Differential enrichment of B-cell subsets associated with prognosis. (A) Analysis of the cell types composing the flow-sorted CD3⁺, CD14⁺, CD19⁺, and CD45⁺TN immune compartments according to gene signatures from azimuth.¹⁵ (B) Experimental workflow. Low-input RNA-seq was performed of individual cell populations sorted based on indicated markers from the bone marrow of 25 patients with NDMM. Cibersort was used to deconvolute cell type composition. (C-F) Bar graphs showing the composition of individual immune cell compartments based on deconvolution in patients with NDMM. Shown are percentages of cell types inferred by Cibersort for CD19⁺ cells (C), CD3⁺ cells (D), CD45⁺TN cells (E), and CD14⁺ cells (F). (G) Patients with early (TTPlow) and late (TTPhigh) relapse were divided by median TTP. (H) Heat map showing relative differences in cell type abundance between patients with poor vs good prognosis. (I-L) Dot plots showing the distribution of values for the indicated cell populations comparing TTPlow with TTPhigh. Significance was assessed using an unpaired t test; ∗P ≤ .05. (K) Log2 ratio of naive/memory B cells in TTPlow vs TTPhigh patients. Significance was assessed using an unpaired t test; ∗P ≤ .05. (M) Relative scores for a gene set for naive B cells in TTPlow vs TTPhigh patients. ∗P ≤ .05, by unpaired t test. Correlation of relative scores (bottom) for naive B-cell gene set vs TTP value (R = 0.56; P = .036). (N) Expression of canonical marker genes for naive and memory B cells in TTPlow vs TTPhigh patients. (O) Expression of marker genes distinguishing B cells in TTPlow and TTPhigh patients. (P) Heat map comparing activity of signaling pathways between TTPlow vs TTPhigh patients. ASDC, AXL⁺ dendritic cell; BaEoMa, basophil eosinophil mast progenitor; cDC1, CD141⁺ myeloid dendritic cell; cDC2, CD1c⁺ myeloid dendritic cell; CD8 effector.1, memory-like CD8⁺GZMK⁺ alpha-beta T cell; CD8 effector.2, late differentiated CD8⁺GZMB⁺ cytotoxic alpha-beta T cell; CD8 effector.3, NKT-like CD8⁺ alpha-beta T cell; ILC, innate lymphoid cell; MAIT, mucosal associated invariant T; Mono, monocyte; NKdim, CD56dim NK cell; NKbright, CD56bright NK cell.