Panel A: Experimental schematic and flow-cytometry gating identify total live and yellow fluorescent protein-positive splenic cells populations. Panel B: UMAP plots classify B-cell subpopulations, while stacked bars summarize relative fractions percentages. Panel C: Gene-set enrichment analysis plot highlights antigen-processing and antigen-presentation pathway enrichment in conditional-knockout pre-germinal-center B cells. Panel D: Flow-cytometry histograms compare major histocompatibility complex class II expression on yellow fluorescent protein-positive and negative B cells. Panel E: UMAP identify cluster of differentiation 4-positive T-cell subsets and relative fractions. Panel F: Volcano plot displays differentially expressed genes in T-follicular-helper cells comparing conditional-knockout and control samples. Panel G: Experimental schematic illustrates bone-marrow transplantation into irradiated wild-type and major histocompatibility complex class II-deficient recipients. Panel H: Representative photographs show spleens, peripheral lymph nodes, and mesenteric lymph nodes from transplanted chimeric recipient mice. Panel I: Experimental schematic outlines mixed bone-marrow transplantation into irradiated recombination-activating gene 2-deficient recipient mice. Panel J: Representative photographs display lymphoid organs from recipient mice receiving different donor bone-marrow combinations after transplantation.
Eif3e deficiency initiates a feedforward loop of T–B interaction and lymphocyte activation. (A) Experimental outline for scRNA-seq analysis of total and YFP+ cells in the spleen of 2-mo-old control and cKO mice. (B) UMAP clustering analysis of B cell subpopulations in control and cKO YFP+ samples. A stacked bar plot summarizes fractions of each B cell subpopulation. (C) GSEA of top DEGs in cKO vs. control YFP+ pre-GCB cells. (D) Flow cytometry histogram showing MHC II intensity on YFP− and YFP+ B cells from 12-wk-old control and cKO mice. (E) UMAP clustering analysis of subpopulations of CD4+ T cells in control and cKO total samples. A stacked bar plot summarizes fractions of each CD4+ T cell subpopulation. (F) Volcano plot showing DEGs in the TFH cell subpopulations of cKO versus control total cells. The cutoff was set at P <0.05 and absolute log2FC >0.25. (G–J) Functional analysis of CD4+ T cells and B cell expression of MHC II in the development of lymphoproliferation disorder in cKO mice. (G and I) Experimental outline for generation of bone marrow chimeras without CD4+ T cells (G) or B cell–specific deletion of MHC II (I). (H and J) Representative photos of spleens, pLNs, and mLNs from indicated chimeras at 3 mo after reconstitution. Scale bars, 1 cm. scRNA-seq data (B, C, E, and F) from two independent biological replicates were merged for analysis. Data (D, H, and J) are representative of two independent experiments. pLNs, peripheral lymph nodes; mLNs, mesenteric lymph nodes.