Autohistomagnetic Isolation of Tumor-reactive T-cells

自体肿瘤浸润性淋巴细胞(TIL)的过继细胞治疗现在可以介导目的肿瘤 50%的转移性黑色素瘤患者病情消退。虽然TIL治疗已经取得了进展 在近几十年的改进中，基本方法并没有显著改变，尽管 免疫肿瘤学领域的重大进展。目前的TIL治疗方案主要集中在体外 使用大剂量白介素2在数周内扩增切除肿瘤活检组织中的TIL。现在是时候了- 耗费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输注产品的抗肿瘤反应性，以及产生的TIL总数 用于领养转移。当前的方法在需要快速扩展TIL以保持 避免精疲力竭和诱导耐受性的“年轻”表型，以及需要对肿瘤进行充实- 反应性瓷砖。我们提出了一种名为自组织磁隔离(Ahmi)的新技术，它使用患者 肿瘤来源的抗原呈递复合体在活检后的第一个48小时内选择用于反应性TIL。这个 这一新方法的基本原理是使用构象依赖抗体进行免疫沉淀 将人类白细胞抗原或H2-KB、β-2微球蛋白和同源肽的杂三聚体修饰到磁珠上。由此产生的 然后，构建可以使用任何磁分离平台将肿瘤反应性TIL与非反应性TIL分离 在阳性选择的散装TIL培养中。这将大大减少产生TIL输注所需的时间 产品，将导致更强大和持久的TIL表型，降低成本，并增强抗肿瘤 反应性。我们建议全面优化Ahmi，并在预审中询问后续TIL输注产品 TIL治疗的临床小鼠模型，以及转移性黑色素瘤患者的样本。完成度 在这里提出的研究中，将产生一个小鼠和人类的全功能原型，并奠定 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.