Autohistomagnetic Isolation of Tumor-reactive T-cells

自体肿瘤浸润淋巴细胞（TIL）的诱导性细胞治疗现在可以介导目标癌症 50%的转移性黑色素瘤患者的消退。虽然TIL治疗已经增加了 尽管近几十年来的改进，基本方法并没有发生重大变化， 免疫肿瘤学领域的重大进展。目前的TIL治疗方案集中于离体治疗。 使用高剂量白细胞介素-2在多周内扩增来自切除的肿瘤活检组织的TIL。这是一个时代- 消耗和昂贵的过程，限制了许多患者的资格，并减少了癌症的数量 能够提供这种救命疗法的治疗设施。影响临床的最重要因素 反应是最终TIL输注产物的抗肿瘤反应性，以及产生的TIL的总数 进行收养转移当前的方法在需要快速扩展TIL以维护 避免衰竭和诱导耐受的“年轻”表型，以及富集肿瘤- 反应性TIL。我们提出了一种称为自组织磁隔离（AHMI）的新技术， 肿瘤来源的抗原呈递复合物，以在活检后的前48小时内选择反应性TIL。的 这种新方法的基本原理是使用构象依赖性抗体进行免疫沉淀 将HLA或H2-Kb、β-2微球蛋白和同源肽的异源三聚体连接到磁珠上。所得 然后，构建体可以使用任何磁性分离平台从非反应性TIL中分离肿瘤反应性TIL 在阳性选择的大量TIL培养物中。这将大大减少产生TIL输注所需的时间 产品，将导致更有效和持久的TIL表型，降低成本，并增加抗肿瘤活性。 反应性我们建议完全优化AHMI，并在预处理中询问后续的TIL输注产品。 TIL治疗的临床鼠模型和来自转移性黑色素瘤患者的样品。完成 本文提出的研究将为小鼠和人类AHMI产生一个功能齐全的原型， 临床试验所需的基础工作。

Adoptive cell therapy of autologous tumor-infiltrating lymphocytes (TIL) can now mediate objective cancer regression in 50% of patients with metastatic melanoma. While TIL therapy has made incremental improvements over recent decades, the fundamental methodologies have not significantly changed despite major advances in the field of immuno-oncology. Current protocols for TIL therapy focus on the ex vivo expansion of TIL from resected tumor biopsies using high-dose interleukin-2 over many weeks. This is a time- consuming and expensive process that limits the eligibility of many patients and reduces the number of cancer treatment facilities capable of offering this life-saving therapy. The most important factors governing clinical response are the anti-tumor reactivity of the final TIL infusion product, and the total number of TIL generated for adoptive transfer. Current methodologies are torn between the need to expand TIL quickly to maintain a “young” phenotype that avoids exhaustion and the induction of tolerance, and the need to enrich for tumor- reactive TIL. We propose a novel technology called autohistomagnetic isolation (AHMI) that uses patient tumor-derived antigen presentation complex to select for reactive TIL within the first 48 hours after biopsy. The basic principle of this new methodology uses conformation-dependent antibodies to immunoprecipitate heterotrimers of HLA or H2-Kb, β-2 microglobulin, and cognate peptide onto magnetic beads. The resulting construct can then isolate tumor-reactive TIL from non-reactive TIL using any magnetic separation platform among positively selected bulk TIL cultures. This will greatly reduce the time needed to generate TIL infusion products, will result in a more potent and persistent TIL phenotype, reduce cost, and increase anti-tumor reactivity. We propose to fully optimize AHMI and interrogate the subsequent TIL infusion product in a pre- clinical murine model of TIL therapy, and in samples from patients with metastatic melanoma. The completion of the studies proposed here will produce a fully functional prototype for mouse and human AHMI and lay the groundwork necessary for clinical trials.