Fig. 3: TLS frequency and maturation impact the T cell phenotype in HGSOC.

A Spatial co-localization of TLSs with MS4A1, PDCD1, TCF7 and HAVCR2 genes in TLS^Hi tumor sample from Study Cohort 1. B Representative image of immunofluorescence of CD68, CD8, PD-L1, FoxP3, TCF1, CD57, PanCK, PD1, CD4, CD20, GZMB and TIM3 staining (immunofluorescence panel 2). Scale bar, 2 mm, 10 µm and 200 µm. C Density of TCF1⁺PD1⁺CD8⁺ T cells, TIM3⁺PD1⁺CD8⁺ and TIM3⁺PD1⁻CD8⁺ T cells within complete tumor microenvironment (TME, including TLSs), early TLS (eTLS) and mature TLS (mTLS) of 19 HGSOC tumor samples (Study cohort 1, n = 19). Mean and SEM are shown. Statistical significance was calculated by two-sided Wilcoxon matched-pairs signed rank test. p values are indicated. D Density of PD1⁺CD8⁺, TIM3⁺PD1⁺CD8⁺ and TCF1⁺PD1⁺CD8⁺ T cells within tumor stroma and tumor core of 19 HGSOC patients (Study Cohort 1) separated into 3 clusters (CL1, no TLS development, n = 8; CL2, only eTLS development, n = 6; CL3, both eTLS and mTLS development, n = 5, as previously determined in Fig. 2) Box plots: lower quartile, median, upper quartile; whiskers, minimum, maximum. Statistical significance was calculated by two-sided Wilcoxon matched-pairs signed rank test. p values are indicated. Representative images (E) and violin plot (F) showing the frequency of eTLS and mTLS in 209 HGSOC and 31 non-small cell lung carcinoma (NSCLC) (Supplementary Table. 6). Statistical significance was calculated by two-sided Mann–Whitney test. p values are indicated. G, H Density of TIM3⁺PD1⁺CD8⁺, TCF1⁺PD1⁺CD8⁺ T cells and GZMB⁺CD8⁺ T cells within complete tumor microenvironment (TME, including TLS), eTLS and mTLS of HGSOC (Study cohort 1) and NSCLC samples (Study cohort 5) as determined by immunofluorescence. Number of patients involved in respective analyses are indicated. Box plots: lower quartile, median, upper quartile; whiskers, minimum, maximum. Statistical significance was calculated by two-sided Mann–Whitney test. p values are indicated. I Density of TCF1⁺PD1⁺CD8⁺ T cells, TIM3⁺PD1⁺CD8⁺ and TIM3⁺PD1^-CD8⁺ T cells within complete tumor microenvironment (TME, including TLSs), early TLS (eTLS) and mature TLS (mTLS) in 17 NSCLC tumor samples (Study cohort 5). Mean and SEM are shown. Statistical significance was calculated by two-sided Wilcoxon matched-pairs signed rank test. p values are indicated. J Correlation between frequency of mTLS and TIM3⁺PD1⁺CD8⁺ and TCF1⁺PD1⁺CD8⁺ T cells in TME of 17 HGSOC and 17 NSCLC patients as determined by multispectral immunofluorescence. p values are indicated. R, Pearson correlation coefficient. Source data are provided as a Source Data file.