Molecular and Cellular Action of HLA Class II Molecules, Gen Risk Fctrs for VL

项目4：“人类白细胞抗原II类分子的分子和细胞作用，VL的主要遗传风险因素” 研究将以在当前TMRC期间所观察到的观察为基础，与第二阶段合作 Wellcome Trust Case Control Consortium(WTCCC2)，HLADR_DQ 11类区域包含主要 印度(和巴西)内脏利什曼病的遗传危险因素。 提出了四个具体目标： 1.测定活动期和既往患者全血和/或脾抽吸物中的转录信号 有风险的病例，以及Quantiferon和/或抗体阳性的无症状感染捐赠者(项目1) 与保护性HLADRB1等位基因组比较。 2.在体外和在体外刺激/感染细胞后，在mRNA和蛋白质水平上确定， 顺式调节多态是否是风险和保护性的HLADRB1等位基因的功能性关联 组。 3.从EBV中提纯天然加工的利什曼多肽，并用质谱仪进行鉴定 HLADRB1*13/*14制备的转化B细胞与全反式刺激的HLADRB1*15供者的比较 从当地的印度菌株多诺瓦尼乳杆菌中制备的粗制寄生虫裂解物，与电子预测中的 表位和结合亲和力，并开发一种通过接种疫苗来颠覆免疫反应的策略 4.通过检测抗原提呈细胞功能和免疫应答来验证AIM 3的结果 暴露人群中自然处理和预测的多肽表位，包括现症和既往病例， 和无症状的受感染捐赠者。 相关性(请参阅说明)： 项目4.这些研究将有助于理解人类白细胞抗原分类的分子和细胞基础 基因是疾病的主要遗传风险因素，并对治疗学和疫苗有影响 设计和交付。

Project 4: "Molecular and cellular action of HLA class II molecules, the major genetic risk factors for VL" The studies will build on observations made during the current TMRC, in collaboration with Phase 2 of the Wellcome Trust Case Control Consortium (WTCCC2), that HLA DR_DQ class 11 region contains the major genetic risk factor for visceral leishmaniasis in India (and Brazil). Four specific aims are proposed: 1. To determine transcriptional signatures in whole blood and/or splenic aspirates from active and past cases, and Quantiferon and/or antibody positive asymptomatic infected donors (Project 1), carrying risk versus protective HLA DRB1 allele groups. 2. To determine at mRNA and protein levels ex vivo, and following stimulation/infection of cells in vitro, whether cis-acting regulatory polymorphisms are functional correlates of risk and protective HLA DRB1 allele groups. 3. To purify, and identify using mass spectrometry, naturally processed leishmanial peptides from EBV transformed B cells prepared from HLA DRB1*13/*14 versus HLA DRB1*15 donors stimulated with whole crude parasite lysate prepared from a local Indian strain of L. donovani, compare with in silico predictions of epitopes and binding affinity, and develop a strategy to subvert the immune response by vaccination 4. To validate results of aim 3 by measuring antigen presenting cell function and immune response to naturally processed and predicted peptide epitopes in exposed individuals, including active and past cases, and asymptomatic infected donors. RELEVANCE (See instructions): Project 4. These studies will contribute to understanding of the molecular and cellular basis of HLA class genes as the major genetic risk factors for disease, and have implications for therapeutics and vaccine design and delivery.