KK10-specific clonotypes with high levels of Ag sensitivity express TRBV4-3/TRBJ1-3 TCRs. (A) Alignment of observed TCRβ amino acid sequences for TRBV4-3/TRBJ1-3 clonotypes; 3 public and 2 private sequences are shown. Amino acid residues that differ between clonotypes are highlighted in gray. The numbers of n additions required to produce each observed nucleotide sequence and the isolated clone assignments are indicated for each clonotype. (B) KK10-specific CD8⁺ T-cell clones (n = 35) isolated from 3 patients are grouped by TCRβ sequence, indicated by the box frames, and classified according to mean cognate Ag sensitivity (EC₅₀ for Cr⁵¹ release). Public clonotypes are highlighted with a star. (C) Classification of KK10-specific CD8⁺ T-cell clones according to cognate Ag sensitivity (EC₅₀ for Cr⁵¹ release) and TRBV/TRBJ usage. Each bar represents one clone: TRBV4-3/TRBJ1-3 clones are in black, TRBV4-3/non-TRBJ1-3 clones are in gray, and non-TRBV4-3 clones are in white. The clone reference is indicated on the x-axis, and the last letter of the code corresponds to the patient from whom the clone was obtained (A and F for patient 01.01 DOF, B and G for patient 04.064, and C and H for patient 11.007). Statistical analyses were conducted using the Mann-Whitney U test with Bonferroni correction for multiple comparisons (P < .0125 was considered significant). (D) Isolated KK10-specific CD8⁺ T-cell clones (n = 23) are grouped by TCRα sequence, indicated by the box frames, and classified according to mean cognate Ag sensitivity (EC₅₀ for Cr⁵¹ release). Public clonotypes are highlighted with a star. (E) Association between TRAV14 and TRBV4-3 gene usage in KK10-specific CD8⁺ T-cell clones. Each symbol represents one clone: ●, TRBV4-3/TRBJ1-3 clones; ○, TRBV4-3/non-TRBJ1-3 clones; and ♢, non-TRBV4-3 clones. The χ² test was used to assess statistical significance. **P < .01 and ***P < .001, respectively.