Association of immune characteristics determined by flow cytometry and T cell receptor (TCR) sequencing with patient prognosis.

(A) Unsupervised hierarchical clustering of samples (columns) in terms of the profile of immune marker abundance determined by flow cytometry. Cell color in the heatmap shows normalized abundance of immune markers, from low (blue) to high (yellow). Pink and white boxes show values of TCR characteristics; pink indicates an above-average value and white indicates a below-average value for each characteristic: productive rearrangement, productive clonality, productive entropy, top 10 productive frequency, and top 100 productive frequency. Cluster A (red square) is enriched with patients with a poor or worst prognosis; no patients in this cluster had an excellent prognosis. (B) Kaplan–Meier curves of overall survival in patients grouped in Cluster A and in the remaining clusters by hierarchical clustering. (C) CD8+ T cell subpopulations were significantly decreased in ascites from patients with an excellent prognosis compared with ascites from patients with a poor prognosis. The y-axis shows the percentage of CD3+ cells (log 2 ). CD8/Treg is the ratio of the percentage of live CD8+ lymphocytes to the percentage of live CD4+FoxP3+ lymphocytes. Each box plot represents the median and the twenty-fifth and seventy-fifth percentiles. Only significant associations are shown. (D) Significant differences in ascites TCR characteristics between patients with excellent and poor prognosis. The y-axis shows the percentage of CD3+ cells (log 2 ). (E) Scatterplots of CD8/Treg with productive entropy. (F) Scatterplots of CD8/Treg with top 100 productive frequency. Good/excellent prognosis was associated with less abundant CD8/Treg, increased productive entropy, and decreased top 100 productive frequency.