<p>Supplementary Tables</p>

<div>AbstractPurpose:<p>In chronic lymphocytic leukemia (CLL), the T-cell receptor (TCR) repertoire is skewed and tumor-derived antigens are hypothesized as drivers of oligoclonal expansion. Ibrutinib, a standard treatment for CLL, inhibits not only Bruton tyrosine kinase of the B-cell receptor signaling pathway, but also IL2-inducible kinase of the TCR signaling pathway. T-cell polarization and activation are affected by ibrutinib, but it is unknown whether T cells contribute to clinical response.</p>Experimental Design:<p>High-throughput TCRβ sequencing was performed in 77 longitudinal samples from 26 patients with CLL treated with ibrutinib. TCRβ usage in CD4⁺ and CD8⁺ T cells and granzyme B expression were assessed by flow cytometric analysis. Antitumor cytotoxicity of T cells expanded with autologous CLL cells or with antigen-independent anti-CD3/CD28/CD137 beads was tested.</p>Results:<p>The clonality of the TCR repertoire increased at the time of response. With extended treatment, TCR clonality remained stable in patients with sustained remission and decreased in patients with disease progression. Expanded clonotypes were rarely shared between patients, indicating specificity for private antigens. Flow cytometry demonstrated a predominance of CD8⁺ cells among expanded clonotypes. Importantly, bulk T cells from responding patients were cytotoxic against autologous CLL cells <i>in vitro</i> and selective depletion of major expanded clonotypes reduced CLL cell killing.</p>Conclusions:<p>In patients with CLL, established T-cell responses directed against tumor are suppressed by disease and reactivated by ibrutinib.</p><p><i>See related commentary by Zent, p. 4465</i></p></div>

<p>(A) Frequency of predominant clonotypes as detected by NGS and of the corresponding CD8+ TRBV families as measured by flow cytometry. Grey bars indicate TRBV families that did not have commercially available fluorescent-labelled antibodies. (B) Pie chart of the most expanded clonotypes and expression of granzyme B (GrB). CD8+ GrB+ clonotypes comprising >2% of the repertoire at the time of response are shown in color.-</p>

<p>Expansion of T cells under different conditions. Total CD3+, CD8+, CD4+ counts at Day 0 and Day 7 of expansion in the presence of anti-CD3/CD28/CD137 beads or autologous CLL cells.</p>

<p>Supplementary Tables</p>