PD-1T TILs as precision biomarker in NSCLC 49 2 ◀ Figure 5. PD-1T high samples contain a higher density of PD-1T TILs inside and outside TLS. (A) Percentage of pretreatment samples containing tertiary lymphoid structures (TLS) (n=30), TLS and/or lymphoid aggregates (LA) (referred as TLS+LA, n=46) and CD20+ B cells (n=86) in the remaining cohort (n=91). (B) Top: Example of a CD20-CD3 IHC double staining with the black square showing CD20+ B cells (in yellow) and CD3+ T cells (in purple) localizing in a TLS. Arrows indicate LA. Bottom: Example of a consecutive PD-1 IHC staining with the black square showing PD-1T TILs inside TLS. (C) Digital markup showing the spatial distribution of CD20+ B cells (in green) and CD3+ T cells (in light-blue), and digital markup of PD-1T TILs (in red) and all other cells (in grey). (D) PD-1T TILs per mm2 inside and outside TLS in resected samples (n=23), and of total tumor area for resected samples with no TLS (n=9). Medians, interquartile ranges and minimum/maximum shown in boxplots, **** P<0.0001 by Mann-Whitney test. (E) TLS per mm2 in pretreatment samples from patients with DC 12m (n=20) and PD (n=71). Shown is the mean, P=0.054 by Mann-Whitney test. (F) The predictive value of PD-1T TILs, TLS, TLS+LA and CD20-positive area per mm2 for DC 12m (n=91, note that this cohort is smaller due to the availability of FFPE material). Shown are AUCs with 95% CI interval. (G) TLS per mm2 in PD-1T low (<90 per mm2 of total tumor area) (n=10) and PD-1T high (≥90 per mm2 of total tumor area) (n=13) resected samples. Medians, interquartile ranges and minimum/maximum shown in boxplots, P=0.18 by Mann-Whitney test. (H) PD-1T TILs per mm2 inside TLS and (I) outside TLS in PD-1T low (n=10) and PD-1T high (n=13) resected samples. Medians, interquartile ranges and minimum/maximum shown in boxplots, ** P<0.01 and **** P<0.0001 by Mann-Whitney test.
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