Mass cytometric analysis of T_TEcells in MGUS and NDMM patients. (A) Biaxial contour plots gated for CD8⁺T cells show distribution of CD69⁺T_TE and CD69⁻T_TE cells in NDMM and MGUS patients. Regions occupied by CD69⁺T_TE and CD69⁻T_TE cells are indicated. Numbers indicate proportions of CD69⁺T_TE and CD69⁻T_TE cells within BM-CD8⁺T and PB-CD8⁺T cells of NDMM and MGUS patients. (B) tSNE plots show distribution of CD69⁻T_TE and CD69⁺T_TE cells within pooled BM-T_TE and PB-T_TE cells of NDMM patients (n = 5, shown in Figure 4A) and MGUS patients (n = 3). The area occupied by CD69⁺T_TE cells is indicated by the red dotted circle. Bars (median with scatter plots) show proportions of CD69⁻T_TE and CD69⁺T_TE cells within T_TE (C) and CD8⁺T (D) cells in the BM and PB of NDMM (n = 8) and MGUS (n = 4) patients. *P < .05. (E) Graph shows proportion of CD69⁺ cells within the T_N, T_M, and T_TE compartments in the BM of MGUS (n = 4) and NDMM (n = 8) patients. T_N, T_M, and T_TE cells from the same patient are connected by lines. ***P < .001; Friedman test with Dunn’s multiple comparators. (F) Heat map showing the phenotype of the 25 MCs defined by FlowSOM analysis of T_TE cells. MC5, MC6, MC9, and MC11 contain CD69⁺T_TE cells, and CD69⁻T_TE cells are distributed within the remaining 21 MCs. The intensity of the color in each cell indicates the median signal intensity for an individual marker (row) in an individual MC (column). (G) Pie charts show the contribution of CD69⁺T_TE cells assigned to MC5, MC6, MC9, and MC11 to the BM-T_TE cells in NDMM (n = 5) and MGUS (n = 3) patients. Numbers indicate total CD69⁺T_TE cells within MC5, MC6, MC9, and MC11 in each patient, expressed as a percentage of total BM-T_TE cells.