B Cell Autoimmunity in ApoE-deficient (ApoE-/-) Mice

ApoE 缺陷 (ApoE-/-) 小鼠的 B 细胞自身免疫

• 批准号: 39809166
• 负责人: Professor Dr. Andreas J.R. Habenicht
• 金额: --
• 依托单位: Institut für Prophylaxe und Epidemiologie der Kreislaufkrankheiten
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/39809166?language=en
• 关键词: Cell; Autoimmunity; ApoE; deficient; Mice

B lymphocyte subsets play opposing roles in atherosclerosis, yet it remains unclear where and how atherosclerosis B cell immunity is organized. In unpublished studies we explored B cell immunity in the arterial wall vs secondary lymphoid organs (SLOs) in mice. Transcript maps, flow cytometry, cell transfers, and immunoglobulin ELISPOT assays revealed that atherosclerosis B cell immunity is organized in artery tertiary lymphoid organs (ATLOs) during aging. Large adventitial B cell-related transcriptomes were identified in ATLOs vs intima plaques and SLOs. ATLO B-2 B cell subtypes included naïve, transitional, follicular, germinal centre, switched Ig+, IgG1+, IgA+, and IgE+ memory cells, B10+ B regulatory cells, and short- and long-lived plasma cells. Constitutively IgM- and IgG-secreting plasma cells were abundant in ATLOs though they were low in SLOs. Marked numbers of B-1 cells accumulated in ATLOs and this B cell compartment was strongly skewed towards B-1b versus B-1a cells. We conclude that ATLOs orchestrate multi-layered atherosclerosis B-1 and B-2 B cell responses during aging. The range of B-2 cell subtypes indicates autoantigen-triggered germinal center reactions, Ig class switching, and constitutive IgM and IgG secretion within diseased arterial wall segments. Our studies obtained during the past 2-3 years showed that we i) established methods to examine the BCR repertoires by NGS; ii) established the methods to obtain the paired H + L chains from single B cells. Using these technologies, our preliminary data suggest that autoimmune B cell responses may occur at different stages of atherosclerosis development in ApoE-/- mice. Germinal center B cells were isolated, their BCRs sequenced and cloned, and the respective Igs were in vitro expressed and the corresponding antibodies obtained. The first test of these antibodies shows specific binding to unknown structures in the adventitia and in the adjacent plaques though their reactivity with smooth muscle cells, endothelia cells, and adipocytes remained negative. In the current program, we aim at the isolation of the autoantigens using a variety of techniques, test the reactivity of ATLO-derived autoimmune antibodies, and begin to examine the impact of the antibodies for atherosclerosis progression in vivo.

B淋巴细胞亚群在动脉粥样硬化中起着相反的作用，但尚不清楚动脉粥样硬化B细胞免疫疾病的何处以及如何组织。在未发表的研究中，我们探讨了小鼠中B细胞免疫史壁与次要淋巴机器人（SLO）。转录图，流式细胞术，细胞转移和免疫球蛋白ELISPOT分析表明，在衰老过程中，动脉粥样硬化B细胞免疫史在第三纪淋巴机器人（ATLOS）中组织。在Atlos vs Intima斑块和SLO中鉴定出大型晚期B细胞相关的转录组。 ATLO B-2 B细胞亚型包括幼稚，过渡，卵泡，生发中心，开关IG+，IgG1+，IgA+和IgE+记忆细胞，B10+B调节细胞以及短寿命的浆细胞。宪法上的IgM和IgG分泌浆细胞在ATLOS中很丰富，尽管它们的SLOS较低。在ATLOS中积累的B-1细胞数量有明显的数量，该B细胞室与B-1B相对于B-1A细胞强烈偏斜。我们包括Atlos在衰老过程中协调多层动脉粥样硬化B-1和B-2 B细胞反应。 B-2细胞亚型的范围表明自动抗原触发的生发中心反应，IG类切换，以及疾病动脉壁段内的IgM和IgG分泌。在过去的2 - 3年中，我们获得的研究表明，我们建立了检查NGS BCR曲目的方法； ii）建立了从单个B细胞中获得配对的H + L链的方法。使用这些技术，我们的初步数据表明，自身免疫性B细胞反应可能发生在APOE - / - 小鼠的动脉粥样硬化发展阶段。分离生发中心B细胞，测序并克隆其BCR，并在体外表达相关的Ig，并获得相应的抗体。这些抗体的第一个测试表明，尽管它们与平滑肌细胞，内皮细胞和脂肪细胞的反应性，但它​​们与相邻的斑块中未知结构的特异性结合。在当前程序中，我们旨在使用各种技术分离自身抗原，测试ATLO衍生的自身免疫性抗体的反应性，并开始检查抗体对动脉粥样硬化进展的影响。