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. 2024 Mar 8;5(3):101448. doi: 10.1016/j.xcrm.2024.101448

Figure 3.

Figure 3

Characteristics and dynamics of B cells in each group of early-stage LUAD

(A) Tumor sections from patients with LUAD stained with anti-CD4 monoclonal antibody (mAb; green), anti-CD8 mAb (red), anti-CD19 mAb (white), and DAPI (4′,6-diamidino-2-phenylindole; blue). Scale bars, 50 μm, 500 μm, 1 mm, and 2 mm. nLung (n = 11), AIS (n = 6), MIA (n = 10), and IAC (n = 10).

(B) Boxplot showing quantification of CD19+/CD4+/CD8+ cells in total cells, respectively; data from multiplex immunostaining analysis. Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, two-sided Mann-Whitney U test.

(C) 2D projection of subclustered of B cells from all nLung and LUAD samples, showing the formation of six main clusters. A total of 6,475 cells were annotated and are marked by color code.

(D) 2D projection of a selected set of B cell marker genes over the metacell model.

(E) Bar plots showing the B cell-type composition within the nLung and patients with AIS, MIA, and IAC in each group. Cell types are colored the same as in (C).

(F) B cells of nLung and patients with AIS, MIA, and IAC are shown with contour lines indicating the density of projected cells.

(G) Dot plots showing percentages of B cell subpopulation within nLung and patients with AIS, MIA, and IAC. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, two-sided Mann-Whitney U test. Error bars indicate mean ± SEM.

(H) Density plots showing the proportions of tumor samples and nLung tissues that express high levels of CD79A and CD27 and CD27 and CD83, respectively, in TCGA stage I LUAD and Genotype-Tissue Expression (GTEx) nLung data. Expression level was measured by log2 (count + 1).

(I) Tumor sections from patients with LUAD stained with anti-CD83 mAb (white), anti-CD138 mAb (yellow), anti-CD27(green), anti-CD19 mAb (red), and DAPI (blue). Scale bars, 20 and 200 μm.

(J) Fraction of proliferating B cells per metacell. Circle size reflects the percentages of proliferating cells in the metacell using the 2D projection as in (C).

(K) Tumor sections from patients with LUAD stained with anti-KI67 mAb (red), anti-CD19 mAb (white), and DAPI (blue). Scale bars, 50 μm.