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

项目4：“HLA II类分子的分子和细胞作用，VL的主要遗传风险因素” 研究将建立在当前TMRC期间所做的观察的基础上，与第二阶段合作， 欢迎Trust Case Control Consortium（WTCCC 2），HLA DR_DQ 11类区域包含主要的 在印度（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.为了确定来自活动性和过去的患者的全血和/或脾抽吸物中的转录特征， 病例和Quantiferon和/或抗体阳性无症状感染供体（项目1），存在风险 与保护性HLA-DRB 1等位基因组相比。 2.为了在离体mRNA和蛋白质水平以及在体外细胞刺激/感染后测定， 顺式作用调节多态性是否是风险和保护性HLA-DRB 1等位基因的功能相关性 组 3.纯化并使用质谱法鉴定来自EBV的天然加工的利什曼肽 从HLA DRB 1 *13/*14与HLA DRB 1 *15供体制备的转化的B细胞用全 从当地的印度L. donovani，与计算机预测的 表位和结合亲和力，并制定一项战略，以颠覆免疫反应的疫苗接种 4.通过检测抗原呈递细胞功能和对 暴露个体中天然加工和预测的肽表位，包括活动和既往病例， 和无症状感染者。

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 Welcome 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.