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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 T细胞的免疫反应需要识别抗原。 我们以前已经使用模型蛋白抗原母鸡蛋白溶菌酶（HEL）证明了这种对蛋白质的反应涉及与主要的组织相容性复合物（MHC）分子相关的在抗原呈递细胞（APC）表面上识别的肽。 先前的结果表明，I-AK型APC激活后T细胞对合成的10-Mer残基响应HEL 51-62。 同样，I-EK型APC激活后的T细胞对13-MER的HEL 84-96响应。 用HEL处理后，从APC的MHC II类配合物中提取的混合物的HPLC色谱法得到了通过MALDI-TOF和CF-FAB MS分析的免疫活性馏分。 I-AK型APC和HEL 84-97、84-98、84-102的主要组件是HEL 48-63、48-62、48-61，用于I-EK型APC。为了扩展对产生呈现给T细胞的肽的蛋白质抗原加工机制的理解，我们正在努力表征由一系列突变的HEL蛋白产生的肽。 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。