Fig. 2: Tumor-infiltrating B cells and tertiary lymphoid structures are enriched in HPV+ PSCC. | Nature Communications

Fig. 2: Tumor-infiltrating B cells and tertiary lymphoid structures are enriched in HPV+ PSCC.

From: A B cell-IgA-epithelial axis enhances antitumor immunity and improves outcome in HPV-associated penile squamous cell carcinoma

Fig. 2: Tumor-infiltrating B cells and tertiary lymphoid structures are enriched in HPV+ PSCC.

A UMAP plot displaying 134,784 immune cells from scRNA-seq datasets of tumor specimens (HPV−, n = 13; HPV+, n = 10), including TIL-Bs (C1-C2), TIL-Ts (C3-C7), NK (C8) and myeloid-derived cells (C9-C12). B Relative abundance of each cluster in each PSCC patient. C, D Bar graph comparing the relative proportions of each cluster (C) and major immune types (D) between HPV− and HPV+ PSCC. P-values were indicated for C1: **P = 0.0063, C2: **P = 0.01, C6: **P = 0.0145 in (C) and TIL-Bs: ***P = 0.0007 in (D). E Gating strategy for FACS analysis in fresh PSCC specimens. F Histograms showing the relative proportions of total immune cells, TIL-Bs and TIL-Ts in fresh HPV- (n = 11) and HPV+ (n = 9) PSCC tumors (*P = 0.0151). G Overall survival curves of 85 patients with HPV- (n = 52) or HPV+ (n = 33) PSCC (*P = 0.0136). H Representative images of PSCC tumor sections with HE staining (left), double-plex IHC staining for CD20 and CD3 (middle), and TLSs (right), marked by black circles in both tumor and non-tumor regions. Scale bar: 2 mm (left and middle), 400 μm (upper right) and 150 μm (lower right). I Quantification of TLSs per tumor area in non-tumor and intra-tumor regions in PSCC (n = 37) (****P < 0.0001). J Number of TLSs per tumor area in HPV− (n = 52) and HPV+ (n = 33) PSCC (**P = 0.0029). K Quantification of the number of CD3+ and CD20+ within TLSs per tumor area in HPV− (n = 52) and HPV+ (n = 33) PSCC tumor sections by IHC stained (****P < 0.0001). L Representative immunofluorescence staining for CD3, CD20, CD38 and DAPI+ nuclei within TLSs in HPV- and HPV+ PSCC tumors. Scale bar: 200 μm. M Quantification of the number of CD3+, CD20+, and CD38+ within TLSs per tumor area in HPV− (n = 10) and HPV+ (n = 10) PSCC tumor sections by IF stained. (CD20: *P = 0.0420, CD38: *P = 0.0430). TIL-Bs:Tumor-infiltrating B cells. TIL-Ts: Tumor-infiltrating T cells.TLSs: tertiary lymphoid structures. A two-sided unpaired Student’s t test was used in (C), (D), (F), (J), (K) and (M). A two-sided paired Student’s t test for (I), and a two-sided log-rank (Mantel–Cox) test in (G). Data are presented as mean ± s.e.m. ns: not significant (P > 0.05). Source data are provided as a Source Data file.

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