Collagen VI antibody reduces atherosclerosis by activating monocyte/macrophage polarization in ApoE-/- mice
Collagen VI antibody reduces atherosclerosis by activating monocyte/macrophage polarization in ApoE-/- mice
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DOI：
10.1016/j.intimp.2022.109100
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发表时间：
2022-08-03
影响因子：
5.6
通讯作者：
Zhao, Ming
中科院分区：
文献类型：
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作者：
Liu, Xianyan;Su, Jinyu;Zhao, Ming
动脉粥样硬化（AS）一直被认为是一种自身免疫性疾病。然而，针对AS的免疫治疗研究有限。我们之前发现AS患者血清中结合α 5和α 6链胶原VI （COL6A5或COL6A6）的IgG明显高于健康受试者，这里我们试图确定它们是否具有AS保护作用，并试图开发针对它们的人类抗体。用COL6A5或COL6A6免疫ApoE（-/-）小鼠，发现COL6A6具有抗动脉粥样硬化的保护性抗原。构建噬菌体人源单链抗体文库，获得col6a6特异性的人源单链抗体文库，并将其克隆到修饰后的pcDNA3载体中表达全长人源单链抗体。ApoE（-/-）小鼠饲喂高脂饮食（HFD） 20周，每周3次注射CVI单克隆抗体（mAb）或同型对照抗体，经主动脉油红O染色发现CVI单克隆抗体可使斑块面积减少45%。流式细胞术预测CVI单抗诱导单核/巨噬细胞从M1向M2极化。此外，通过Luminex实验，CVI mAb可诱导动物血清中促炎细胞因子mcp -1和IL-1 β降低，IL -4和IL-10水平升高。总的来说，我们发现了一种新的动脉粥样硬化相关抗原-胶原VI及其保护片段-胶原VI α 6链（COL6A6），并证明了人源化抗COL6A6抗体治疗在ApoE（-/-）小鼠动物模型中可逆转动脉粥样硬化，诱导单核细胞/巨噬细胞从M1向M2极化。
Atherosclerosis (AS) has been regarded as an autoimmune disease. However, studies on immunotherapy against AS are limited. We previously found that IgG in AS patients serum binding to alpha 5 and 6 chain of collagen VI (COL6A5 or COL6A6) was significantly higher than that in healthy subjects, here we tried to identify whether they are AS-protective, and tried to develop human antibodies against them. ApoE(-/-)mice were immunized with COL6A5 or COL6A6 and COL6A6 was found a protective antigen against atherosclerosis. A phage display human single-chain antibody (scFv) library was constructed and COL6A6-specific scFv was obtained, and cloned into a modified pcDNA3 vector to express full-length human antibodies. ApoE(-/-)mice were fed a high-fat diet (HFD) for 20 weeks and administered three weekly injections of CVI monoclonal antibody (mAb) or isotype control antibody, CVI mAb was found to be able to reduce plaque area by 45 % via aorta oil red O staining. Flowcytometry method predicted that CVI mAb induced monocyte/macrophage polarization from M1 to M2. Furthermore, CVI mAb induced decreases of pro-inflammatory cytokines of MCP-1and IL-1 beta, and increases of IL -4 and IL-10 levels in animal serum by using the Luminex assay. Overall, we found a novel atherosclero-related antigen - Collagen VI, and its protective fragment -Collagen VI alpha 6 chain (COL6A6) and proved that humanized antibody against COL6A6 therapy regresses atherosclerosis and induces monocyte/macrophage polarization from M1 to M2 in ApoE(-/-)mice animal model.