Class II artificial antigen presenting cells for cancer immunotherapy

项目摘要 抗肿瘤细胞疗法（ACT）已经成为一种越来越有吸引力的治疗肿瘤的方法。 由于其令人印象深刻的反应率，实体瘤患者;也就是说， 目前用于扩增肿瘤特异性T细胞的细胞方法的复杂性有限， 这种治疗的可及性。在可扩展的、无细胞的 用于扩增肿瘤特异性CD 8 T细胞的技术。然而，没有类似的非细胞 存在用于扩增CD 4 T细胞的平台，尽管存在压倒性的临床前和临床应用。 证据表明，CD 4 T细胞是抗肿瘤免疫应答的核心，可以增加CD 8-T细胞的免疫应答。 基础疗法。该项目的目标是研究一种新的 II类人工抗原呈递细胞（aAPC）用于扩增功能性多克隆 内源性肿瘤特异性CD 4 T细胞用于ACT以及与内源性 CD 8 T细胞。该平台由50 nm顺磁性铁葡聚糖纳米颗粒组成， 与II类主要组织相容性复合物蛋白和共刺激分子偶联 分子，将允许我们1）富集和扩增罕见的小鼠和人类肿瘤特异性CD 4 T细胞， 细胞达到临床相关水平，以及2）促进树突状细胞（DC）非依赖性T细胞的帮助。在 反过来，II类aAPC将使我们第一次能够监测抗肿瘤疗效和T ACT与多克隆CD 4或组合的CD 4和CD 8 T细胞在小鼠中的细胞受体动力学。 为实现这些目标，该项目将分三个阶段进行。一是 研究aAPC扩增的多克隆CD 4的体外功能和体内抗肿瘤功效 对外源和自身抗原、OVA和Trp 1以及B16-OVA和B16-F10具有特异性的T细胞 黑色素瘤模型。其次，我们将应用模块化的人类II类aAPC， 能够通过负载的HLA分子扩增一系列抗原特异性CD 4 T细胞 用凝血酶可裂解肽扩增功能性肿瘤抗原NY-ESO-1特异性CD 4 T细胞 来自HLA DR 1和DP 4供体的细胞。最后，我们将联合收割机结合I类和II类aAPC 研究DC独立的体外和体内抗肿瘤协同作用的技术， 抗原特异性小鼠和人CD 4和CD 8培养物，使用B16-OVA和B16-F10用于 小鼠T细胞和人SK-MEL-37（A2+/NY-ESO-1+）黑素瘤细胞系用于人T细胞 细胞如果成功，该提案将提供一种新的多克隆CD 4或 结合CD 4和CD 8 ACT，并将提供对T细胞帮助的替代机制的见解， 具有离体CD 8 T细胞扩增的潜在临床分支。

Project Summary Adoptive cell therapy (ACT) has become an increasingly attractive method for treating patients with solid tumors due to its impressive response rate; that said, the costs and complexity of current cellular approaches for expansion of tumor-specific T cells have limited accessibility of this therapy. Significant progress has been made in scalable, acellular technologies for expanding tumor-specific CD8 T cells. However, no analogous acellular platforms for expansion of CD4 T cells exist, despite overwhelming preclinical and clinical evidence that CD4 T cells are central to antitumor immune responses and can augment CD8- based therapies. The goal of the proposed project is to investigate the application of a novel Class II artificial antigen presenting cell (aAPC) for expansion of functional, polyclonal endogenous tumor-specific CD4 T cells for ACT as well as potential synergies with endogenous CD8 T cells. The platform, which consists of a 50 nm paramagnetic iron dextran nanoparticle conjugated with Class II Major Histocompatibility Complex proteins and costimulatory molecules, will allow us 1) to enrich and expand rare murine and human tumor-specific CD4 T cells to clinically relevant levels, and 2) to facilitate dendritic cell (DC) independent T cell help. In turn, the Class II aAPC will allow us for the first time to monitor both the antitumor efficacy and T cell receptor dynamics of ACT with polyclonal CD4 or combined CD4 and CD8 T cells in mice. To accomplish these goals, the project will proceed in three phases. First, we will investigate the in vitro function and in vivo antitumor efficacy of aAPC-expanded polyclonal CD4 T cells specific to foreign and self-antigens, OVA and Trp1 with B16-OVA and B16-F10 melanoma models, respectively. Second, we will apply a modular human Class II aAPC, capable of expanding a range of antigen-specific CD4 T cells through HLA molecules loaded with thrombin-cleavable peptides, to expand functional tumor-antigen NY-ESO-1 specific CD4 T cells from HLA DR1 and DP4 donors. Finally, we will combine the Class I and Class II aAPC technologies to investigate DC independent in vitro and in vivo antitumor synergies of combined antigen-specific mouse and human CD4 and CD8 culture, using B16-OVA and B16-F10 for mouse T cells, and the human SK-MEL-37 (A2+/NY-ESO-1+) melanoma cell line for human T cells. If successful, this proposal will deliver a novel acellular approach for polyclonal CD4 or combined CD4 and CD8 ACT and will provide insight into alternative mechanisms of T cell help, with potential clinical ramifications for ex vivo CD8 T cell expansion.