Processed Antigen Characterization by Mass Spectrometry

描述（由申请人提供）：人体细胞通过降解细胞蛋白并在细胞表面呈递相关的I类MHC蛋白片段，将其健康状况传达给免疫系统。经过适当教育的细胞毒性t淋巴细胞（CTL） （CD8+ t细胞）与细胞表面的I类MHC分子结合，对呈现的肽进行采样，并杀死那些由于病毒、细菌和寄生虫感染、组织移植和细胞转化（癌症）而表达新肽的细胞。目前，晚期转移性黑色素瘤最有效的治疗方法是CD8+ t细胞的过继细胞疗法（ACT）。在这种方法中，肿瘤浸润淋巴细胞（TIL）从手术切除的肿瘤中分离出来，在体外扩增，然后通过化疗和全身照射联合消融患者的免疫系统后重新引入患者体内。改进这项技术的努力正在进行中，包括转染患者CD8+ t细胞（在扩增之前），使其具有特异性肿瘤相关的I类MHC肽的高亲和力受体。有了这个额外的步骤，ACT应该有可能用于治疗任何人类肿瘤。缺少的是MHC I类肽；(a)在癌症细胞与正常细胞上的差异显示，(b)在同一癌症患者的大队列中共享，(c)在多种肿瘤类型中共享，(d)来源于基因不能突变或删除而不影响肿瘤生存的蛋白质，(e)不能在胸腺或淋巴结的MHC分子上显示以触发活性CD8+ t细胞的删除。本文建议研究开发新的质谱仪器和方法，用于鉴定与癌细胞细胞转化特征相关的信号转导通路失调，从而满足上述标准的I类MHC磷酸肽。这项研究将使过继t细胞疗法扩展到许多其他癌症，包括急性髓性白血病、慢性淋巴细胞白血病、胰腺癌、结直肠癌和肝细胞腺癌和肾癌成为可能。公共卫生相关性：本文建议研究开发新的质谱仪器和方法，用于鉴定源自与细胞转化相关的信号转导通路失调的I类MHC磷酸肽。这项研究将使过继t细胞疗法从黑色素瘤扩展到其他癌症，包括急性髓性白血病、慢性淋巴细胞白血病、胰腺癌、结直肠癌、肝细胞腺癌和肾癌成为可能

DESCRIPTION (provided by applicant): Cells in the human body communicate their health status to the immune system by degrading cellular proteins and presenting fragments of each on the cell surface in association class I MHC proteins. Appropriately educated, cytotoxic T-lymphocytes (CTL) (CD8+ T-cells) bind to the class I MHC molecules on the cell surface, sample the peptides being presented and kill those cells that express new peptides as a result of viral, bacterial and parasitic infection, tissue transplantation and cellular transformation (cancer). Presently, the most effective treatment for late stage metastatic melanoma involves adoptive cell therapy (ACT) with CD8+ T-cells. In this approach, tumor-infiltrating lymphocytes (TIL) are isolated from surgically removed tumor, expanded ex vivo and then re-introduced to the patient after ablation of his or her immune system by a combination of chemotherapy and total body irradiation. Efforts to improve this technology are in progress and involve transfecting patient CD8+ T-cells (prior to expansion) with high affinity receptors for specific tumor associated class I MHC peptides. With this additional step, it should be possible to use ACT to treat any human tumor. What is lacking are MHC class I peptides that are; (a) differentially displayed on cancer vs normal cells, (b) shared by large cohorts of patients with the same cancer (c) shared by multiple tumor types, (d) derived from proteins whose genes cannot be mutated or deleted without compromising tumor survival, and (e) not available for display on MHC molecules in the thymus or lymph nodes to trigger deletion of reactive CD8+ T-cells. Proposed here is research to develop new mass spectrometry instrumentation and methods for the identification of class I MHC phosphopeptides that are derived from dysregulated signal transduction pathways associated with cellular transformation characteristic of cancer cells and thus satisfy the above criteria. This research should make it possible to extend adoptive T-cell therapy to a number of other cancers including acute myeloid leukemia, chronic lymphocytic leukemia, pancreatic-,colorectal- and heptocellullar- adenocarcinoma and renal cancer. PUBLIC HEALTHE RELEVANCE: Proposed here is research to develop new mass spectrometry instrumentation and methods for the identification of class I MHC phosphopeptides that are derived from dysregulated signal transduction pathways associated with cellular transformation. This research should make it possible to extend adoptive Tcell therapy beyond melanoma to other cancers including acute myeloid leukemia, chronic lymphocytic leukemia, pancreatic-, colorectal-, and heptocellullar adenocarcinoma and renal cancer