Purpose: The Cancer Immune Monitoring and Analysis Centers – Cancer Immunologic Data Commons (CIMAC-CIDC) Network is supported by the NCI to identify biomarkers of response to cancer immunotherapies across clinical trials using state-of-the-art assays. A primary platform for CIMAC-CIDC studies is cytometry by time of flight (CyTOF), performed at all CIMAC laboratories. To ensure the ability to generate comparable CyTOF data across labs, a multistep cross-site harmonization effort was undertaken. Experimental Design: We first harmonized standard operating procedures (SOPs) across the CIMAC sites. Because of a new acquisition protocol comparing original narrow- or new wide-bore injector introduced by the vendor (Fluidigm), we also tested this protocol across sites before finalizing the harmonized SOP. We then performed cross-site assay harmonization experiments using five shared cryopreserved and one lyophilized internal control peripheral blood mononuclear cell (PBMC) with a shared lyophilized antibody cocktail consisting of 14 isotype-tagged antibodies previously validated, plus additional liquid antibodies. These reagents and samples were distributed to the CIMAC sites and the data were centrally analyzed by manual gating and automated methods (Astrolabe). Results: Average coefficients of variation (CV) across sites for each cell population were reported and compared with a previous multisite CyTOF study. We reached an intersite CV of under 20% for most cell subsets, very similar to a previously published study. Conclusions: These results establish the ability to reproduce CyTOF data across sites in multicenter clinical trials, and also highlight the importance of quality control procedures, such as the use of spike-in control samples, for tracking variability in this assay.

中文翻译：

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免疫分析质谱流式细胞仪检测协调：CIMAC-CIDC 的多中心经验


目的：癌症免疫监测和分析中心 – 癌症免疫数据共享 (CIMAC-CIDC) 网络得到 NCI 的支持，利用最先进的检测方法在临床试验中识别癌症免疫疗法反应的生物标志物。 CIMAC-CIDC 研究的主要平台是飞行时间细胞计数 (CyTOF)，在所有 CIMAC 实验室进行。为了确保能够跨实验室生成可比较的 CyTOF 数据，采取了多步骤的跨站点协调工作。实验设计：我们首先协调了 CIMAC 站点的标准操作程序 (SOP)。由于新的采集协议比较了供应商 (Fluidigm) 引入的原始窄孔或新型宽孔注射器，因此我们还在最终确定统一的 SOP 之前跨站点测试了该协议。然后，我们使用 5 个共享的冷冻保存的和一个冻干的内部对照外周血单核细胞 (PBMC) 以及由 14 种先前验证的同种型标记抗体和其他液体抗体组成的共享冻干抗体混合物进行了跨位点测定协调实验。这些试剂和样品被分发到 CIMAC 站点，并通过手动门控和自动化方法（Astrolabe）对数据进行集中分析。结果：报告了每个细胞群跨位点的平均变异系数 (CV)，并与之前的多位点 CyTOF 研究进行了比较。对于大多数细胞亚群，我们的位点间 CV 都低于 20%，这与之前发表的研究非常相似。 结论：这些结果确立了在多中心临床试验中跨站点复制 CyTOF 数据的能力，并且还强调了质量控制程序的重要性，例如使用掺入对照样品来跟踪该测定中的变异性。