Project 2: B Cells

项目2：B细胞

基本信息

批准号：
10222106
负责人：
Scott Dexter Boyd
金额：
$ 93.92万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-23 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10222106
关键词：
2019-nCoV Acute Affinity Age Antibodies Antibody Response Antibody Specificity Antigens B-Cell Activation B-Lymphocyte Subsets B-Lymphocytes Bar Codes Biological Assay Biopsy Biopsy Specimen Blood Blood specimen COVID-19 Cell Differentiation process Cells Clinical Clinical Data Clinical Virology Clonal Evolution Clonal Expansion Clone Cells Communities DNA Data Data Analyses Databases Disease Effectiveness Epitopes Ethnic Origin Frequencies Funding Immune response Immunity Immunofluorescence Microscopy Immunoglobulin Genes Immunoglobulin Somatic Hypermutation Immunoglobulin-Secreting Cells Immunologic Receptors Individual Infection Joints Libraries Light Link Measures Memory B-Lymphocyte Monoclonal Antibodies Mucous Membrane Mutate Nose Patients Phenotype Plasma Cells Plasmablast Population Population Group Receptors, Antigen, B-Cell Resources Sampling Serologic tests Serological Serum Severity of illness Site Sorting - Cell Movement Specificity Specimen Structure Structure of mucous membrane of nose T cell response T-Lymphocyte TFRC gene Time Vaccination Vaccines Viral Antigens Virus Work Yeasts base cohort deep sequencing design expectation follow-up immune checkpoint blockade improved interest long term memory monoclonal antibody production neutralizing antibody patient response peptide based vaccine peripheral blood prognostic value response sex synergism transcriptome vaccine response

项目摘要

PROJECT 2: SUMMARY The B cell and plasma cell populations that give rise to serum and mucosal antibodies will ultimately determine the effectiveness and duration of an individual’s humoral immune response to SARS-CoV-2 infection. We will use several mutually-supporting strategies to analyze these cells in the Boyd lab: single B cell phenotyping, B cell receptor (BCR) deep sequencing and determination of antigen specificity with DNA-barcoded antigen tetramers; bulk B cell immunoglobulin gene repertoire sequencing; and monoclonal antibody production from antigen-specific B cells. In complementary strategy, the Jardetzky lab will make use of yeast display libraries of patient-derived single-chain antibody variable fragments (ScFv) enriched for native heavy-light chain pairing to determine the antigen specificity of hundreds to thousands of antigen-specific clones per patient. In longitudinal peripheral blood samples and nasal biopsy samples we will thoroughly characterize antigen-specific B cell clones in patient responses to SARS-CoV-2. We hypothesize that with these data we will be able to determine which features of B cell clonal responses to SARS-CoV-2 are associated with differences in COVID-19 disease severity and differences between populations groups stratified by age, sex, ethnicity and pre-existing conditions, or dysregulated immunity in the context of checkpoint blockade treatment. We further hypothesize that analysis of memory B cell populations together with serological responses may predict which individuals will have longer-lasting humoral protection against reexposure to SARS-CoV-2. Finally, we will evaluate the B cell responses stimulated by natural infection compared to vaccination, beginning with the Covaxx peptide- based vaccine cohort, but with the expectation that additional vaccines will be approved for use during the period of this project funding. In addition to studying aspects of the B cell responses that differ among these clinical scenarios, we will search for features such as “convergent” virus-specific BCRs of highly similar sequences shared between different individuals that may have prognostic value, for example by revealing that an individual has a potent neutralizing antibody response to SARS-CoV-2. Our Aims are the following: Specific Aim 1: Analyze B cell responses in acute COVID-19 disease. Specific Aim 2: Evaluate the formation of B cell memory to SARS-CoV-2. Specific Aim 3: Analyze mucosal B cell and plasma cell responses to SARS-CoV-2 compared to responses of B cells in the blood.

项目 2：总结产生血清和粘膜抗体的 B 细胞和浆细胞群将最终决定个体对 SARS-CoV-2 感染的体液免疫反应的有效性和持续时间。在 Boyd 实验室中使用几种相互支持的策略来分析这些细胞：单 B 细胞表型分析、B 细胞受体 (BCR) 深度测序和使用 DNA 条形码抗原确定抗原特异性四聚体；批量 B 细胞免疫球蛋白基因库测序和单克隆抗体生产；在补充策略中，Jardetzky 实验室将利用酵母展示库。患者来源的单链抗体可变片段 (ScFv) 富集天然重轻链配对纵向确定每个患者数百至数千个抗原特异性克隆的抗原特异性。外周血样本和鼻活检样本我们将彻底表征抗原特异性 B 细胞我们发现，通过这些数据，我们将能够确定患者对 SARS-CoV-2 的反应。 B 细胞对 SARS-CoV-2 克隆反应的哪些特征与 COVID-19 疾病的差异相关按年龄、性别、种族和既往史分层的人口群体之间的严重程度和差异条件，或在检查点封锁治疗背景下免疫失调。对记忆 B 细胞群和血清学反应的分析可以预测哪些个体将具有更持久的体液保护，防止再次暴露于 SARS-CoV-2 最后，我们将评估 B. 与疫苗接种相比，自然感染刺激的细胞反应，从 Covaxx 肽开始基于疫苗队列，但预计在此期间将批准使用更多疫苗除了研究 B 细胞反应的不同方面外，该项目资助的期间。临床场景中，我们将寻找高度相似的“趋同”病毒特异性 BCR 等特征不同个体之间可能具有预后价值的共享序列，例如通过揭示个体对 SARS-CoV-2 具有有效的中和抗体反应，我们的目标如下：具体目标 1：分析急性 COVID-19 疾病中的 B 细胞反应。具体目标 2：评估 B 细胞对 SARS-CoV-2 记忆的形成。具体目标 3：分析粘膜 B 细胞和浆细胞对 SARS-CoV-2 的反应与血液中的 B 细胞。