PROCESSED ANTIGEN CHARACTERIZATION BY MASS SPECTROMETRY

疾病相关抗原的鉴定通常是开发疫苗或疫苗制剂的重要步骤。 有效对抗细菌和病毒感染、癌症、自身免疫 疾病，移植排斥和生物恐怖主义制剂。这里提出的研究是为了继续 开发用于呈递给免疫原的肽抗原的序列分析的仪器和方法， 与主要组织相容性复合体的I类和II类分子相关的免疫系统。 在阿托摩尔水平上存在的疾病相关抗原的常规检测和表征， 10，000个自身肽的混合物是目标。在拟议的研究中，我们将实施我们最近的 开发了峰值停车技术和纳米流高性能液相色谱技术， 仪器、线性阱质谱仪和串联线性阱-傅里叶变换质谱仪 光谱仪我们还将继续开发差分显示软件包， 可以比较由健康和患病细胞呈递的I类和II类肽，然后 对这些肽进行测序，这些肽是疾病的独特特征，并且可能被识别为 免疫系统的抗原。拟议研究的其他目的包括：（1） 鉴定I型糖尿病中致病性CD 8（+）T细胞识别的其他I类肽，（2） 鉴定与移植物抗宿主病和组织相关的其它次要组织相容性抗原 移植排斥，（3）以确定磷酸化的I类肽的表面上唯一提出的 癌细胞，（4）鉴定由异戊二烯化的乳腺癌抗原呈递的抗原性I类肽， C35，（5）鉴定肾细胞癌上受I类HLA分子限制的肽抗原，（6） 鉴定黑色素瘤细胞上呈递的HLA A3限制性和共有抗原，和（7）评估 酪氨酸酶的I类和II类表位的加工和呈递机制， 黑色素瘤中T细胞反应的靶点。

Identification of disease associated antigens is an often an important step in the development of vaccines or other immune system modulators that are effective against bacterial and viral infections, cancer, autoimmune disorders, transplant rejection and bioterrorism agents. Proposed here is research to continue the development of instrumentation and methods for the sequence analysis of peptide antigens presented to the immune system in association with class I and class II molecules of the major histocompatibility complex. Routine detection and characterization of disease associated antigens present at the attomole level in a mixture of 10,000 self peptides is the goal. In the proposed research, we will implement our recently developed peak parking technology and nano-flow high performance liquid chrornatography to two new instruments, the linear trap mass spectrometer and the tandem linear trap-Fourier transform mass spectrometer. We will also continue to development the differential display software package which makes it possible to compare class I and class II peptides presented by healthy and diseased cells and then to sequence those peptides that are uniquely characteristic of the disease and likely to be recognized as antigens by the immune system. Additional aims of the proposed research include the following: (1) to identify additional class I peptides recognized by pathogenic CD8(+) T-cells in Type I diabetes, (2) to identify additional minor histocompatibility antigens associated with graft-vs-host disease and tissue transplant rejection, (3) to identify phosphorylated class I peptides presented uniquely on the surface of cancer cells, (4) to identify antigenic class I peptides presented from the prenylated breast cancer antigen, C35, (5) to identify peptide antigens restricted by class I HLA molecules on renal cell carcinoma, (6) to identify an HLA A3 restricted and shared antigen presented on melanoma cells, and (7) to evaluate the processing and mechnanism of presentation for both class I and class II epitopes from tyrosinase that are targets of T cell responses in melanoma.