IgM vs IgG natural antibodies that bind pathogenic apolipoprotein B100

结合致病性载脂蛋白 B100 的 IgM 与 IgG 天然抗体

• 批准号: 9305007
• 负责人: THOMAS L ROTHSTEIN
• 金额: $ 22.65万
• 依托单位: WESTERN MICHIGAN UNIV SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9305007
• 关键词: Address; Antibodies; Antibody Formation; Antigens; Apolipoproteins; Apoptotic; Atherosclerosis; Attention; B-Lymphocytes; Binding; Cardiovascular Diseases; Cardiovascular system; Cells; Characteristics; Clinical; Cloning; Collaborations; Collection; Coronary heart disease; Data; Development; Disease; Elements; Event; Fatty Acids; Foundations; Glucose; Goals; Health; Heart Diseases; Housekeeping; Human; Immune system; Immunization; Immunoglobulin Constant Region; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Individual; Infection; J-Chain Immunoglobulins; Knowledge; Light; Lipoproteins; MS4A1 gene; Molecular; Mus; Nature; Oxides; Pathogenesis; Pathogenicity; Pathologic; Peptides; Play; Population; Prevention; Pyruvaldehyde; Reporting; Risk Factors; Role; Sequence Analysis; Serological; Serum; Sorting - Cell Movement; Speed; Stratification; Testing; Uncertainty; Variant; Work; aged; anti-IgG; anti-IgM; antigen binding; antimicrobial; arm; autoreactivity; base; cardiovascular risk factor; experience; experimental study; expression cloning; insight; macrophage; microbial; monomer; mouse model; novel therapeutics; oxidation; oxidized low density lipoprotein; peripheral blood; uptake

Summary The long term goal of this project is to gain insight into the nature and role of antibodies in health and disease. B1 cell-derived natural antibodies play a critical homeostatic role in eliminating apoptotic debris and noxious molecules, beyond their role in microbial defense. In mice, B1 cell-derived IgM against atherogenic oxidized low density lipoprotein (oxLDL) protects against heart disease, whereas B2 cell- derived IgG anti-oxLDL exacerbates heart disease. In humans, IgM anti-oxLDL antibodies correlate with atheroprotection, but these antibodies could not be connected to B1 cells, because the definition of human B1 cells was unclear. Moreover, pathological oxLDL represents a heterogeneous collection of molecules. These issues have been addressed with new findings that establish the foundation for the current proposal. 1. We redefined human B1-like cells as CD20+CD27+CD43+ and showed by multiple criteria they are a unique and distinct population. 2. Bengtsson and colleagues focused attention on a specific peptide of apolipoprotein B100 (peptide 220) and showed that the serum level of IgM antibodies against the oxidatively modified peptide is inversely and independently correlated with subsequent cardiovascular events. 3. We showed in collaboration with Bengtsson that beneficial IgM anti-MGO-p220 antibodies are generated by human B1-like cells. 4. Bengtsson and colleagues showed that the serum level of IgG anti-MGO-p220 does not correlate in any way with subsequent cardiovascular events. These results highlight an unresolved issue concerning interaction between the serological arm of the immune system and atherosclerotic heart disease: why do IgM antibodies against oxLDL oppose atherosclerosis but IgG antibodies against oxLDL exacerbate atherosclerosis or do nothing. We propose to address this issue by examining the nature of IgM vs IgG anti-MGO-p220 antibodies. To elucidate the reason why antigen-specific IgM antibodies are beneficial and antigen-specific IgG antibodies are inconsequential, we will: SA1. Obtain IgM and IgG anti-MGO-p220 antibodies for structural comparison, by sorting single cells and PCR amplifying expressed immunoglobulin for sequence analysis. SA2. Express native and mix-and-match IgM vs IgG antibodies, by cloning each antibody as both IgM and IgG, and expressing IgM with and without J chain. SA3. Test the function of IgM vs IgG antibodies, by examining the ability of cloned/expressed antibodies to inhibit macrophage uptake of MGO-modified apoB100. Results of these studies will greatly increase knowledge regarding the mechanism by which IgM natural antibodies fulfill a beneficial role in atherosclerosis and IgG antibodies fail to do the same. The information generated by this work is likely to suggest new therapeutic options that involve administration of, or immunization for, specific antibodies as identified herein.

摘要 这个项目的长期目标是深入了解抗体的性质和在健康和健康中的作用 疾病。B1细胞衍生的天然抗体在消除凋亡碎片方面发挥着关键的动态平衡作用 和有毒分子，超越了它们在微生物防御中的作用。在小鼠中，B1细胞来源的IgM抗 致动脉粥样硬化的氧化低密度脂蛋白(OxLDL)可以预防心脏病，而B2细胞- 衍生的免疫球蛋白抗氧化低密度脂蛋白加重心脏病。在人类中，抗oxLDL的IgM抗体与 动脉粥样硬化保护，但这些抗体不能连接到B1细胞，因为人类的定义 B1细胞的情况尚不清楚。此外，病理性oxLDL代表了一组不同种类的分子。 这些问题已通过新的调查结果得到解决，这些调查结果为目前的提案奠定了基础。 1.我们将人的类B1细胞重新定义为CD20+CD27+CD43+，并通过多种标准表明它们是一种 独特而独特的种群。2.Bengtsson和他的同事将注意力集中在一种特定的多肽上 载脂蛋白B100(肽220)，并显示血清中抗氧化性IgM抗体水平 修饰后的多肽与随后的心血管事件呈负相关且独立相关。3.我们 与Bengtsson合作显示，有益的IgM抗MGO-p220抗体是由 人的类B1细胞。4.Bengtsson和他的同事表明，血清中抗MGO-p220的免疫球蛋白水平 与随后的心血管事件没有任何关联。这些结果突出了一个尚未解决的问题 关于免疫系统的血清学臂与动脉粥样硬化心脏之间的相互作用 疾病：为什么抗oxLDL的IgM抗体反对动脉粥样硬化，而IgG抗体反对动脉粥样硬化 OxLDL要么加剧动脉粥样硬化，要么什么都不做。我们建议通过研究 Ig M与Ig G抗MGO-p220抗体的性质阐明抗原特异性免疫球蛋白M 抗体是有益的，而抗原特异性的抗体是无关紧要的，我们将：SA1。获取IgM 与Ig G抗MGO-p220抗体进行结构比较，通过分选单细胞和聚合酶链式反应 扩增表达的免疫球蛋白进行序列分析。SA2.表示原生和混合匹配 IGM与Ig G抗体的比较，方法是将每一种抗体克隆为Ig M和Ig G，并用和 没有J链。SA3.检测免疫球蛋白M抗体与免疫球蛋白G抗体的功能 克隆/表达抗体以抑制MGO修饰的apoB100的巨噬细胞摄取。这些措施的结果 研究将大大增加对IgM天然抗体实现的机制的了解 在动脉粥样硬化中的有益作用和免疫球蛋白抗体未能起到同样的作用。由此生成的信息 这项工作可能会提出新的治疗选择，包括对特定的疾病进行管理或免疫。 在此鉴定的抗体。