Development and maintenance of human glycan and phospholipid antibody repertoires

人聚糖和磷脂抗体库的开发和维护

基本信息

批准号：
10396001
负责人：
John Franklin Kearney
金额：
$ 44.45万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10396001
关键词：
Affinity Allergic Disease Antibodies Antibody Repertoire Antibody Specificity Antigenic Specificity Antigens Apoptosis Atypical lymphocyte Autoantigens Autoimmune Autologous Automobile Driving Avidity B cell repertoire B cell therapy B-Cell Antigen Receptor B-Cell Development B-Lymphocyte Subsets B-Lymphocytes Bacteria Binding Biochemical Biological Blood Cadaver Carbohydrates Cell Compartmentation Clone Cells Cloning Development Drug or chemical Tissue Distribution Epitopes Evolution Excision Exhibits Gene Expression Gene Library Gene Rearrangement Genetic Goals Heterophile Antigens Homeostasis Host Defense Housekeeping Human Immune Immune response Immune system Immunity Immunization Immunoglobulin Class Switching Immunoglobulin Gene Rearrangement Immunoglobulin Genes Immunoglobulin Isotypes Immunoglobulin Somatic Hypermutation Immunoglobulins Immunotherapy Incidence Individual Infection Libraries Maintenance Mediating Memory Memory B-Lymphocyte Molecular Mucous Membrane Mus Natural regeneration Organ Organism Outcome Pathogenicity Patients Phenotype Phospholipids Polysaccharides Property Recombinants Recovery Role Sampling Serum Shapes Site Somatic Mutation Sorting - Cell Movement Specificity Surface T-Lymphocyte Testing Tissue Donors Tissue Transplantation Tissues Vaccine Therapy Xenograft procedure allograft rejection anti-CD20 base chronic infection clinically relevant cohort design experience host colonization host microbiota human tissue immune function indexing microbiota microorganism antigen mouse model natural antibodies neoantigens novel novel therapeutics pathogen prevent programs recurrent infection rituximab selective IgM deficiency senescence transcriptome

项目摘要

Project 1: Development and maintenance of human glycan and phospholipid antibody repertoires Project Summary Natural antibodies (nAbs) exist in the blood of multiple mammalian species in the absence of deliberate immunization. Reactivity of nAbs with epitopes conserved between pathogens and autologous host antigens allow these antibodies to perform dual functions in immunity: providing important host defense against infection and facilitating housekeeping functions important for tissue homeostasis. The mechanisms controlling human nAb development and maintenance, however, are poorly understood. The goal of this project is to define mechanisms controlling maturation of the human natural B lymphocyte repertoire and its tissue distribution. We will achieve this goal by completing a targeted analysis of B cells reactive with conserved carbohydrate and phospholipid T lymphocyte-independent antigens associated with clinically relevant bacteria and xenoantigens. Through this approach, we will test our central hypothesis that the selection of innate-like B cell clonotypes and antigen-specific tuning of the nAb-producing B cell repertoire depend on interactions with autologous antigens and microbial antigens encountered at mucosal surfaces, which together modulate the entry of B cell clonotypes into the memory and antibody-secreting B cell compartments. In the first Specific Aim, we will sort-purify single, indexed carbohydrate- and phospholipid-binding B cells from a cohort of cadaveric human tissue donors. Immunoglobulin gene expression in these B cells will be analyzed together with expressed cellular phenotype to determine the distribution of clonal networks of nAb-producing B cells across B cell compartments in multiple human tissues. We will additionally examine the stability of innate-like B cell clonotypes and specificity of nAb repertoire by longitudinally sampling human blood after anti-CD20 (rituximab) B cell depletion, to determine the extent of antigen-reactive clonal B cell extirpation and clonal repertoire recovery during B cell compartment regeneration. Specific Aim 2 will utilize a novel, high-throughput antibody-cloning and expression platform to express immunoglobulin gene rearrangements from gene amplicon libraries as recombinant Abs and examine their binding properties, including antigen affinity and fine specificity. We will additionally examine the effects of somatic mutation on the binding properties of nAb by assessing the global reactivity of cloned antibodies and germline-reverted clonotypes using mammalian glycan antigen microarrays. This targeted analysis of antigen- reactive human B cells will permit analysis of fine antigen-specificity, affinity, and avidity of B cell clonotypes across human tissues, and the determination of whether these features of the BCR influence the tissue, B cell subset, and immunoglobulin isotype distribution of certain clones. Because nAb antigens are expressed by multiple commensal and pathogenic organisms and are protective in normal immune homeostasis, such findings will facilitate the development of immunotherapies designed to intersect the natural repertoire.

项目1：人类血糖和磷脂抗体库的开发和维护项目摘要在没有故意的情况下，在多种哺乳动物物种的血液中存在天然抗体（NABS）免疫。 NAB的反应性具有病原体和自体宿主抗原之间保守的表位的反应性允许这些抗体在免疫中执行双重功能：为感染提供重要的宿主防御并促进对组织稳态重要的家政功能。控制人类的机制但是，NAB的开发和维护知之甚少。该项目的目的是定义控制人天然B淋巴细胞库及其组织分布的成熟的机制。我们通过完成保守的碳水化合物反应性的B细胞的目标分析，将实现此目标与临床相关细菌和异种抗原相关的磷脂T淋巴细胞非依赖性抗原。通过这种方法，我们将测试我们的中心假设，即选择先天性B细胞clonotypes和产生NAB的B细胞库的抗原特异性调整取决于与自体抗原的相互作用和在粘膜表面遇到的微生物抗原，共同调节B细胞clonotypes的进入进入记忆和分泌B细胞室的抗体。在第一个特定目的中，我们将分类单一纯化，索引的碳水化合物和磷脂结合B细胞来自尸体人体组织供体的队列。这些B细胞中的免疫球蛋白基因表达将与表达的细胞表型一起分析确定多个B细胞室跨B细胞室中NAB产生B细胞的克隆网络的分布人体组织。我们还将研究先天性B细胞clonotypes的稳定性和NAB的特异性抗CD20（Rituximab）B细胞耗竭后，纵向采样人血的曲目，以确定 B细胞室期间的抗原反应性克隆B细胞消除和克隆曲目恢复的程度再生。特定的目标2将利用一种新颖的，高通量的抗体克隆和表达平台从基因扩增子库中表达免疫球蛋白基因重排作为重组ABS并检查它们的结合特性，包括抗原亲和力和良好的特异性。我们还将研究通过评估克隆抗体的整体反应性和使用哺乳动物聚糖抗原微阵列进行种系的clonotypes。抗原的针对性分析反应性的人B细胞将允许分析B细胞clonotypes的精细抗原特异性，亲和力和亲和力在整个人体组织中，确定BCR的这些特征是否影响组织B细胞是否影响子集和某些克隆的免疫球蛋白同种型分布。因为Nab抗原由多种共生和致病生物，在正常的免疫稳态中具有保护性，此类发现将促进旨在与自然曲目相交的免疫疗法的发展。