Avadomide and its combination PD-L1/PD-1 blockade induces an inflammatory T-cell secretome and enhances T-cell motility. (A) Representative scanned dot blot from a cytokine array hybridized with culture supernatants from treated patient T cells. (B) Quantification of the secretome dot blots of conditioned media from treated patient T cells. Data show the mean pixel intensity (representative of 3 patients). (C) Luminex FLEXMAP 3D cytokine bead array data shown as fold change (compared with untreated cells) for the indicated cytokines in treated patient T-cell culture supernatants (n = 10). (D) Representative migratory tracks of individual patient T cells are shown in the far-left plots. Bar charts show speed of T-cell migration following the anti-PD-1-based (left) and anti-PD-L1-based (right) immunotherapy treatments indicated (n = 6 patients, minimum of 15 cells per patient sample treatment). (E) Illustration of the chemotaxis assay of autologous T cells toward conditioned media (lower well) derived from avadomide treated patient T cells and quantification of autologous T-cell migration toward treated T cell-conditioned media (n = 8). CXCL10 (lower well) included as an assay control. Data are presented as fold change relative to medium alone control. (F) Bar chart showing speed of patient T-cell migration (n = 3) following the drug treatments indicated and in the presence of anti-CXCR3 Abs were indicated. *P < .05; **P < .01; using a Freidman test with Dunn’s multiple comparisons test (C) and a repeated measures 1-way ANOVA with Tukey's multiple comparisons test (D-F). Data presented as mean ± SEM.