Class II artificial antigen presenting cells for cancer immunotherapy

项目摘要 过继细胞疗法(ACT)已成为一种越来越有吸引力的治疗方法。 实体肿瘤患者由于其令人印象深刻的应答率；也就是说，成本和 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 这种疗法的可及性。在可伸缩、无细胞的领域取得了重大进展 扩增肿瘤特异性CD8 T细胞的技术。然而，没有类似的脱细胞 尽管存在压倒性的临床前和临床阶段，CD4T细胞的扩增平台仍然存在 有证据表明，CD4T细胞是抗肿瘤免疫反应的中枢，并能增强CD8- 以治疗为基础。拟议项目的目标是研究一种新的 用于功能性多克隆扩增的II类人工抗原提呈细胞(AAPC) 用于ACT的内源性肿瘤特异性CD4T细胞及其与内源性T细胞的潜在协同作用 CD8T细胞。该平台由50 nm顺磁性葡聚糖纳米颗粒组成 与第二类主要组织相容性复合体结合的蛋白及其共刺激作用 分子，将允许我们1)丰富和扩增罕见的小鼠和人类肿瘤特异性CD4T 2)促进树突状细胞(DC)非依赖性T细胞的帮助。在……里面 反过来，II类AAPC将首次允许我们同时监测抗肿瘤疗效和T ACT与多克隆CD_4或CD_4/CD_8 T细胞联合作用的细胞受体动力学 为了实现这些目标，该项目将分三个阶段进行。首先，我们将 AAPC扩增的多克隆CD_4体外功能及体内抗肿瘤作用的研究 与B16-OVA和B16-F10结合的针对异源抗原和自身抗原的T细胞、OVA和Trp1 黑色素瘤模型。第二，我们将应用模块化的人类II类AAPC， 能够通过负载的人类白细胞抗原分子扩增一系列抗原特异性的CD4T细胞 利用凝血酶可裂解肽扩增功能性肿瘤抗原NY-ESO-1特异性CD4T 来自HLADR1和DP4供者的细胞。最后，我们将结合第一类和第二类AAPC 联合应用研究DC非依赖性体内外抗肿瘤协同作用的技术 用B16-OVA和B16-F10培养抗原特异性的小鼠和人的CD4和CD8 小鼠T细胞和人SK-Mel-37(A2+/NY-ESO-1+)黑色素瘤细胞株对人T细胞的作用 细胞。如果成功，这项提议将提供一种新的脱细胞方法来处理多克隆的CD4或 结合了CD4和CD8 ACT，将提供对T细胞帮助的替代机制的洞察， 对体外扩增CD8 T细胞具有潜在的临床影响。

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.