Figure 6.

Abundance of LAG-3^hiPD-1^hi memory CD4⁺ T cells in peripheral blood of CCA patients was correlated with response to ABL501

(A–C) FlowSOM analysis of immune cell subsets in the peripheral blood of relapsed (n = 10) or non-relapsed (n = 9) CCA patients. (A) viSNE plot shows immune cell subset clusters to identify FlowSOM clustering. (B) Bar graph shows the frequencies of each cluster in two groups (relapse versus non-relapse). (C) Heatmap shows the MFI of each cluster. (D–F) CITRUS analysis of T cell signatures in the peripheral blood of relapsed (n = 9) or non-relapsed (n = 8) CCA patients is shown. (D) CITRUS plot shows clusters from CCA patients in two groups (relapse versus non-relapse). Red dots represent clusters that showed a different abundance with a statistical significance between groups. (E) Bar graphs show the relevance of abundance for the selected clusters (red dots) between groups. (F) Histogram plots show each marker expression by the selected clusters (red) over background (light blue). (G) Representative viSNE plots of CD4⁺ T cells in the peripheral blood of relapsed or non-relapsed CCA patients overlaid with the expression of naive, memory, co-inhibitory, or co-stimulatory markers. (H) Fold increase of IFN-γ secretion by CD3⁺ T cells from relapsed (n = 12) or non-relapsed (n = 9) CCA patients upon co-culture with A375-OKT3 in the presence of the indicated antibodies at a final concentration of 66.8 nM is shown. (I) Fold increase of IFN-γ secretion by peptide pool-treated peripheral blood CD8⁺ T cells obtained from relapsed (n = 11) or non-relapsed (n = 9) CCA patients in the presence of the indicated antibodies at a final concentration of 66.8 nM is shown. Statistical significance was determined by paired t tests with two-tailed analysis in (B) or one-way ANOVA with Holm-Sidak multiple comparisons in (H) and (I). ∗p < 0.05, ∗∗p < 0.005, ∗∗∗p < 0.001, ∗∗∗∗p < 0.001, and ns.