微流体细胞分离和细胞因子传感平台及对於免疫细胞检测的应用

Diseases of the immune system can cause fatal health problems by disrupting the defensive mechanism of the immune system; and therefore immunoresponses of the patients should be well characterized for developing the proper treatment strategies in time. Considering that leukocytes play a key role in both innate and adaptive immune responses, examination of such immune cells extracted from a patient can provide important information about the systematic immune responses. However, promising prognosis and diagnosis on the immune responses are yet very challenging, mainly hindered by the multiplexed, heterogeneous and dynamic biochemical responses of immune cells extracted from the patient’s blood. The challenge stems are not only from the limitations of conventional methods, but also caused by the intrinsic heterogeneity and complexity of the immune cell types in the community. For this reason, quantitative and dynamic analyses of the cytokine secretion profiles of each the immune cell type are required for precise characterization of the ‘immunophenotype’ of patients...This proposal aims at developing a novel automated microfluidic ‘immunophenotyping’ platform for efficient and high-throughput isolation, stimulation, and quantitative characterization of individual immune cell types at defined locations in the device. We expect to achieve computer-controlled operation procedures for the system in order to achieve continuous monitoring of the cytokine secretion profiles of different leukocyte cell types. Detecting transient profiles of the cytokine secretions will provide a signature that quantitatively characterizes patients’ immunophenotypes, which in turn lead to better prognosis and diagnosis and allow for personalized medications.

免疫系统疾病以通过破坏免疫系统的防御机制导致致命的健康问题。然而，为免疫系统应答提供有效的预测和诊断仍是一个巨大的挑战，原因主要来自于病人血液中免疫细胞多重、异质和动态生化反应的阻碍（如细胞因子的分泌）。我們需要对每种类型的免疫细胞分泌细胞分子进行定量和动态的分析以精确表征患者的“免疫表型”。..本申请旨在开发一种新型的自动微流体“免疫表型”平台，可以在此平台中固定的区域内进行高效高通量的分离、刺激和对单一类型免疫细胞的定量表征。该平台将在细胞表面蛋白表达和细胞因子分泌基础上实现芯片内免疫细胞捕获/分离、以及以酶联免疫吸附测定法为基础使用包被有藻红蛋白标记检测抗体的微球进行细胞因子分泌的大规模平行实时监控。鉴于其潜在的快速、微量和多重检测功能使之可以适用于检测全血样本，我们坚信提出的微流体平台将会为免疫系统疾病的监测和诊断提供一个有效的自动化医疗技术，有助于探索开发早期的临床治疗药物。

人体免疫缺陷 (HIV)、肺结核 (TB) 和疟疾等免疫疾病会破坏免疫系统的防御机制，从而导致致命的健康问题；因此，应充分表征患者的免疫反应，以便及时制定适当的治疗策略。考虑到白细胞（免疫细胞）在先天性和适应性免疫反应中都发挥着关键作用，对从患者身上提取的免疫细胞进行检查可以提供有关系统性免疫反应的重要信息。然而，有希望的免疫反应预后和诊断仍然非常具有挑战性，主要受到从患者血液中提取的免疫细胞的多重、异质和动态生化反应（例如细胞因子分泌）的阻碍。出于这个原因，需要对单个免疫细胞的细胞因子分泌谱进行定量和动态分析，以精确表征患者的“免疫表型”。.该提案旨在开发一种新型集成微流体“免疫表型”平台，用于在设备中的特定位置对单个免疫细胞进行高效和高通量的分离、刺激和定量表征。所提出的平台将实现片上免疫细胞捕获/分离，以及基于酶联免疫吸附测定的细胞因子分泌的大规模并行实时监测，使用涂有藻红蛋白偶联检测抗体的市售微珠。这种微流控平台只需要几微升的血液，可以通过手指刺破获得，用于实际的临床测试。检测细胞因子分泌物的瞬时特征将提供定量表征患者免疫表型的特征，这反过来会导致更好的诊断并允许个性化药物治疗。鉴于适用于全血样本的潜在快速、小体积、多重检测能力，我们坚信我们提出的微流控平台将诱导一种有效的免疫疾病监测和诊断自动化医疗技术，有助于早期临床治疗的药物搜索.