TCR clonotype were shared by the five donors. The Wilcoxon rank-sum test was performed to assess whether public CMV NLV TCRs occupy a higher rank of the read numbers compared with the private CMV NLV TCRs. Wilcoxon rank-sum test and Kendall’s tau test were performed. All tests were statistically significant (P < 0.05). The ranks (the first to tenth) of %read of CMV NLV-specific TCRβ clonotypes are shown in the tables below the plots.
