Figure 6 from Chemotherapy Drives Tertiary Lymphoid Structures That Correlate with ICI-Responsive TCF1<sup>+</sup>CD8<sup>+</sup> T Cells in Metastatic Ovarian Cancer
Authors: Lanickova Tereza , Hensler Michal , Kasikova Lenka , Vosahlikova Sarka , Angelidou Artemis , Pasulka Josef , Griebler Hannah , Drozenova Jana , Mojzisova Katerina , Vankerckhoven Ann , Laco Jan , Ryska Ales , Dundr Pavel , Kocian Roman , Cibula David , Brtnicky Tomas , Skapa Petr , Jacob Francis , Kovar Marek , Praznovec Ivan , McNeish Iain A. , Halaska Michal J. , Rob Lukas , Coosemans An , Orsulic Sandra , Galluzzi Lorenzo , Spisek Radek , Fucikova Jitka 
Published: January 2025

<p>The clinical relevance of combined chemotherapy and immunotherapy in mouse models of TMB<sup>Lo</sup> and TMB<sup>Hi</sup> ovarian cancer. <b>A</b> and <b>B,</b> Bar plots showing single-nucleotide variants positions (SNVs) (<b>A</b>) and the somatic mutations prevalence (mutations per megabase) (<b>B</b>) in ID8 (<i>n</i> = 3) and <i>Brca1</i><sup>−/−</sup><i>Trp53</i><sup>−/−</sup>/<i>Myc</i>/<i>Hras</i> SO1 (<i>n</i> = 3) C57BL/6 syngeneic mouse ovarian cancer cell lines. Mean and SEM are shown. Statistical significance was calculated by multiple <i>t</i> test. <i>P</i> values are indicated. <b>C,</b> Experimental design for the analysis of TLS aggregates development and efficacy of combined chemotherapy and aPD1 and/or aTIM-3 therapy in TMB<sup>Lo</sup> ID8 and TMB<sup>Hi</sup> SO1 experimental syngeneic mouse models. <b>D</b> and <b>E,</b> Representative immunostaining for CD4, CD8, CD20, and CD21 (<b>D</b>) and a box plot showing density of TLS aggregates within chemo-naïve (<i>n</i> = 5) and treated TMB<sup>Hi</sup> SO1 (<i>n</i> = 9) ovarian tumors (<b>E</b>). Scale bars, 100 µm and 2.5 mm. Box plots: lower quartile, median, upper quartile; whiskers, minimum, maximum. Statistical significance was calculated by two-sided Mann–Whitney test. <i>P</i> values are indicated. <b>F–H,</b> Representative dot plot (<b>F</b>) and flow cytometry analyses for percentages of CD62L<sup>+</sup>CD44<sup>+</sup> central memory (CM) and CD62L<sup>−</sup>CD44<sup>−</sup> terminally differentiated CD8<sup>+</sup> T cells (TEMRA) (<b>G</b>) and TCF1<sup>+</sup>PD1<sup>+</sup>CD8<sup>+</sup> and TIM-3<sup>+</sup>PD1<sup>+</sup>CD8<sup>+</sup> T cells (<b>H</b>) in tumor samples of the TMB<sup>Hi</sup> SO1 experimental model in the presence or absence of carboplatin and taxane chemotherapy (NACT). Box plots: lower quartile, median, upper quartile; whiskers, minimum, maximum. Statistical significance was calculated by two-sided Mann–Whitney test. <i>P</i> values are indicated. <b>I</b> and <b>J,</b> Overall survival (OS) of TMB<sup>Hi</sup> SO1 experimental model (<b>I</b>) and flow cytometry analyses for percentage of TCF1<sup>+</sup>PD1<sup>+</sup>CD8<sup>+</sup> T cells after NACT, aPD1, aTIM-3 and combined therapy (<b>J</b>). Survival curves were estimated by the Kaplan–Meier method, and differences between groups were evaluated using log-rank test. Box plots: lower quartile, median, upper quartile; whiskers, minimum, maximum. Statistical significance was calculated by two-sided Mann–Whitney test. <i>P</i> values are indicated. <b>K</b> and <b>L,</b> Representative immunostaining for CD4, CD8, CD20, and CD21 (<b>K</b>) and a box plot showing density of TLS aggregates within chemo-naïve (<i>n</i> = 8) and treated TMB<sup>Lo</sup> ID8 (<i>n</i> = 8) (<b>L</b>) ovarian tumors. Scale bars, 100 µm and 2.5 mm. Box plots: lower quartile, median, upper quartile; whiskers, minimum, maximum. Statistical significance was calculated by two-sided Mann–Whitney test. <i>P</i> values are indicated. <b>M,</b> Flow cytometry analyses for TCF1<sup>+</sup>PD1<sup>+</sup>CD8<sup>+</sup> and TIM-3<sup>+</sup>PD1<sup>+</sup>CD8<sup>+</sup> T cells in tumor samples of the TMB<sup>Lo</sup> ID8 experimental model in the presence or absence of carboplatin and taxane chemotherapy (NACT). Box plots: lower quartile, median, upper quartile; whiskers, minimum, maximum. Statistical significance was calculated by two-sided Mann–Whitney test. <i>P</i> values are indicated. <b>N,</b> Overall survival (OS) of TMB<sup>Lo</sup> ID8 experimental model after NACT, anti-PD1, and combined therapy. Survival curves were estimated by the Kaplan–Meier method, and differences between groups were evaluated using log-rank test. <i>P</i> values are indicated. (Panel <b>C</b> created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>.)</p>