PROCESSED ANTIGEN CHARACTERIZATION BY MASS SPECTROMETRY

通过质谱法进行抗原表征

• 批准号: 6373317
• 负责人: DONALD F HUNT
• 金额: $ 53.85万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6373317
• 关键词: HY antigen; MHC class I antigen; MHC class II antigen; antigen presentation; bioimaging /biomedical imaging; biomedical equipment development; capillary electrophoresis; cross immunity; electrospray ionization mass spectrometry; graft versus host disease; high performance liquid chromatography; histocompatibility antigens; melanoma; multiple sclerosis; myelin basic proteins; protein sequence; tumor antigens

DESCRIPTION: (adapted from applicant's abstract) 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 is proposed. Routine detection and characterization of disease associated antigens present at the attomole level in a mixture of 10,000 self peptides is the expected outcome. Identification of disease associated antigens is an important first step in the development of vaccines or other immune system modulators that are effective against bacterial and viral infections, cancer, autoimmune disorders and tissue transplant rejection. Multistage chromatography in conjunction with our recently developed, automated peak parking technology plus nanoliter/min, high performance liquid chromatography and high performance capillary electrophoresis interfaced to an ion trap mass spectrometer via an electrospray ionization source will be employed to achieve this goal. Specific aims of the proposed research include the following: (1) to identify melanoma specific antigens presented by the class I molecules, HLA A2.1, A1 and A3, and the class II molecule, DR4, (2) to identify minor histocompatibility antigens associated with graft-vs-host disease and tissue transplant rejection, (3) to identify peptides derived from myelin basic protein as well as cross reactive self peptides that are presented by the multiple sclerosis associated class II molecule, DR15Dw2, and recognized by T-cell clones derived from multiple sclerosis patients, (4) to identify peptides presented by the class I molecule, B27, that are unique to individuals diagnosed with ankylosing spondylitis (5) to identify peptides presented by the class I molecule, A2.1, that are unique to individuals diagnosed as having chronic lymphocytic leukemia, (6) to identify and characterize peptides presented by HLA-A2.1, that have been post translationally modified by either phosphorylation or glycosylation, (7) to identify class II peptides presented by the HLA-DR4Dw4 molecule in the presence and absence of the chaperone molecule, HLA-DM.

描述：（改编自申请人的摘要）继续研究 肽序列分析仪器和方法的发展 呈递给免疫系统的与I类和II类相关的抗原 II分子的主要组织相容性复合体的建议。常规 检测和表征存在于细胞中的疾病相关抗原 10，000个自身肽的混合物中的阿托摩尔水平是预期的结果。 疾病相关抗原的鉴定是免疫治疗中重要的第一步。 开发有效的疫苗或其他免疫系统调节剂 针对细菌和病毒感染、癌症、自身免疫性疾病和组织 移植排斥反应多级色谱结合我们的 最近开发的自动高峰停车技术加上纳升/分钟，高 高效液相色谱和高效毛细管 通过电喷雾与离子阱质谱仪连接的电泳 将使用电离源来实现该目标。的具体目标 建议的研究包括以下内容：（1）确定黑色素瘤特异性 由I类分子HLAA2.1、A1和A3呈递的抗原，以及 II分子，DR 4，（2）以鉴定相关的次要组织相容性抗原 移植物抗宿主病和组织移植排斥反应，（3）识别 来自髓鞘碱性蛋白的肽以及交叉反应性自身 由多发性硬化症相关II类 分子DR 15 Dw 2，并被来自多种细胞的T细胞克隆识别。 硬化症患者，（4）鉴定由I类分子呈递的肽， B27，这是唯一的个人诊断为强直性脊柱炎（5） 鉴定由I类分子A2.1呈递的肽， 诊断为患有慢性淋巴细胞白血病的个体，（6） 鉴定和表征由HLA-A2.1呈递肽， 通过磷酸化或糖基化修饰，（7） 鉴定HLA-DR 4Dw 4分子在存在下呈递的II类肽， 以及缺少分子伴侣HLA-DM。