尽管许多卵巢肿瘤具有免疫原性，但使用免疫检查点疗法治疗并未大幅改善卵巢癌的生存率。为了推进群体水平的卵巢肿瘤免疫微环境研究，重要的是了解与使用多重免疫荧光（mIF）测定实验室制作的组织芯片（TMA）上的免疫细胞测量方法相关的方法问题。 在两个前瞻性队列中，我们收集了486例卵巢肿瘤的甲醛固定石蜡包埋标本，并制作了七个TMA。我们使用两个mIF面板在TMA上测量了T细胞，包括若干亚群体和免疫检查点标记。我们使用Spearman相关，Fisher精确检验和多变量调整贝塔二项式模型评价与TMA肿瘤核心中的免疫细胞测量相关的因素。 肿瘤内免疫标记物之间的核心相关性在0.52-0.72之间，较常见的标记物（如CD3+，CD3+CD8+）具有更高的相关性。所有区域、肿瘤区域和基质区域的免疫细胞标记物之间的相关性都很高（范围为0.69-0.97）。在多变量调整模型中，清细胞和黏液样癌与II型肿瘤相比，T细胞阳性的几率较低（Odds ratios[OR]0.13-0.48），对于若干亚群体，年龄较大的组织（样本年龄>30与≤10年，OR 0.11-0.32）阳性的T细胞几率较低。 总的来说，使用mIF测量的免疫标记物的核心间相关性高，支持使用TMA研究卵巢肿瘤的免疫浸润，尽管非常老的样本可能会降低抗原性。未来的流行病学研究应评估组织类型对肿瘤免疫反应的差异，并确定可能改变肿瘤免疫微环境的可改变因素。

Despite the immunogenic nature of many ovarian tumors, treatment with immune checkpoint therapies has not led to substantial improvements in ovarian cancer survival. To advance population-level research on the ovarian tumor immune microenvironment, it is critical to understand methodological issues related to measurement of immune cells on tissue microarrays (TMAs) using multiplex immunofluorescence (mIF) assays.In two prospective cohorts, we collected formalin-fixed paraffin-embedded ovarian tumors from 486 cases and created seven TMAs. We measured T cells, including several sub-populations, and immune checkpoint markers on the TMAs using two mIF panels. We used Spearman correlations, Fisher's exact tests, and multivariable-adjusted beta-binomial models to evaluate factors related to immune cell measurements in TMA tumor cores.Between-core correlations of intratumoral immune markers ranged from 0.52-0.72, with more common markers (e.g., CD3+, CD3+CD8+) having higher correlations. Correlations of immune cell markers between the whole core, tumor area, and stromal area were high (range 0.69-0.97). In multivariable-adjusted models, odds of T cell positivity were lower in clear cell and mucinous vs. type II tumors (odds ratios [OR] 0.13-0.48) and, for several sub-populations, were lower in older tissue (sample age >30 vs. ≤10 years, OR 0.11-0.32).Overall, high correlations between cores for immune markers measured via mIF support the use of TMAs in studying ovarian tumor immune infiltration, although very old samples may have reduced antigenicity.Future epidemiologic studies should evaluate differences in the tumor immune response by histotype and identify modifiable factors that may alter the tumor immune microenvironment.