Figure 5 from Pulsed Electric Field Ablation Reprograms Tumor Immunity and Stimulates Germinal Center Formation in Tertiary Lymphoid Structures in Patients with Non–Small Cell Lung Cancer
Year: 2026
Type: other
Abstract: Ablation-induced changes in the TME facilitate TLS and GC formation and functional changes suggestive of GC activity. A, B-cell populations in the TME, as assessed by scRNA-seq. Bar plots represent the mean percent of each cell population shown relative to the total number of leukocytes in each sample (±SD). P values were calculated using the propeller method. B, Heatmap showing the mean log2 fold change in the expression of the indicated genes relative to expression levels in naïve B cells as measured by scRNA-seq. C, Representative brightfield IHC images of TLS within an ablated tumor after resection. Example images of TLS_GC are shown, with higher magnification images collected from the areas denoted with dashed boxes. The various cell types within a GC are shown in the high magnification panel, including mature B cells within the GC that exhibit the characteristic morphology of proliferating centroblasts (light blue arrow) and centrocytes (dark green arrow). GCs were also found to contain tingible body macrophages (dark blue arrow) and DCs (black arrow). D, Multispectral immunofluorescent images from the same TLS_GC in C. The first panel shows all biomarkers together [4′,6-diamidino-2-phenylindole (nuclei), CD20, CD4, AID, Ki67, CD21, PNAd], followed by CD20 alone to show B cells, CD4 alone to show T cells, AID and Ki67 together to show proliferating cells and class-switching recombination, and CD21 and PNAd to show FDC networks within the GC and proximal HEV. Scale bars represent 100 μm. E–G, Scatter plots of cell frequencies in all TLS_GC compared with TLS in the ablation group (n = 34), demonstrating the percent of CD20:Ki67 dual-positive B cells (E), CD4:Ki67 dual-positive T cells (F), and CD20:AID dual-positive B cells (G). Blue lines represent the median, and P values were calculated using the Mann–Whitney test. H, Bar plot demonstrating the percentage of ablated tumors (n = 34) containing at least one TLS_GC from the brightfield image assessment, compared with the preablation biopsies (n = 17), as well as to the nonablated tumors from the expanded control group (n = 29; P < 0.0001 across the three groups, with significant between-group differences as specified in the plot). P values were calculated using Fisher's exact test. We additionally compared the proportion of TLS_GC-positive tumors before and after ablation in patients with paired pre- and postablation samples (n = 17) and found a significant difference between the two time points (P < 0.005, calculated using McNemar's χ2 test, data not shown). I, Bar plot demonstrating the percentage of ablated tumors containing at least one TLS_GC from the tissue sections stained with the multispectral immunofluorescent biomarker panel (control, n = 15; preablation, n = 9; postablation, n = 34; P < 0.0001 across the three groups, with significant between-group differences as specified in the plot). P values were calculated using Fisher's exact test. mIHC, multispectral immunohistochemical; TFH, T follicular helper. [Created in BioRender. Pastori, C. (2026) https://BioRender.com/79qnysx.].
Authors: Marcelo Jimenez, Beryl A. Hatton, Alicia Moreno-Gonzalez, Jeffrey S. Iding, Javier Flandes, Erik H.F.M. van der Heijden, Calvin S.H. Ng, Carlos Prieto, Cristina Teodosio, Juan A. Flores-Montero, Marta Rodríguez-González, Oderay Mabel Cedeño Diaz, Shoko Vos, Rainbow W.H. Lau, Medard E. Kaiza, William H. Moore, Alberto Orfao, Tullia C. Bruno
Language: English
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W7204959456