PO.TB10.05 · 肿瘤生物学
多重免疫荧光揭示卵巢癌中与年龄和免疫治疗反应相关的区室特异性免疫细胞相关性
Multiplex immunofluorescence reveals compartment-specific immune cell correlations associated with age and immunotherapy response in ovarian carcinoma
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摘要 Abstract
中文摘要
背景 卵巢癌是一种侵袭性恶性肿瘤，通常在晚期确诊，对免疫检查点抑制剂的反应有限。越来越多的证据表明，年龄相关的免疫改变以及肿瘤微环境内的空间异质性会影响免疫治疗结局。本研究探讨组织区室间的免疫细胞组织是否与免疫治疗的临床反应相关。
方法 纳入二十名接受免疫治疗的患者。检索FFPE肿瘤蜡块，使用多重免疫荧光panel（DAPI、PD-L1、CD4、CD8、CD20、FoxP3、PanCK）对切片进行染色。感兴趣区域由病理学家标注，并使用inForm和PhenoChart中的监督学习进行分析。跨组织区室量化免疫细胞表型，并使用RStudio、Excel和GraphPad Prism进行相关性分析。
结果 老年患者的免疫治疗结局显著改善，包括更长的无进展生存期和有效免疫治疗持续时间，证实了治疗反应中存在年龄相关的优势。多重免疫荧光揭示了应答者与非应答者之间由组织区室化驱动的不同免疫相关网络。在免疫治疗应答者中，基质区域显示FoxP3⁺细胞与PanCK⁺肿瘤细胞之间呈正相关，而肿瘤区域则显示CD8⁺细胞与PanCK⁺细胞之间呈负相关。全组织分析显示，应答者中FoxP3⁺与CD8⁺细胞之间存在强相关性。相反，非应答者表现出不同的免疫细胞组织模式，其特征为基质中CD4⁺与CD8⁺ T细胞之间以及肿瘤区室中CD20⁺ B细胞与PanCK⁺细胞之间的强相关性。空间邻近性热图表明，应答者在肿瘤和基质区域内的多个半径范围内，CD8⁺、CD4⁺和CD20⁺细胞的局部聚集程度更高，而非应答者表现出有限的空间聚集，与"免疫冷"肿瘤一致。
结论 本研究鉴定了卵巢癌中与免疫治疗反应相关的独特的、区室特异性的免疫细胞关系，突显了年龄作为免疫结构的重要调节因素。应答者（以老年患者为主）表现出协调的基质FoxP3⁺相互作用和肿瘤CD8⁺参与，而非应答者（以年轻患者为主）则表现出基质CD4⁺/CD8⁺聚集和肿瘤CD20⁺关联，这是冷免疫表型的特征。这些发现提示，免疫细胞相关性（而非绝对密度）可能作为预测免疫治疗反应的信息性空间生物标志物。
查看英文原文 English abstract
Background Ovarian carcinoma, an aggressive malignancy typically diagnosed at advanced stages, has shown limited response to immune checkpoint inhibitors. Increasing evidence suggests that age-related immune alterations and spatial heterogeneity within the tumor microenvironment influence immunotherapy outcomes. This study investigates whether immune cell organization across tissue compartments correlates with clinical response to immunotherapy.
Method Twenty patients treated with immunotherapy were identified. FFPE tumor blocks were retrieved, and slides were stained using a multiplex immunofluorescence panel (DAPI, PD-L1, CD4, CD8, CD20, FoxP3, PanCK). Regions of interest were annotated by a pathologist and analyzed using supervised learning in inForm and PhenoChart. Immune cell phenotypes were quantified across tissue compartments, and correlation analyses were performed using RStudio, Excel, and GraphPad Prism.
Result Older patients demonstrated significantly improved immunotherapy outcomes, including longer progression-free survival and duration of effective immunotherapy, confirming an age-associated advantage in therapeutic response. Multiplex immunofluorescence revealed distinct immune correlation networks between responders and non-responders, driven by tissue compartmentalization. Among immunotherapy responders, stromal regions showed a positive correlation between FoxP3⁺ and PanCK⁺ tumor cells, whereas tumoral regions exhibited a negative correlation between CD8⁺ cells and PanCK⁺ cells. Whole-tissue analysis revealed a strong correlation between FoxP3⁺ and CD8⁺ cells in responders. In contrast, non-responders displayed a different pattern of immune cell organization, characterized by strong correlations between CD4⁺ and CD8⁺ T cells in the stroma and between CD20⁺ B cells and PanCK⁺ cells in the tumor compartment. Spatial proximity heatmaps demonstrated that responders had higher local clustering of CD8⁺, CD4⁺, and CD20⁺ cells across multiple radii within both tumor and stromal regions, while non-responders exhibited limited spatial clustering consistent with “immunologically cold” tumors.
Conclusion This study identifies distinct, compartment-specific immune cell relationships associated with immunotherapy response in ovarian carcinoma highlights age as a significant modifier of immune architecture. Responders, predominantly older patients, exhibited coordinated stromal FoxP3⁺ interactions and tumoral CD8⁺ engagement, whereas non-responders, predominantly younger patients, showed stromal CD4⁺/CD8⁺ clustering and tumoral CD20⁺ associations characteristic of cold immune phenotypes. These findings suggest that immune cell correlations, rather than absolute densities, may serve as informative spatial biomarkers for predicting immunotherapy response.
利益披露 Disclosure
K. Kwock, None.