IDENTIFICATION OF MODIFIED AND NATURAL HEL PEPTIDE FRAGMENTS PRESENTED BY MHC

MHC 呈现的修饰和天然 HEL 肽片段的鉴定

• 批准号: 8361330
• 负责人: EMIL Raphael UNANUE
• 金额: $ 2.68万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361330
• 关键词: Aftercare; Antigen-Presenting Cells; Antigens; Chromatography; Complex; Funding; Grant; Hen Egg Lysozyme; High Pressure Liquid Chromatography; Histocompatibility Antigens Class II; Immunology; Major Histocompatibility Complex; Mass Spectrum Analysis; Methods; Modeling; Mutate; National Center for Research Resources; Peptide Fragments; Peptides; Principal Investigator; Proteins; Research; Research Infrastructure; Resources; Series; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Surface Antigens; T cell response; T-Lymphocyte; United States National Institutes of Health; Work; antigen processing; biomedical resource; cost; nano-electrospray; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The immuno response of T cells requires an antigen to be recognized. We have demonstrated before with the model protein antigen Hen Egg Lysozyme (HEL), that this response to proteins involves unfolding and degradation to peptides that are recognized on the surface of antigen-presenting cells (APC) in association with the major histocompatibility complex (MHC) molecules. Previous results showed that T-cells, after activation by I-Ak type APC, respond to a synthetic 10-mer residue, HEL 51-62. Similarly, T cells after activation by I-Ek type APC, respond to a 13-mer, HEL 84-96. HPLC chromatography of the mixture extracted from MHC class II complexes of APC after treatment with HEL gave the immuno active fractions that were analyzed by MALDI-TOF and cf-FAB MS. The major components were HEL 48-63, 48-62, 48-61 for I-Ak type APC and HEL 84-97, 84-98, 84-102 for I-Ek type APC. To extend the understanding of the protein antigen processing mechanism producing the peptides that are presented to the T cells, we are working on characterizing peptides produced from a series of mutated HEL proteins. We are developing methods involving MALDI, and electrospray and nano-electrospray tandem MS to solve these important immunology problems.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 T 细胞的免疫反应需要抗原才能被识别。 我们之前已经用模型蛋白抗原鸡蛋溶菌酶 (HEL) 证明，这种对蛋白质的反应涉及到肽的解折叠和降解，这些肽在抗原呈递细胞 (APC) 表面上与主要组织相容性复合体 (MHC) 分子相关识别。 先前的结果表明，T 细胞在被 I-Ak 型 APC 激活后，会对合成的 10 聚体残基 HEL 51-62 做出反应。 同样，T 细胞在被 I-Ek 型 APC 激活后，会对 13 聚体 HEL 84-96 做出反应。 用 HEL 处理后，对从 APC 的 MHC II 类复合物中提取的混合物进行 HPLC 色谱分析，得到免疫活性级分，并通过 MALDI-TOF 和 cf-FAB MS 进行分析。 I-Ak 型 APC 的主要部件为 HEL 48-63、48-62、48-61，I-Ek 型 APC 的主要部件为 HEL 84-97、84-98、84-102。为了加深对产生呈递给 T 细胞的肽的蛋白质抗原加工机制的理解，我们正在研究由一系列突变的 HEL 蛋白产生的肽的特征。 我们正在开发涉及 MALDI、电喷雾和纳米电喷雾串联 MS 的方法来解决这些重要的免疫学问题。