Defining serologic correlates of human hookworm infection

项目总结 钩虫感染是贫穷国家贫血、营养不良和生长迟缓的主要原因，尤其是 在撒哈拉以南非洲，那里有数百万人被感染。目前控制钩虫的策略依赖于 大规模药监局使用驱虫药，尽管证据令人质疑其长期性 这一方法在可持续控制或消除高危人群中钩虫方面的有效性。自2007年以来， 耶鲁大学和加纳大学野口纪念医学研究所 合作确定钩虫在流行社区的流行病学特征。这些研究已经 确定与钩虫感染状况和驱虫治疗反应相关的因素。初步 使用加纳北部金坦波人类样本的数据表明，宿主抗体(IgG)水平直接针对 钩虫成虫排泄/分泌(ES)蛋白与活动性感染密切相关。然而， 到目前为止，对钩虫抗原的这些抗体反应的靶点知之甚少，钩虫抗原是一种主要的 对宿主-寄生虫相互作用和发病机制的了解存在差距。我们假设宿主抗体是针对 特定的钩虫蛋白可以预测感染状态，并且一类特定的蛋白，即 过敏原是这种反应的驱动力。针对特定目标1的研究将验证免疫球蛋白之间的相关性 利用收集的1,002份人血清样本和人口学数据分析抗体水平和感染状况 2007-2020年间在加纳进行实地考察。抗体水平将通过酶联免疫吸附试验进行测量和分析 粪便显微镜检查与钩虫感染状态的关系。来自人体研究的数据将 与使用Necator的仓鼠模型进行的控制感染和治疗研究的结果进行比较 美洲豹。AIM 2的研究将集中于识别宿主抗体反应的特定蛋白质靶标 利用重组蛋白表达和无标记定量蛋白质组学。候选钩虫变应原 将表达和纯化蛋白质，然后使用人和 在目标1中描述的仓鼠血清样本。此外，美洲钩虫成虫ES蛋白将是 用高反应血清池进行免疫沉淀，然后进行LC-MS/MS分离和多肽图谱 与钩虫/蠕虫基因组序列数据库进行比较。这些实验将检验这一假设 钩虫变应原是与活动性感染相关的宿主免疫反应的抗原驱动因素。 这项研究的主要目标是(1)将抗体反应与感染状况联系起来。 以及(2)钩虫变应原和新型抗原蛋白在宿主免疫反应中的作用。 这些研究利用了加纳大学和耶鲁大学之间的长期、富有成效的合作 这导致了新的初步数据的产生和第一个非洲钩虫株在 实验室模型。这些研究将加深我们对钩虫发病机制的了解，并为我们提供 开发创新工具，以监测流行社区的驱虫计划。

PROJECT SUMMARY Hookworm infection is a leading cause of anemia, malnutrition and growth delay in poor countries, especially in sub-Saharan Africa, where millions of people are infected. Current strategies to control hookworm rely on Mass Drug Administration of anthelminthic drugs, although evidence calls into question the long-term effectiveness of this approach to sustainably control or eliminate hookworm in populations at risk. Since 2007, Yale University and the Noguchi Memorial Institute for Medical Research at the University of Ghana have collaborated to characterize the epidemiology of hookworm in endemic communities. These studies have identified factors associated with hookworm infection status and response to deworming treatment. Preliminary data using human samples from Kintampo North, Ghana, suggests that host antibody (IgG) levels directed at hookworm adult worm excretory/secretory (ES) proteins are closely correlated with active infection. However, to date little is known about the targets of these antibody responses to hookworm antigens, which is a major gap in understanding of host-parasite interactions and pathogenesis. We hypothesize that host antibodies to specific hookworm proteins are predictive of infection status, and that a particular class of proteins, namely allergens, are the drivers of this response. Studies in Specific Aim 1 will validate the correlation between IgG antibody levels and infection status using 1,002 human serum samples and demographic data collected between 2007-2020 in Ghana field studies. Antibody levels will be measured by ELISA and analyzed for associations with hookworm infection status as determined by fecal microscopy. Data from human studies will be compared to results from controlled infection and treatment studies using the hamster model of Necator americanus. Studies in Aim 2 will focus on identifying the specific protein targets of host antibody responses using recombinant protein expression and Label Free Quantitative proteomics. Candidate hookworm allergen proteins will be expressed and purified, followed by screening for immunoreactivity using the human and hamster serum samples described in Aim 1. In addition, N. americanus adult worm ES proteins will be immunoprecipitated using highly reactive serum pools, followed by LC-MS/MS isolation and peptide mapping against hookworm/helminth genome sequence databases. These experiments will test the hypothesis that hookworm allergens are antigenic drivers of the host immune response that is associated with active infection. The overarching goals of the proposed research are to (1) correlate antibody responses with infection status and (2) characterize the role of hookworm allergens and novel antigenic proteins in host immune responses. These studies leverage a longstanding, productive collaboration between the University of Ghana and Yale that led to generation of novel preliminary data and adaptation of the first African hookworm strain in a laboratory model. These studies will enhance our understanding of hookworm pathogenesis and inform development of innovative tools to monitor deworming programs in endemic communities.