Mechanisms and consequences of antigen-dependent T cell homing for adoptive immunotherapies

项目摘要/摘要 具有激活和扩增的T或CAR T细胞的过继细胞疗法(ACT)可用于治疗感染或 肿瘤和带有T或CAR T调节细胞的ACT正在进行临床试验，以控制自身免疫和同种异体移植 拒绝。许多但不是所有患者都受益。ACT的成功取决于T细胞对相关组织的归巢 网站。正常循环中的T效应记忆或T调节细胞可以通过它们的同源反应进入组织 在TCR触发的过程中，抗原显示在局部微血管内皮细胞的表面 被共刺激调制的。我们假设人内皮细胞(ECs)的抗原提呈 会招募过继转移的体外扩增的T细胞或CAR T效应器和调节细胞到特定的 在由特定的EC共刺激物调节的过程中的外围组织位置。在具体目标1中，我们将 在我们利用流室中的内皮细胞单层开发的模型中，在体外测试这一假设 在活体条件下进行模拟。我们将扩展我们的体外ASSYS，以包括对TCR- 诱导的跨内皮细胞迁移在单细胞水平上对T细胞产生作用。就CAR T细胞而言， 我们将确定要整合到汽车中的最重要的共刺激分子受体分子基序 最优瞬变电磁法。我们还将确定人类ECS是否有能力通过 允许EC呈递抗原以影响癌症免疫治疗的抗原。最后，我们将使用一个模型 我们为研究过继转移的人类T细胞对合成微血管的反应而开发 由人类内皮细胞组装而成，允许对人类内皮细胞可以提供的信号进行基因操作。在AIM 2 我们将使用T和CAR T调节细胞进行类似的实验。圆满完成这些目标 将为扩大可能受益于ACT的患者范围提供重要信息。

Project Summary/Abstract Adoptive cell therapy (ACT) with activated and expanded T or CAR T cells may be used to treat infections or tumors and ACT with T or CAR T regulatory cells are in clinical trials to control autoimmunity and allograft rejection. Many but not all patients benefit. The success of ACT depends upon T cell homing to relevant tissue sites. Normal circulating T effector memory or T regulatory cells can enter a tissue in response to their cognate antigen being displayed on the surface of the local microvascular ECs in a process triggered by TCR and modulated by costimulation. We hypothesize that antigen presentation by human endothelial cells (ECs) will recruit adoptively transferred in vitro expanded T or CAR T effector and regulatory cells to specific peripheral tissue sites in a process modulated by specific EC co-stimulators. In specific aim 1, we will test this hypothesis in vitro in models we have developed using endothelial cell monolayers in flow chambers to model in vivo conditions. We will expand our in vitro assys to include an examination of the effects that TCR- induced transendothelial migration (TEM) has on the T cells at the single cell level. In the case of CAR T cells, we will determine the most important costimulator receptor molecule motifs to be incorporated into the CAR for optimal TEM. We will also determine if human ECs have the capacity to cross-present or be “cross-dressed” by antigens allowing EC presentation of antigen to influence cancer immunotherapy. Finally, we will use a model we developed for studying adoptively transferred human T cell responses to synthestic microvessels assembled from human ECs, allowing genetic manipulation of the signals human ECs can provide. In aim 2 we will conduct similar experiments using T and CAR T regulatory cells. Successful completion of these aims will provide important information for extending the range of patients who may benefit from ACT.