Multiplexed single cell cytokine secretion assay

描述(由申请人提供)：免疫细胞使用信号来协调它们对感染性病原体的攻击，或者在危险过去后降低它们的攻击级别。他们使用的信号是称为细胞因子的蛋白质，大约有25种。在免疫紊乱中，细胞因子可以发出信号或控制对身体的自我定向攻击，因此，在自身免疫条件下，细胞因子受到了极大的关注。然而，查询单个细胞以了解细胞因子表达的方法很少，到目前为止，还没有方法在从皮肤和其他炎症组织中分离出来的小样本免疫细胞上做到这一点。 Trellis测量单细胞细胞因子表达的方法是捕捉单细胞在平面塑料表面上产生的细胞因子。这些细胞的“足迹”然后可以通过多色纳米珠子探测，每种珠子类型都连接到不同的抗细胞因子抗体。这种方法已经揭示了免疫细胞的语言并不像有时所描绘的那么简单。正常样本中免疫细胞的高度多样性在疾病中可能会大大减少，或者正常组织中罕见的细胞类型可能会显着放大。这是一个可以用Trellis的CellSpot-CK技术探索的问题。其他问题涉及基于对疾病表现基础的细胞学特征的直接探索而进行的疾病分期。这些信息在选择这些慢性进展性疾病的治疗方案以及监测患者反应方面应该非常有用。 在将CellSpot-CK介绍给世界各地的细胞免疫学家之前，Trellis将需要使用于读取细胞斑点的仪器更加坚固、更容易使用和更快。这是第二阶段提案的一个具体目标。其他具体目标与确定健康和疾病中免疫细胞的细胞因子语言有关。 CellSpot-CK的成功开发将为研究人员提供一种新的工具，用于了解免疫系统在许多慢性疾病中的作用，如湿疹、牛皮癣、关节炎和哮喘。它还将提供一个工具，以了解免疫细胞如何对正在开发的治疗这些疾病的药物做出反应。从长远来看，它可能提供一种重要的临床“治疗”分析(基于诊断程序的治疗选择)。

DESCRIPTION (provided by applicant): Immune cells use signals to coordinate their attack on an infectious agent or to downgrade their attack after danger has past. The signals that they use are proteins called cytokines, of which there are about 25. During immunological disorders, cytokines can either signal or control a self-directed attack on the body, and therefore much attention has been paid to cytokines during autoimmune conditions. Yet there are few ways to query individual cells for cytokine expression and until now there have been no ways to do so on the small samples of immune cells isolated from skin and other inflamed tissues. Trellis' approach to measuring single cell cytokine expression is to capture the cytokines produced by single cells while they lie on a flat plastic surface. These cell "footprints" can then be probed by multihued nanoscale beads, each bead type linked to a different anti-cytokine antibody. This method has already revealed that the language of immune cells is not as simple as it is sometimes portrayed. The high level of diversity of immune cells in normal samples may be much diminished in disease or cell types that are rare in normal tissue may become significantly amplified. This is one question that can be explored with Trellis' CellSpot-CK technology. Other questions relate to staging disease based on direct exploration of the cellular profiles that underlie disease manifestation. Such information should be very useful in selecting treatment regimens for these chronic, progressive diseases as well as for monitoring patient responses. Before introducing CellSpot-CK to cellular immunologists throughout the world, Trellis will need to make the instrument used to read CellSpots more robust, easier to use, and faster. This is one Specific Aim in this Phase II proposal. The other Specific Aims relate to characterizing the cytokine language of immune cells in both health and disease. The successful development of CellSpot-CK will make available to researchers a new tool for understanding the role of the immune system in many chronic conditions like eczema, psoriasis, arthritis, and asthma. It will also provide a tool to learn how immune cells respond to drugs under development as cures for these diseases. Longer term, it may provide an important clinical "theranostic" assay (therapy selection based on a diagnostic procedure).