NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, USA.
Nat Med. 2011 Jun;17(6):738-43. doi: 10.1038/nm.2375. Epub 2011 May 22.
Cellular immunity has an inherent high level of functional heterogeneity. Capturing the full spectrum of these functions requires analysis of large numbers of effector molecules from single cells. We report a microfluidic platform designed for highly multiplexed (more than ten proteins), reliable, sample-efficient (∼1 × 10(4) cells) and quantitative measurements of secreted proteins from single cells. We validated the platform by assessment of multiple inflammatory cytokines from lipopolysaccharide (LPS)-stimulated human macrophages and comparison to standard immunotechnologies. We applied the platform toward the ex vivo quantification of T cell polyfunctional diversity via the simultaneous measurement of a dozen effector molecules secreted from tumor antigen-specific cytotoxic T lymphocytes (CTLs) that were actively responding to tumor and compared against a cohort of healthy donor controls. We observed profound, yet focused, functional heterogeneity in active tumor antigen-specific CTLs, with the major functional phenotypes quantitatively identified. The platform represents a new and informative tool for immune monitoring and clinical assessment.
细胞免疫具有固有高水平的功能异质性。要捕捉这些功能的全貌，需要从单个细胞中分析大量的效应分子。我们报告了一种微流控平台，该平台设计用于高度多重化（超过十种蛋白质）、可靠、高效（约 1×10(4)个细胞）和定量测量来自单个细胞的分泌蛋白。我们通过评估脂多糖 (LPS) 刺激的人巨噬细胞中的多种炎症细胞因子，并与标准免疫技术进行比较，验证了该平台。我们通过同时测量从肿瘤抗原特异性细胞毒性 T 淋巴细胞 (CTL) 中分泌的十几种效应分子，应用该平台对肿瘤抗原特异性 CTL 的多功能性多样性进行了离体定量，这些 CTL 正在积极响应肿瘤，并与一组健康供体对照进行了比较。我们观察到活跃的肿瘤抗原特异性 CTL 中存在深刻而集中的功能异质性，并定量鉴定了主要的功能表型。该平台代表了免疫监测和临床评估的一种新的、信息丰富的工具。
