Processed Antigen Characterization by Mass Spectrometry

描述（由申请人提供）：人体内的细胞通过降解细胞蛋白质并将每种蛋白质的片段以I类MHC蛋白的形式呈递到细胞表面，将其健康状态传达给免疫系统。经过适当训练的细胞毒性T淋巴细胞（CTL）（CD 8 + T细胞）与细胞表面上的I类MHC分子结合，对呈递的肽进行取样，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的最有效治疗涉及使用CD 8 + 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，离体扩增，然后在通过化疗和全身照射的组合消融他或她的免疫系统后重新引入患者。改进该技术的努力正在进行中，并且涉及用特异性肿瘤相关I类MHC肽的高亲和力受体来扩增患者CD 8 + T细胞（在扩增之前）。有了这个额外的步骤，它应该可以使用ACT治疗任何人类肿瘤。缺乏的是MHC I类肽;（a）在癌细胞与正常细胞上差异显示，（B）由患有相同癌症的大量患者共享，（c）由多种肿瘤类型共享，（d）源自其基因不能突变或缺失而不损害肿瘤存活的蛋白质，和（e）不能在胸腺或淋巴结中的MHC分子上展示以触发反应性CD 8 + T细胞的缺失。本文提出的研究是开发用于鉴定I类MHC磷酸肽的新的质谱仪器和方法，所述I类MHC磷酸肽来源于与癌细胞的细胞转化特征相关的失调的信号转导途径，因此满足上述标准。这项研究将使过继性T细胞治疗扩展到许多其他癌症成为可能，包括急性髓性白血病、慢性淋巴细胞白血病、胰腺癌、结肠直肠癌和肝细胞腺癌以及肾癌。公共卫生相关性：这里提出的是研究开发新的质谱仪器和方法，用于鉴定I类MHC磷酸肽，这些磷酸肽来自与细胞转化相关的失调信号转导途径。这项研究将使过继性T细胞治疗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，结肠直肠癌，肝细胞腺癌和肾癌。

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