Development of Microfluidic Nano-Liter Platform for Single Cell Dynamic Monitorin

单细胞动态监测微流控纳升平台的开发

• 批准号: 8413880
• 负责人: Tania Tali Konry
• 金额: $ 21.76万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-13 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8413880
• 关键词: Antibodies; Antigen-Antibody Reactions; Antigen-Presenting Cells; Antigens; Biochemical; Biological; Biological Assay; Cause of Death; Cell Communication; Cell secretion; Cell surface; Cells; Collaborations; Communication; Custom; DNA amplification; Data; Dendritic Cell Vaccine; Dendritic Cells; Detection; Development; Eligibility Determination; Encapsulated; Event; Future; Generations; Goals; Image; Immune; Immune response; Immune system; Immunotherapy; Individual; Interleukin-10; Interleukin-12; Label; Learning; Life; Ligands; Liquid substance; Malignant Neoplasms; Measurement; Membrane Proteins; Methods; Microfluidics; Microscopic; Microspheres; Modeling; Monitor; Optics; Pathway interactions; Pattern; Phase; Phenotype; Play; Population; Procedures; Process; Protocols documentation; Public Health; Reaction; Research; Role; Signal Transduction; Specificity; Surface; Synapses; System; T-Cell Activation; T-Lymphocyte; TNFRSF5 gene; TNFSF5 gene; Techniques; Technology; Therapeutic; Time; Up-Regulation; Vaccines; Visual; Work; base; cell type; cytokine; design; immune function; immunological synapse; immunological synapse formation; improved; lymph nodes; nanolitre; novel; novel strategies; polydimethylsiloxane; response; sensor; single molecule; technique development; tumor; tumor immunology; tumor progression

DESCRIPTION (provided by applicant): One of the central processes regulating immune function involves T cells establishing cell-cell contacts with antigen-presenting cells (APCs) such as dendritic cells (DCs) via a contact zone known as the immunological synapse (IS). The importance of this synapse is considerable since interaction of T cells with DCs in lymph nodes is one of the critical steps in the generation of antigen specific immune responses which activate the na¿ve T cells. Despite great technical advances in microscopic imaging, the development of techniques to dynamically monitor the cellular and the induced secretion patterns at the IS interaction on a single cell level to avoid ambiguities that arise due to heterogeneous responses in cell populations is lagging. It is the goal of the proposed work to develop a microfluidic droplt approach for dynamic monitoring of the biochemical events that transpire upon cell-cell communication in this key event of the immune response. In particular, the current proposal outlines the integration of a novel approach to monitor live cell surface expression changes as well as the actual immunological synapse (IS) formed between DCs and T cells. In addition to cell surface studies, the microfluidic droplet approach developed herein will also allow us to collect data on secreted molecules during IS formation utilizing microsphere sensors incorporated in the microfluidic reaction droplets. Thus, live cell secretion and cell surface changes will be monitored on a single cell level simultaneously in a distinct microenvironment, feat not possible using conventional techniques. PUBLIC HEALTH RELEVANCE: Cancer is a leading cause of death around the world. Due to biological complexities of cancer and tumor progression, we are learning that attacking cancer using classical therapeutic approaches has its limits. An immune-based, non-toxic approach that attacks the cancer as well as the supporting deficiencies that facilitate its progression woul be a great boon to the field. This project aims to improve public health by developing a successful tumor immunology model, which can be applied to study immunological reactions to the immunotherapy and eliminate long term cancer reappearance without disrupting normal immune function.

描述（由申请人提供）：调节免疫功能的中心过程之一涉及T细胞与抗原呈递细胞（APC）建立细胞-细胞接触， 作为树突状细胞（DC）通过称为免疫突触（IS）的接触区。这种突触的重要性是相当大的，因为淋巴结中T细胞与DC的相互作用是产生抗原特异性免疫应答的关键步骤之一，抗原特异性免疫应答激活幼稚T细胞。尽管在显微镜成像技术的巨大进步，技术的发展，以动态监测细胞和诱导的分泌模式在单细胞水平上的IS相互作用，以避免由于在细胞群体中的异质性反应而产生的歧义是滞后的。所提出的工作的目标是开发一种微流体液滴方法，用于动态监测在免疫应答的这一关键事件中细胞-细胞通信时发生的生化事件。特别是，目前的建议概述了一种新的方法来监测活细胞表面表达的变化，以及实际的免疫突触（IS）之间形成的DC和T细胞的整合。除了细胞表面研究之外，本文开发的微流体液滴方法还将允许我们利用并入微流体反应液滴中的微球传感器在IS形成期间收集关于分泌分子的数据。因此，活细胞分泌和细胞表面变化将在不同的微环境中在单个细胞水平上同时监测，这是使用常规技术不可能的。 公共卫生相关性：癌症是世界各地的主要死亡原因。由于癌症和肿瘤进展的生物学复杂性，我们正在了解使用经典治疗方法攻击癌症有其局限性。一种基于免疫的无毒方法可以攻击癌症以及促进其进展的支持缺陷，这对该领域来说将是一个巨大的布恩。该项目旨在通过开发一个成功的肿瘤免疫学模型来改善公众健康，该模型可用于研究免疫治疗的免疫反应，并在不破坏正常免疫功能的情况下消除长期癌症复发。