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.

概括 该项目的长期目标是深入了解抗体在健康和健康方面的性质和作用。 疾病。 B1 细胞衍生的天然抗体在消除细胞凋亡碎片方面发挥着关键的稳态作用 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 细胞衍生的 IgM 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，但这些抗体无法与 B1 细胞连接，因为人类的定义 B1细胞不清楚。此外，病理性 oxLDL 代表了分子的异质集合。 这些问题已通过新发现得到解决，为当前提案奠定了基础。 1. 我们将人类 B1 样细胞重新定义为 CD20+CD27+CD43+，并通过多种标准表明它们是 独特且独特的人口。 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）并显示抗氧化 IgM 抗体的血清水平 修饰肽与随后的心血管事件呈负相关且独立相关。 3.我们 与 Bengtsson 合作表明，有益的 IgM 抗 MGO-p220 抗体是通过 人类 B1 样细胞。 4. Bengtsson 及其同事表明，抗 MGO-p220 IgG 的血清水平与 与随后的心血管事件没有任何关联。这些结果凸显了一个尚未解决的问题 关于免疫系统的血清学分支与动脉粥样硬化心脏之间的相互作用 疾病：为什么 oxLDL 的 IgM 抗体可以对抗动脉粥样硬化，而 IgG 抗体则可以对抗动脉粥样硬化 oxLDL会加剧动脉粥样硬化或无任何作用。我们建议通过审查来解决这个问题 IgM 与 IgG 抗 MGO-p220 抗体的性质。阐明抗原特异性IgM产生的原因 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，通过单细胞分选和 PCR 进行结构比较 扩增表达的免疫球蛋白用于序列分析。 SA2。表达原生和混合搭配 IgM 与 IgG 抗体，通过将每种抗体克隆为 IgM 和 IgG，并用 和 表达 IgM 不带 J 链。 SA3。通过检查 IgM 与 IgG 抗体的功能来测试 IgM 与 IgG 抗体的功能 克隆/表达的抗体可抑制巨噬细胞摄取 MGO 修饰的 apoB100。这些结果 研究将极大地增加关于 IgM 天然抗体实现功能的机制的知识 在动脉粥样硬化中发挥有益作用，而 IgG 抗体则不能起到同样的作用。由此产生的信息 工作可能会提出新的治疗选择，其中涉及特定的治疗或免疫接种 如本文所鉴定的抗体。