Project 2: B Cells

项目2：B细胞

• 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
• 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

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：摘要 产生血清和粘膜抗体的B细胞和浆细胞群最终将确定 一个人对SARS-COV-2感染的体液免疫反应的有效性和持续时间。我们将 使用几种相互支持的策略在博伊德实验室中分析这些细胞：单个B细胞表型，b 细胞受体（BCR）深层测序和用DNA-barcoded抗原的抗原特异性测定 四聚体；散装B细胞免疫球蛋白基因曲目测序；和单克隆抗体产生 抗原特异性B细胞。在完整的策略中，Jardetzky实验室将利用酵母显示库 患者衍生的单链抗体可变片段（SCFV）富含天然重链链配对 确定每位患者数百至数千个抗原特异性克隆的抗原特异性。在纵向中 外周血样品和鼻活检样品我们将彻底表征抗原特异性B细胞 患者对SARS-COV-2的反应中的克隆。我们假设有了这些数据，我们将能够确定 B细胞克隆反应对SARS-COV-2的哪些特征与Covid-19疾病的差异有关 人口群体之间的严重性和差异，按年龄，性别，种族和现有 在检查点阻滞治疗的情况下，情况或免疫学失调。我们进一步假设 对记忆B细胞群体的分析以及血清学反应可以预测哪些人 将对重新暴露于SARS-COV-2的持续持久防护保护。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胡椒开始 基于疫苗队列，但期望在此期间批准使用其他疫苗 该项目资金的期限。除了研究B细胞反应的各个方面之外 临床场景，我们将搜索高度相似的“收敛性”病毒特异性BCR 在不同的个体之间共享可能具有原型值的序列，例如通过揭示 一个人对SARS-COV-2具有潜在的中和抗体反应。我们的目标是以下： 具体目标1：分析急性共卷19疾病中的B细胞反应。 特定目标2：评估B细胞记忆对SARS-COV-2的形成。 特定目标3：分析粘膜B细胞和浆细胞对SARS-COV-2的反应 血液中的B细胞。