Simultaneous antigen receptor repertoire profiling and single-cell transcriptomics in T and B lymphocytes from limited clinical samples

对有限临床样本中的 T 和 B 淋巴细胞进行同步抗原受体库分析和单细胞转录组学分析

• 批准号: 8971431
• 负责人: Steven Edward Bosinger
• 金额: $ 39.96万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-26 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8971431
• 关键词: Antibodies; Antigen Receptors; Antigens; Antiviral Response; Area; B-Lymphocytes; Base Sequence; Benchmarking; Binding; Biological; Biological Assay; Biopsy Specimen; Blood; Bone Marrow; CD8B1 gene; Cells; Clinical; Clinical Trials; Cloning; Collection; Computational algorithm; Data; Development; Failure; Gene Expression Profile; Genes; Genomics; Goals; Human; Immune response; Immune system; Immunoglobulins; Individual; Infection; Influenza vaccination; Light; Link; Measures; Messenger RNA; Methodology; Methods; Monitor; Population; Procedures; Protocols documentation; RNA Sequences; Reading; Retrieval; Sampling; Sequence Analysis; System; T cell response; T-Lymphocyte; Technical Expertise; Techniques; Testing; Tissue-Specific Gene Expression; Translating; Vaccination; Vaccines; Validation; Work; arm; base; clinically relevant; cost; cost effectiveness; high throughput screening; high throughput technology; improved; insight; next generation sequencing; prospective; public health relevance; receptor; rectal; single cell sequencing; success; transcriptome sequencing; transcriptomics; vaccine development; vaccine trial; vaccinology

 DESCRIPTION (provided by applicant): In the vast majority of vaccine trials, the primary readouts are empirical do not collect information about the immunological determinants of a vaccine's success or failure. In recent years, several vaccine studies have combined high-throughput transcriptomic data with measures of antigenicity. These "systems vaccinology" studies have proven invaluable insight into the determinants of efficacy and antigenicity, and have demonstrated the enormous value in mechanistic studies of human vaccination. However, one area that is particularly understudied is the composition of antigen specific receptors that arise during successful and unsuccessful vaccinations. Collection of antigen-specific clonotype information in clinical vaccine studies would provide valuable data that could be used to accelerate vaccine development. The utility of current methodology for repertoire sequencing for immunological studies has been limited due to several factors (i) high-cost and low-throughput of cloning based assays (ii) most high-throughput assays only provide information on a single gene(typically the H-chain of immunoglobulin or ß-chain for TCRs), or (iii) do not link ag-receptor sequences with immunophenotypic or transcriptomic data. In preliminary work, we have developed a protocol that simultaneously queries the transcriptome and paired antigen receptor sequences in B lymphocytes derived from human bone marrow after flu vaccination using next generation sequencing. The goal of this proposal is to complete development of this combined "-Seq" assay for both B and T cells, including establishment of its limitations and benchmark against contemporary repertoire sequencing techniques.

 描述（由适用提供）：在绝大多数疫苗试验中，主要读数是经验，请勿收集有关疫苗成功或失败的免疫确定剂的信息。近年来，一些疫苗研究将高通量转录组数据与抗原性测量结合在一起。这些“系统疫苗”研究证明了对效率和抗原性决定者的宝贵见解，并证明了人类疫苗的机械研究中具有巨大的价值。但是，尤其了解的一个区域是在成功和失败的疫苗过程中的抗原特异性受体的组成。临床疫苗研究中抗原特异性饼干类型信息的收集将提供可用于加速疫苗开发的有价值数据。由于几个因素（i）基于克隆的测定的高成本和低通量，目前的方法对免疫研究的曲目测序的实用性受到限制，大多数高通量测定只提供单个基因的信息（通常是单个基因的信息）（通常是TCRS的H-chain fr Imn.imii imecipt，或IIII III，或III II I.-RINKITER，或者III III not（III III）。转录组数据。在初步工作中，我们开发了一项方案，该方案仅查询转录组和配对的抗原受体序列，使用下一代测序在流感后源自人骨髓的B淋巴细胞中。该提案的目的是完成B和T细胞的这种组合“ -Seq”测定的开发，包括建立其局限性和针对当代曲目测序技术的基准。