Analysis of vascular Galectin-9 as an immunomodulator of B-cell activity

血管 Galectin-9 作为 B 细胞活性免疫调节剂的分析

• 批准号: 9807300
• 负责人: CHARLES J DIMITROFF
• 金额: $ 17.97万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9807300
• 关键词: Adhesions; Adhesives; Antibodies; Antibody Formation; Antibody Response; Antigens; Attenuated; Attenuated Vaccines; Autoimmune Diseases; Automobile Driving; B Cell Proliferation; B-Cell Activation; B-Lymphocyte Subsets; B-Lymphocytes; Binding; Biological Assay; Biology; Blood Vessels; CD22 gene; Carbohydrates; Cartoons; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cell model; Cell surface; Cells; Data; Development; Endothelial Cells; Endothelium; Foundations; Galactose Binding Lectin; Galactosides; Genes; Goals; Grant; High Endothelial Venule; Homeostasis; Homing; Human; Immune; Immune response; Immunity; Immunology; Immunomodulators; In Situ; Infection; Inhibition of Cell Proliferation; Investigation; Knowledge; Laboratories; Lead; Lectin; Leukocytes; Ligands; Lupus; Lymphoid; Mediator of activation protein; Memory; Memory B-Lymphocyte; Methods; Modeling; Molecular; Mus; Mutant Strains Mice; N-acetyllactosamine; Organ; PTPN6 gene; Peripheral; Physiological; Plasma Cells; Polysaccharides; Positioning Attribute; Proteins; Protocols documentation; Publishing; Receptors, Antigen, B-Cell; Regulation; Role; Signal Transduction; Structure; Structure of germinal center of lymph node; Supporting Cell; Therapeutic; Tissues; Tonsil; Vaccination; Vascular Endothelial Cell; blood group; cytokine; experience; high risk; immunoregulation; in vivo; innovation; insight; lymph nodes; mouse model; novel; novel therapeutics; peripheral tolerance; recruit; therapy design; transcriptome sequencing; Adhesions; Adhesives; Antibodies; Antibody Formation; Antibody Response; Antigens; Attenuated; Attenuated Vaccines; Autoimmune Diseases; Automobile Driving; B Cell Proliferation; B-Cell Activation; B-Lymphocyte Subsets; B-Lymphocytes; Binding; Biological Assay; Biology; Blood Vessels; CD22 gene; Carbohydrates; Cartoons; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cell model; Cell surface; Cells; Data; Development; Endothelial Cells; Endothelium; Foundations; Galactose Binding Lectin; Galactosides; Genes; Goals; Grant; High Endothelial Venule; Homeostasis; Homing; Human; Immune; Immune response; Immunity; Immunology; Immunomodulators; In Situ; Infection; Inhibition of Cell Proliferation; Investigation; Knowledge; Laboratories; Lead; Lectin; Leukocytes; Ligands; Lupus; Lymphoid; Mediator of activation protein; Memory; Memory B-Lymphocyte; Methods; Modeling; Molecular; Mus; Mutant Strains Mice; N-acetyllactosamine; Organ; PTPN6 gene; Peripheral; Physiological; Plasma Cells; Polysaccharides; Positioning Attribute; Proteins; Protocols documentation; Publishing; Receptors, Antigen, B-Cell; Regulation; Role; Signal Transduction; Structure; Structure of germinal center of lymph node; Supporting Cell; Therapeutic; Tissues; Tonsil; Vaccination; Vascular Endothelial Cell; blood group; cytokine; experience; high risk; immunoregulation; in vivo; innovation; insight; lymph nodes; mouse model; novel; novel therapeutics; peripheral tolerance; recruit; therapy design; transcriptome sequencing

PROJECT ABSTRACT Development of effective antibody (Ab) responses is critically dependent on the recruitment of naïve B cells into secondary lymphoid organs and, upon antigen activation, their coordinated differentiation into germinal center (GC) B cells to memory B cells and Ab-producing plasma cells. Knowing which mechanistic factors control B cell subset transition with the therapeutic goal of Ab-boosting or -attenuating activity, however, is still poorly understood. Exciting new published data from our laboratory highlight striking differences in the glycomic signatures of human naïve, GC and memory B cell subsets featured by either i-linear or I-branched poly-N-acetyllactosamines (poly-LacNAcs). Whereas GC B cells express mainly I-branched poly-LacNAcs, naïve/memory B cells display principally i-linear poly-LacNAcs, which enable robust binding to immuno- modulator, galectin (Gal)-9. Gal-9-binding causes downstream inhibition of cell proliferation, activation and signaling related to BCR-engagement while, interestingly, evoking a pro-survival activity. Given the selective Gal-9-binding to naive/memory B cells, we speculate that Gal-9 serves as a physiologic “tuner” of peripheral B cell activation and B cell reactivity. Other key data on the distribution of Gal-9 in “reactive” lymph node (LN)- like tonsil tissue reveal that, while naïve B cells express endogenous Gal-9, Gal-9 expression is strongest on high endothelial venules (HEV). Since HEVs initiate adhesive contact with circulating naïve/memory B cells and are densely packed in the cortex adjacent to B cell follicles, HEVs are strategically poised to elicit Gal-9- dependent adhesion/regulation. The spatial, cellular and functional control of Gal-9 on B cells in lymphoid organs is still unknown and a major gap in the field of galectin immunology. Our guiding hypothesis is that Gal- 9 on HEV can bind circulating naïve/memory B cells and help recruit them into LNs as well as transmit regulatory signals as B cells traverse the endothelium and enter B cell follicles. In this exploratory proposal, we will examine the function of human endothelial cell (EC)-derived Gal-9 on human naïve B cell adhesion and immunoregulation and will assay for B cell homing to peripheral LNs in the presence/absence of Gal-9. The Specific Aims are: 1.) To study Gal-9-dependent vascular EC – B cell adhesive interactions and 2.) To analyze immunoregulatory effects of Gal-9 expressed by cytokine-activated human ECs on human naïve B cells. We will employ our unique experience and innovative primary human cell models, adhesion assays, mutant mice, gene editing methods to study whether Gal-9 on ECs supports naïve B cell adhesion, LN-homing as well as trigger immunoregulatory activity. These exploratory studies directly challenge current dogma that galectins largely function as immunoregulators, offering an alternative role for Gal-9 in facilitating B cell-EC adhesion, LN-homing and in situ peripheral control of B cell activity. Importantly, our findings will reveal novel targets for modulating B cell immunity and provide necessary data for a grant larger in scope.

项目摘要 有效抗体（AB）反应的发展取决于幼稚B细胞的募集 进入次级淋巴机构，并在抗原激活后将其协调分化为生发 中心（GC）B细胞到记忆B细胞和产生AB的浆细胞。知道哪些机械因素 但是，控制B细胞子集的过渡具有促进或促进活动的治疗目标 理解不佳。令人兴奋的来自我们实验室的新发布的数据突出显示了 人类幼稚，GC和记忆B细胞子集的胶囊签名由i-linear或i分支 聚-N-乙酰甲胺（Poly-Lacnacs）。而GC B细胞主要表达I-Cranch的聚lacnacs，但 幼稚/记忆B细胞主要显示I线性聚lacnacs，这可以与免疫结合强大的结合 调节剂，半乳糖素（GAL）-9。 GAL-9结合引起下游抑制细胞增殖，激活和 有趣的是，与BCR的参与有关，有趣的是，引起了亲寿命活动的唤起。给定选择性 GAL-9结合到天真/记忆B细胞，我们推测GAL-9充当周围B的生理“调谐器” 细胞激活和B细胞反应性。关于“反应性”淋巴结（LN）中GAL-9分布的其他关键数据 - 像扁桃体组织一样，虽然幼稚的B细胞表达内源性Gal-9，但GAL-9表达在 高内皮静脉（HEV）。由于HEV启动与循环幼稚/记忆B细胞的粘合剂接触 并被荒漠地包装在与B细胞二元相邻的皮质中，HEV在战略上被毒化以引起Gal-9-- 依赖性的依从性/调节。淋巴样B细胞上GAL-9的空间，细胞和功能控制 器官仍然是未知的，并且是乳肠蛋白免疫学领域的主要差距。我们的指导假设是gal- 9在HEV上可以结合循环的幼稚/记忆B细胞，并有助于将其募集到LN中并发射 当B细胞横穿内皮并进入B细胞时，调节信号有效。在此探索性建议中 我们将检查人内皮细胞（EC）衍生的GAL-9对人幼稚B细胞粘附和 免疫调节，并将在存在/不存在GAL-9的情况下测定B细胞归巢对周围LN。这 具体目的是：1。）研究依赖GAL-9依赖性血管EC - B细胞粘合剂相互作用，并且2.） 分析细胞因子激活的人EC表达的GAL-9的免疫调节作用对人 幼稚的B细胞。我们将采用我们独特的经验和创新的原代人细胞模型，粘合剂 测定，突变小鼠，基因编辑方法，用于研究ECS上的Gal-9是否支持幼稚的B细胞粘附， LN-HOMing以及触发免疫调节活性。这些探索性研究直接挑战了当前 甲状腺素主要充当免疫调节剂的教条，为促进B提供了替代作用 细胞-EC粘附，LN植入和B细胞活性的原位外周控制。重要的是，我们的发现将 揭示了调节B细胞免疫力的新颖目标，并为范围较大的赠款提供必要的数据。