Effects of neonatal microbial exposure on anti-polysaccharide B cell development

新生儿微生物暴露对抗多糖 B 细胞发育的影响

• 批准号: 10218018
• 负责人: John Franklin Kearney
• 金额: $ 37.13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218018
• 关键词: 2 year old; Adenoidal structure; Adult; Age; Allergens; Allergic Disease; Antibodies; Antibody Repertoire; Antibody Response; Antibody Specificity; Antigenic Specificity; Antigens; Autoantigens; Autoimmune Diabetes; Autoimmune Diseases; B cell repertoire; B-Cell Antigen Receptor; B-Cell Development; B-Lymphocyte Epitopes; B-Lymphocytes; Bacteria; Bacterial Infections; Bacterial Polysaccharides; Binding; Biochemical; Blood; Bone Marrow; Cell Compartmentation; Cell Wall; Cell surface; Child; Chimera organism; Clonal Expansion; Clone Cells; Development; Dextrans; Environment; Environmental Risk Factor; Epitopes; Evolution; Exposure to; Frequencies; Funding; Germ-Free; Glucosamine; Gnotobiotic; Goals; Heavy-Chain Immunoglobulins; Heterogeneity; Human; Immune; Immunization; Immunoglobulin Genes; Immunoglobulin M; Individual; Infant; Infection; Inflammatory Response; Kinetics; Lamina Propria; Licensing; Life; Lymphoid; Lymphopoiesis; Mammals; Manuscripts; Mediating; Modeling; Mouse Strains; Mus; Nature; Neonatal; Organism; Outcome Study; Pathogenicity; Phenotype; Phospholipids; Phosphorylcholine; Polysaccharides; Predisposition; Preparation; Probiotics; Process; Property; Recombinant Antibody; Recombinants; Risk; Role; Serum; Shapes; Signal Transduction; Specificity; Streptococcus pneumoniae; Streptococcus pyogenes; System; T-Independent Antigens; Testing; Therapeutic Intervention; Time; Tonsil; Transgenic Mice; Transgenic Organisms; Umbilical Cord Blood; Vaccination; Vaccines; age group; allergic airway disease; autoreactive B cell; bacterial resistance; base; commensal microbes; disorder prevention; experience; gut colonization; gut microbiota; insight; microbial; microbial colonization; microbiota; microorganism antigen; natural antibodies; neonatal exposure; neonate; novel; novel therapeutics; pathogen; pathogenic bacteria; peripheral blood; receptor; reconstitution; response

ABSTRACT Innate-like B cell-derived natural antibodies (NAbs) against T-independent polysaccharide (PS) and phospholipid epitopes are universally present in mammals and serve important roles in the protection against many bacterial pathogens. However robust antibody responses against PS antigens following infection or immunization are absent in neonates. We previously showed that neonatal exposure to PS and phospholipid antigens by immunization or natural colonization with the microbiota, influences the clonal distribution of antigen-specific innate-like B cells leading to quantitative increases in natural antibody (Nab) levels. These effects impact adult susceptibility to infection, and immune-responsiveness to allergens and auto antigens bearing similar B cell epitopes. Although the microbiota impacts the development of other lymphoid lineages its role in the development of the adult innate-like B cell and NAb repertoires is largely unexplored. Our hypothesis is that in addition to the well-described selection processes that occur during B cell development, the accessibility to autologous antigens and exogenous commensal-derived epitopes combine during neonatal development to shape the natural repertoire. The goal of these proposed studies is to elucidate mechanisms of exogenous antigen-directed development of the innate-like B cell repertoire using three model B cell antigen- specificities that represent epitopes for relevant pathogenic and commensal organisms with a spectrum of auto antigenic potential: phosphorylcholine (PC), N-acetyl-D-glucosamine (GlcNAc), and dextran (DEX). In Aim 1 of these studies wild-type gnotobiotic mice will be colonized with defined microbiota to assess the contribution of exogenous antigen signaling on clonal B cell development and formation of the NAb repertoire. Bone marrow chimera systems, composed of antigen-specific immunoglobulin (Ig) heavy chain-transgenic mouse strains, and lineage tracking will enable assessment of endogenous and exogenous antigen signaling on B lymphocyte selection and receptor editing processes. In Aim 2 we will analyze the frequency, cell surface phenotype, subset and isotype distribution as well as immunoglobulin gene usage of human B cells specific for these antigens. Examination of B cells isolated from cord blood, tonsils/adenoids, and peripheral blood B cells of donors of different ages will provide novel insight into the role of antigen experience in the formation of innate- like B cell repertoire diversity within and across individuals. Antigen-specific Ig genes will be cloned and expressed as recombinant antibody to determine effects of antigen-driven maturation on functions of these human antibodies. Collectively, these studies will define the microbial influences on NAb development in mice, determine developmental kinetics of human NAb development, and compare and contrast the NAb repertoire development between both species. This insight into the human infant responses to infection and colonization will advance our long-term goal of developing effective neonatal vaccine strategies and interventional therapies that provide protection from infectious pathogens and allergic and autoimmune diseases.

摘要 天然类B细胞衍生天然抗体(NAB)抗T非依赖性多糖(PS)和 磷脂表位在哺乳动物中普遍存在，并在保护人类免疫系统中发挥重要作用。 许多细菌病原体。然而，在感染或感染后，针对PS抗原的强大抗体反应 新生儿缺乏免疫接种。我们之前的研究表明，新生儿暴露在PS和磷脂中 通过免疫或与微生物区系自然定植的抗原，影响 抗原特异性先天B细胞导致天然抗体(NAB)水平的定量增加。这些 影响成人对感染的敏感性，以及对过敏原和自身抗原的免疫反应性 携带相似的B细胞表位。虽然微生物区系影响其他淋巴谱系的发展，但其 在成人先天类B细胞和NAB谱系的发育中的作用在很大程度上还没有被探索。我们的 假说是，除了在B细胞发育过程中发生的精心描述的选择过程之外， 新生儿对自体抗原和外源性共生抗原表位的可及性 发展以塑造自然曲目。这些拟议的研究的目标是阐明 利用三种模型B细胞抗原外源抗原诱导的先天类B细胞谱系的建立 代表相关病原和共生生物表位的特异性，具有AUT谱 抗原性：磷胆碱(PC)、N-乙酰-D-氨基葡萄糖(GlcNAc)和葡聚糖(DEX)。在目标1中 这些研究将用定义的微生物区系来定植野生型灵知生菌小鼠，以评估 外源抗原信号对克隆B细胞发育和NAB谱系形成的影响。骨髓 由抗原特异性免疫球蛋白(Ig)重链转基因小鼠品系组成的嵌合体系统， 血统追踪将使评估B淋巴细胞上的内源性和外源性抗原信号成为可能 选择和受体编辑过程。在目标2中，我们将分析频率、细胞表面表型、 人类B细胞亚群和亚型分布及免疫球蛋白基因利用 抗原。从脐带血、扁桃体/腺样体和外周血分离的B细胞的检测 不同年龄的捐赠者将提供新的见解，了解抗原经验在先天性心脏病形成中的作用。 就像B细胞系在个体内和个体间的多样性一样。将克隆抗原特异性免疫球蛋白基因并 表达为重组抗体以确定抗原驱动成熟对这些细胞功能的影响 人类抗体。总的来说，这些研究将确定微生物对小鼠NAB发育的影响， 测定人类NAB发育的发育动力学，并比较和对比NAB谱系 两个物种之间的发展。这种对人类婴儿对感染和殖民的反应的洞察 将推进我们开发有效的新生儿疫苗策略和介入疗法的长期目标 提供对传染性病原体以及过敏性和自身免疫性疾病的保护。

项目成果

Antibodies generated against Streptococci protect in a mouse model of disseminated aspergillosis.

产生的抗链球菌抗体可以保护播散性曲霉病小鼠模型。

• DOI: 10.4049/jimmunol.1401940
• 发表时间: 2015
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Wharton,RebekahE;Stefanov,EmilyK;King,RGlenn;Kearney,JohnF
• 通讯作者: Kearney,JohnF

Chemical Synthesis and Application of Biotinylated Oligo-α-(1 → 3)-d-Glucosides To Study the Antibody and Cytokine Response against the Cell Wall α-(1 → 3)-d-Glucan of Aspergillus fumigatus.

• DOI: 10.1021/acs.joc.8b01142
• 发表时间: 2018-11-02
• 期刊: The Journal of organic chemistry
• 作者: Komarova BS;Wong SSW;Orekhova MV;Tsvetkov YE;Krylov VB;Beauvais A;Bouchara JP;Kearney JF;Aimanianda V;Latgé JP;Nifantiev NE
• 通讯作者: Nifantiev NE