Purpose: The Cancer Immune Monitoring and Analysis Centers – Cancer Immunologic Data Commons (CIMAC-CIDC) network supported by the NCI Cancer Moonshot initiative was established to provide correlative analyses for clinical trials in cancer immunotherapy, using state-of-the-art technology. Fundamental to this initiative is implementation of multiplex IHC assays to define the composition and distribution of immune infiltrates within tumors in the context of their potential role as biomarkers. A critical unanswered question involves the relative fidelity of such assays to reliably quantify tumor-associated immune cells across different platforms. Experimental Design: Three CIMAC sites compared across their laboratories: (i) image analysis algorithms, (ii) image acquisition platforms, (iii) multiplex staining protocols. Two distinct high-dimensional approaches were employed: multiplexed IHC consecutive staining on single slide (MICSSS) and multiplexed immunofluorescence (mIF). To eliminate variables potentially impacting assay performance, we completed a multistep harmonization process, first comparing assay performance using independent protocols followed by the integration of laboratory-specific protocols and finally, validating this harmonized approach in an independent set of tissues. Results: Data generated at the final validation step showed an intersite Spearman correlation coefficient ( r ) of ≥0.85 for each marker within and across tissue types, with an overall low average coefficient of variation ≤0.1. Conclusions: Our results support interchangeability of protocols and platforms to deliver robust, and comparable data using similar tissue specimens and confirm that CIMAC-CIDC analyses may therefore be used with confidence for statistical associations with clinical outcomes largely independent of site, antibody selection, protocol, and platform across different sites.

中文翻译：

---

多重组织成像协调：CIMAC-CIDC 免疫肿瘤生物标志物网络的多中心经验

目的：由 NCI 癌症登月计划支持的癌症免疫监测和分析中心 - 癌症免疫数据共享 (CIMAC-CIDC) 网络的建立是为了利用最先进的技术为癌症免疫治疗的临床试验提供相关分析。这一举措的基础是实施多重 IHC 检测，以定义肿瘤内免疫浸润物作为生物标志物的潜在作用的组成和分布。一个尚未解答的关键问题涉及此类测定的相对保真度，以可靠地量化不同平台上的肿瘤相关免疫细胞。实验设计：三个 CIMAC 站点在其实验室中进行比较：(i) 图像分析算法，(ii) 图像采集平台，(iii) 多重染色协议。采用两种不同的高维方法：单玻片上多重 IHC 连续染色 (MCSSS) 和多重免疫荧光 (mIF)。为了消除可能影响检测性能的变量，我们完成了一个多步骤的协调过程，首先使用独立的协议比较检测性能，然后整合实验室特定的协议，最后在一组独立的组织中验证这种协调的方法。结果：最终验证步骤生成的数据显示，组织类型内和组织类型之间每个标记的位点间 Spearman 相关系数 ( r ) ≥0.85，总体平均变异系数较低，≤0.1。结论：我们的结果支持方案和平台的互换性，以使用相似的组织标本提供可靠且可比较的数据，并确认 CIMAC-CIDC 分析因此可以放心地用于与临床结果的统计关联，很大程度上独立于部位、抗体选择、方案、和跨不同站点的平台。