IDENTIFYING AND EXPLOITING POST-TRANSLATIONAL MODIFICATIONS IN ANTIGEN PRESENTATION USING NEXT GENERATION PROTEOMICS

使用下一代蛋白质组学识别和利用抗原呈递中的翻译后修饰

• 批准号: 2182109
• 金额: --
• 依托单位: Durham University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2182109
• 关键词: IDENTIFYING; EXPLOITING; POST; TRANSLATIONAL; MODIFICATIONS

BackgroundIn previous BBSRC-funded work being advanced through this new collaboration between Durham, Liverpool and Scancell, we have identified novel routes for the presentation of antigens by MHC class II molecules (MHCII) to the immune system. MHCII display antigens to helper T cells to control infections by bacteria and other pathogens, but also present modified self-antigens (e.g. neoantigens). The Benham group has found that some MHCII types e.g. HLA-DP4, assemble independently of CD74, a protein that normally directs the MHCII to the endosomal system. These HLA-DP4 molecules can stably bind self- peptides in other compartments e.g. the Endoplasmic Reticulum and autophagic vesicles. Scancell/Durrant (amongst others) have shown that post-translationally modified (e.g. citrullinated) peptides can access these MHCII, generating novel modified peptides that can break self-tolerance and cause an immune response. This mechanism has implications for the control of infectious disease in humans and livestock, for the development of anti-cancer vaccines and for bespoke therapies for autoimmunity. Aims, Hypothesis and Experimental ApproachWe hypothesise that neoantigens can be accessed by HLA-DP4 during unconventional intracellular trafficking. The overarching aim of this project is to characterise these novel antigens and establish the cell biology that drives their display at the cell surface. Supporting this aim, we will employ underpinning mass spectrometry capability to define MHC combinatorial modifications by native ms, map the immuno-proteome with quantitative ms and identify post-translationally modified MHC peptides in two systems (reconstituted antigen presenting cells and a humanised mouse melanoma model developed by the CASE partner). The student will also be trained in a wide range of laboratory methods including confocal microscopy, tissue culture, SDS-PAGE/western blotting, biochemical/cellular assays, molecular biology, bioinformatics and statistical computing (R). The student will be trained in presentation skills and report writing, encouraged to present their work at international meetings, and will participate in a regular journal club. Relevance to BBSRC strategyThe project is both curiosity-led and creative, being catalysed by new developments in technological capability. The project sits broadly within the remit for world class underpinning Biosciences and relates directly to a systems approaches to the Biosciences and to technology development for the Biosciences, through the strong focus on advancing quantitative and mechanistic proteomic technologies.

背景在BBSRC资助的先前通过Durham、利物浦和Sancell之间的这一新合作而推进的工作中，我们已经确定了MHC II类分子(MHCII)向免疫系统递送抗原的新途径。MHCII向辅助T细胞展示抗原以控制细菌和其他病原体的感染，但也提供修饰的自身抗原(例如新抗原)。Benham研究小组发现，一些MHCII类型，如HLA-DP4，独立于CD74组装，CD74是一种通常将MHCII导向内体系统的蛋白质。这些HLA-DP4分子可以稳定地结合其他隔室中的自体多肽，如内质网和自噬小泡。Sancell/Durant(等)已经证明，翻译后修饰(例如瓜氨酸)的多肽可以访问这些MHCII，产生新的修饰多肽，可以打破自我耐受并引起免疫反应。这一机制对控制人类和牲畜的传染病、开发抗癌疫苗和为自身免疫量身定做的疗法具有重要意义。目的、假设和实验方法我们假设，在非常规的细胞内转运过程中，新抗原可以被人类白细胞抗原-DP4获得。该项目的主要目标是表征这些新的抗原，并建立驱动它们在细胞表面显示的细胞生物学。为了支持这一目标，我们将使用支撑质谱学能力来定义天然MS对MHC的组合修饰，用定量MS绘制免疫蛋白质组图，并在两个系统(重组抗原提呈细胞和由案例合作伙伴开发的人源化小鼠黑色素瘤模型)中识别翻译后修饰的MHC多肽。学生还将接受广泛的实验室方法培训，包括共聚焦显微镜、组织培养、十二烷基硫酸钠-PAGE/蛋白质印迹、生化/细胞分析、分子生物学、生物信息学和统计计算(R)。学生将接受演讲技能和报告写作方面的培训，鼓励他们在国际会议上展示他们的工作，并将参加定期的期刊俱乐部。与BBSRC战略的相关性该项目既是好奇心主导的，又是创造性的，受到技术能力新发展的推动。该项目广泛地属于世界级支持生物科学的范围，并直接涉及一个系统方法，即通过重点推进定量和机械蛋白质组学技术，为生物科学和生物科学开发技术。