在遗传和表型上完全相同的免疫细胞种群在免疫功能和蛋白质分泌模式方面可能高度异质性。基于微流控芯片的单细胞高度多重分泌组蛋白质组学技术可以在不同的细胞和分子层面上对免疫应答的细胞异质性进行表征。越来越多的证据表明，在单个细胞水平上同时产生2种以上蛋白质的多功能T细胞是关键的效应细胞，对抗病原体和肿瘤的强大和持久的细胞免疫有贡献。这种功能性蛋白质组学技术提供了广泛的细胞功能评估，并可以唯一地确定从活体细胞中具有细胞因子标记的高度多功能细胞亚群。这种高维单细胞分析提供了对功能异质性的深入分析，并有助于确定预测性生物标志物和潜在相关因子，这对于免疫治疗产品设计的优化和个性化免疫治疗的发展至关重要，以实现更好的临床结果。 ©2023 Wiley-VCH GmbH。

Genetically and phenotypically identical immune cell populations can be highly heterogenous in terms of their immune functions and protein secretion profiles. The microfluidic chip-based single-cell highly multiplexed secretome proteomics enables characterization of cellular heterogeneity of immune responses at different cellular and molecular layers. Increasing evidence has demonstrated that polyfunctional T cells that simultaneously produce 2+ proteins per cell at the single-cell level are key effector cells that contribute to the development of potent and durable cellular immunity against pathogens and cancers. The functional proteomic technology offers a wide spectrum of cellular function assessment and can uniquely define highly polyfunctional cell subsets with cytokine signatures from live individual cells. This high-dimensional single-cell analysis provides deep dissection into functional heterogeneity and helps identify predictive biomarkers and potential correlates that are crucial for immunotherapeutic product design optimization and personalized immunotherapy development to achieve better clinical outcomes.© 2023 Wiley-VCH GmbH.