IDENTIFICATION OF MODIFIED AND NATURAL HEL PEPTIDE FRAGMENTS PRESENTED BY MHC

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 T细胞的免疫应答需要识别抗原。 我们已经证明了之前与模型蛋白抗原鸡蛋溶菌酶（HEL），这种蛋白质的反应涉及解折叠和降解的肽被识别的抗原呈递细胞（APC）的表面上与主要组织相容性复合体（MHC）分子。 先前的结果表明，T细胞在被I-Ak型APC激活后，对合成的10聚体残基HEL 51-62产生应答。 类似地，T细胞在被I-Ek型APC激活后，对13聚体HEL 84-96产生应答。 用HEL处理后从APC的MHC II类复合物中提取的混合物的HPLC色谱得到免疫活性组分，其通过MALDI-TOF和cf-FAB MS分析。I-Ak型APC的主要组分为HEL 48-63、48-62、48-61，I-Ek型APC的主要组分为HEL 84-97、84-98、84-102。为了扩展对产生呈递给T细胞的肽的蛋白抗原加工机制的理解，我们正在研究从一系列突变的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。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.