Mapping the antibody response to Trypanosoma brucei variant surface glycoprotein

绘制布氏锥虫变异表面糖蛋白的抗体反应

• 批准号: 10634694
• 负责人: Monica Mugnier
• 金额: $ 24.56万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-03 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10634694
• 关键词: Africa South of the Sahara; African Trypanosomiasis; Animal Diseases; Animals; Antibodies; Antibody Response; Antigenic Variation; Antigens; B-Lymphocytes; Bacteriophages; Binding; Blood; Cell surface; Chronic; Clone Cells; Collection; Data; Data Analyses; Detection; Diagnosis; Disease; Drug resistance; Economic Development; Epitope Mapping; Epitopes; Exposure to; Future; Genes; Genomics; High-Throughput Nucleotide Sequencing; Human; Immune; Immune Evasion; Immune response; Immune system; Immunity; Immunoprecipitation; Infection; Investigation; Libraries; Liquid substance; Maps; Measures; Membrane Glycoproteins; Methods; Molecular Conformation; Monoclonal Antibodies; Mus; Parasitemia; Parasites; Patients; Peptides; Phage Display; Phage ImmunoPrecipitation Sequencing; Play; Population; Positioning Attribute; Prevention; Process; Proteins; Proteome; Public Health; Resolution; Role; Sampling; Serum; Shapes; Specificity; Study models; Time; Tissues; Trypanosoma; Trypanosoma brucei brucei; Trypanosomiasis; Vaccines; Variant; Visit; Work; chronic infection; disorder prevention; extracellular; high throughput technology; immunogenic; insight; interest; mouse model; neglect; neglected tropical diseases; pathogen; response; tool

PROJECT SUMMARY Trypanosoma brucei is a causative agent of human and animal African trypanosomiasis, devastating diseases that endanger public health and present a major barrier to economic development in sub-Saharan Africa. The extracellular parasite manages to sustain an infection in the blood and tissues of its mammalian host by periodically “switching” its dense variant surface glycoprotein (VSG) coat. Drawing from a genomic repertoire of ~2000 VSG-encoding genes, the parasite changes its expressed VSG coat throughout infection. Before the host antibody response can clear a population expressing one VSG, the parasite switches expression to a new VSG, rendering it invisible to host antibody for a time. This process occurs continually and allows the parasite to maintain a chronic infection. The interaction between antibody and VSG, therefore, represents a critical interface in this infection. Despite its importance, the principles governing antibody recognition of VSG remain poorly understood. Here we propose to investigate the antibody-VSG interface during infection in high resolution using a high-throughput epitope mapping approach called phage immunoprecipitation sequencing (PhIP-seq). Using a phage display library containing peptides from every VSG ever annotated, we will measure antibody responses to VSG during chronic infections, while simultaneously using a high-throughput sequencing method developed by our lab to track the VSGs expressed during infection. In the first aim, we will characterize the dynamics and targets of anti-VSG antibody during chronic experimental infections. In the second aim, we will use PhIP-seq to map VSG epitopes in natural human infections. The proposed study will further our understanding of anti-VSG antibody responses while establishing T. brucei as a tractable model for the study of antibody responses to antigenically variable pathogens. Long term, this work could inform strategies for diagnosis or prevention of an important neglected tropical disease.

项目摘要 布鲁氏锥虫是人类和非洲锥虫病，毁灭性疾病的病因 这种危害了公共卫生，并构成了撒哈拉以南非洲经济发展的主要障碍。这 细胞外寄生虫通过 定期“切换”其致密的变体表面糖蛋白（VSG）涂层。从基因组曲目中汲取 在〜2000 VSG编码的基因中，寄生虫在整个感染过程中都会改变其表达的VSG外套。之前 宿主抗体反应可以清除表达一个VSG的种群，寄生虫将表达式切换到新的 VSG，使其无形地托管抗体一段时间。这个过程连续发生并允许寄生虫 保持慢性感染。因此，抗体与VSG之间的相互作用代表了关键 这种感染中的接口。尽管它的重要性，但管理VSG抗体识别的原则仍然存在 理解不佳。在这里，我们建议在高中感染期间研究抗体-VSG界面 使用称为噬菌体免疫沉淀测序的高通量表位映射方法的分辨率 （phip-seq）。使用噬菌体显示库，该库包含来自有史以来每个VSG的肽，我们将测量 慢性感染期间对VSG的抗体反应，同时使用高通量 我们实验室开发的测序方法是为了跟踪感染过程中表达的VSG。在第一个目标中，我们将 表征慢性实验感染过程中抗VSG抗体的动力学和靶标。在 第二个目的，我们将使用phip-seq绘制自然人类感染中的VSG表位。拟议的研究将 进一步，我们对抗VSG抗体反应的理解，同时建立t. brucei作为一种可探讨的模型 对抗原可变病原体的抗体反应的研究。长期，这项工作可以告知 诊断或预防重要被忽视的热带疾病的策略。