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.

总结 该项目的长期目标是深入了解抗体在健康和免疫中的性质和作用， 疾病B1细胞衍生的天然抗体在清除凋亡碎片中起关键的稳态作用 和有毒分子，超出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）可预防心脏病，而B2细胞- 衍生的IgG抗oxLDL会加剧心脏病。在人类中，IgM抗oxLDL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细胞不清楚。此外，病理性oxLDL代表了分子的异质性集合。 这些问题已经通过新的调查结果得到解决，为目前的提议奠定了基础。 1.我们将人类B1样细胞重新定义为CD20 + CD27 + CD43+，并通过多项标准显示它们是一种免疫缺陷细胞。 独特而独特的人口。2. Bengtsson及其同事将注意力集中在一种特定的肽上， 载脂蛋白B100（肽220），并显示血清抗氧化性IgM抗体水平， 修饰的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抗体是通过以下方式产生的： 人B1样细胞4. Bengtsson及其同事表明，血清IgG抗MGO-p220水平确实 与随后的心血管事件无关。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分支和动脉粥样硬化心脏之间的相互作用 疾病：为什么IgM抗体对oxLDL反对动脉粥样硬化，但IgG抗体对 oxLDL加剧动脉粥样硬化或不起作用。我们建议透过研究 IgM与IgG抗MG0-p220抗体的性质。为了阐明抗原特异性IgM 抗体是有益的，抗原特异性IgG抗体是无关紧要的，我们将：SA1。获得IgM 和IgG抗MGO-p220抗体进行结构比较，通过分选单细胞和PCR 扩增表达的免疫球蛋白用于序列分析。SA 2. Express原生和混合搭配 IgM与IgG抗体，通过将每种抗体克隆为IgM和IgG两者，并用和 没有J链。SA 3.检测IgM与IgG抗体的功能，通过检测 克隆/表达的抗体以抑制巨噬细胞摄取MGO修饰的apoB100。结果进行 研究将大大增加关于IgM天然抗体实现免疫应答的机制的知识。 在动脉粥样硬化和IgG抗体的有益作用不能做同样的。由此产生的信息 这项工作可能会提出新的治疗选择，包括管理，或免疫接种，具体 如本文所鉴定的抗体。