Immunization against Atherosclerosis

预防动脉粥样硬化的免疫

• 批准号: 8204868
• 负责人: AOSHUANG CHEN
• 金额: $ 19.63万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-15 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8204868
• 关键词: Accounting; Adjuvant; Adoptive Transfer; Animals; Antiatherogenic; Antigen Targeting; Antigens; Apolipoprotein E; Arteries; Atherosclerosis; Autoimmune Diabetes; B-Lymphocytes; Blood; Cell physiology; Cells; Cessation of life; Cholesterol; Chronic; Collagen; Cryoultramicrotomy; Data; Delayed Hypersensitivity; Developed Countries; Development; Dexamethasone; Disease; Equilibrium; Eragrostis; Flow Cytometry; Glucocorticoids; Grant; Heart Diseases; IL2RA gene; Immune; Immunity; Immunization; Immunologic Deficiency Syndromes; Immunosuppressive Agents; Inflammatory; Inflammatory Response; Insulin-Dependent Diabetes Mellitus; Intervention; Lead; Lesion; Microbe; Mus; Ovalbumin; Pathologic; Pathway interactions; Peptides; Play; Process; Regimen; Regulatory T-Lymphocyte; Role; Site; Smooth Muscle Myocytes; Specificity; Spleen; Stroke; Structure of brachiocephalic artery; T cell response; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Time; Toxic effect; aortic arch; atherogenesis; desensitization; design; enzyme linked immunospot assay; in vivo; insight; macrophage; male; mouse model; novel; peripheral blood; peripheral tolerance; protective efficacy; response; tumor

PROJECT SUMMARY Atherosclerosis is driven partially by pathogenic CD4+ effector T cells (Teff) reactive to antigens upregulated in the lesion ("athero-antigens"). Importantly, CD4+CD25+Foxp3+ regulatory T cells (Treg), a subset of T cells that function to suppress the response of Teff, have been shown to play a role in controlling atherosclerosis. Within the context of atherosclerosis an imbalance may exist between pathogenic Teff and suppressive Treg in response to athero-antigens, leading to the promotion of atherogenesis. These notions prompted us to design an anti-atherosclerosis strategy aimed at resetting the balance between Treg and Teff via increasing athero-specific Treg in vivo. Previously, mirroring the use of the conventional adjuvants for enhancing immunity, we explored the concept of "tolerogenic adjuvant" and designed a novel immunization strategy. In mouse models of delayed-type hypersensitivity (DTH) and spontaneous type I diabetes, we showed that immunization with peptide antigen, when performed under the influence of the immunosuppressant dexamethasone (DEX) serving as tolerogenic adjuvant (a strategy we termed "suppressed immunization"), caused long-term desensitization of DTH and blockade of autoimmune diabetes, respectively. We showed that such protective efficacy was associated with selective expansion of antigen-specific Treg. Encouraged by these results, we here will test our hypothesis that suppressed immunization (SI) using DEX and athero-antigens may selectively expand athero-specific Treg and reduce atherosclerosis. Under Specific Aim #1, we will determine whether SI increases the ratio of Treg to Teff at the lesion site and in peripheral blood in treated mice. To that end, male ApoE-/-/Foxp3-GFP+/+ mice (we have recently generated) will be treated with SI combining DEX with HSP60-derived peptide HP1. As controls, mice will be treated with PBS or HP1 alone. Innominate arteries from treated mice will be cryosectioned, and immunohistochemical analysis will be performed to quantify Teff (CD4+Foxp3-) and Treg (CD4+Foxp3-GFP+). Antigen specificity of Treg will be analyzed via ELISPOT with single cells prepared from the aortic arch. In addition, Teff and Treg in peripheral blood will be quantified via immunostaining and flow cytometry, and antigen specificity of the Treg will be assessed by their proliferation in response to the immunizing peptide HP1. Under Specific Aim #2, we will assess anti-atherogenic efficacy of SI in treated animals. To that end, male ApoE-/- mice with established atherosclerosis will be divided into 3 groups (n e 6). One group will be dissected before treatment to establish the baseline for lesion extent. Each of the 2 treatment groups will be treated with HP1 alone (control) or SI. Subsequently, lesions in the innominate artery will be quantified by histochemical/morphometric analysis. In addition, sections of the innominate artery will be analyzed for contents of macrophage, smooth muscle cell (SMC), and collagen. Statistical analyses will be performed on obtained data. Under Specific Aim #3, we will exploit adoptive transfer to determine whether SI-induced Treg contribute to the anti-atherogenic efficacy of SI. To that end, ApoE-/-/Foxp3-GFP+ mice will be treated with SI; 4 weeks later, Treg (Foxp3-GFP+) will be isolated from the spleen and used as donor Treg (test Treg). Treg isolated from non-treated mice will be used as control Treg. CD4+/Foxp3- cells isolated from ApoE-/- mice that have developed advanced atherosclerosis will be used as pathogenic Teff. Subsequently, pathogenic Teff will be adoptively transferred alone (control), or in combination (in various ratios) with test Treg or control Treg, into recipient ApoE-/-/Rag-2-/- mice (which are T cell- and B cell-deficient). Six weeks later, atherosclerosis lesion extent in recipients will be analyzed. Completion of these specific aims is likely to generate a safe, effective, and practical anti-atherosclerosis therapy. In addition, this study will further define the role of athero-specific Treg in controlling atherosclerosis and provide useful insights pertaining to targeting these Treg optimally.

项目概要 动脉粥样硬化部分是由对抗原产生反应的致病性 CD4+ 效应 T 细胞 (Teff) 驱动的 病变部位上调（“动脉粥样硬化抗原”）。重要的是，CD4+CD25+Foxp3+调节性T细胞（Treg）是一种 具有抑制 Teff 反应功能的 T 细胞亚群已被证明在控制 动脉粥样硬化。在动脉粥样硬化的背景下，致病性画眉草和 抑制性 Treg 响应动脉粥样硬化抗原，从而促进动脉粥样硬化形成。这些观念 促使我们设计一种抗动脉粥样硬化策略，旨在通过以下途径重置 Treg 和 Teff 之间的平衡 增加体内动脉粥样硬化特异性 Treg。此前，与传统佐剂的使用类似 增强免疫力，我们探索了“耐受佐剂”的概念，设计了一种新型免疫剂 战略。在迟发型超敏反应（DTH）和自发性 I 型糖尿病小鼠模型中，我们发现 在免疫抑制剂的影响下进行肽抗原免疫 地塞米松（DEX）作为耐受性佐剂（我们称之为“抑制免疫”的策略）， 分别引起DTH的长期脱敏和自身免疫性糖尿病的阻断。我们证明了 这种保护功效与抗原特异性 Treg 的选择性扩增有关。受到这些的鼓舞 结果，我们在这里将检验我们的假设，即使用 DEX 和动脉粥样硬化抗原抑制免疫 (SI) 可能 选择性地扩大动脉粥样硬化特异性Treg细胞并减少动脉粥样硬化。在具体目标#1下，我们将确定 SI 是否会增加治疗小鼠病变部位和外周血中 Treg 与 Teff 的比率。对此 最后，雄性 ApoE-/-/Foxp3-GFP+/+ 小鼠（我们最近生成的）将接受 SI 结合 DEX 的治疗 与 HSP60 衍生肽 HP1 一起。作为对照，小鼠将单独用 PBS 或 HP1 处理。无名动脉 来自治疗小鼠的细胞将被冷冻切片，并进行免疫组织化学分析以量化 Teff (CD4+Foxp3-) 和 Treg (CD4+Foxp3-GFP+)。 Treg 的抗原特异性将通过 ELISPOT 进行分析 从主动脉弓制备的单细胞。此外，外周血中的 Teff 和 Treg 将通过 免疫染色和流式细胞术，并通过 Treg 的增殖来评估其抗原特异性 响应免疫肽 HP1。根据具体目标#2，我们将评估以下药物的抗动脉粥样硬化功效： 治疗动物中的 SI。为此，将患有动脉粥样硬化的雄性 ApoE-/- 小鼠分为 3 类 组（n e 6）。治疗前将解剖一组以确定病变程度的基线。每一个 2 个治疗组将单独使用 HP1（对照）或 SI 进行治疗。随后，无名部位出现病变 动脉将通过组织化学/形态测量分析进行量化。此外，无名动脉的部分 将分析巨噬细胞、平滑肌细胞（SMC）和胶原蛋白的含量。统计分析将 对获得的数据进行。根据具体目标#3，我们将利用收养转移来确定是否 SI 诱导的 Treg 有助于 SI 的抗动脉粥样硬化功效。为此，ApoE-/-/Foxp3-GFP+ 小鼠将 接受 SI 治疗； 4周后，将从脾脏中分离出Treg（Foxp3-GFP+）并用作供体Treg （测试Treg）。从未治疗的小鼠中分离出的 Treg 将用作对照 Treg。 CD4+/Foxp3- 细胞分离自 已发展为晚期动脉粥样硬化的 ApoE-/- 小鼠将被用作致病性苔麸。随后， 致病性画眉草将单独（对照）或与测试组合（以不同比例）过继转移 Treg 或对照 Treg 进入受体 ApoE-/-/Rag-2-/- 小鼠（T 细胞和 B 细胞缺陷）。六周 随后，将分析受者的动脉粥样硬化病变程度。完成这些具体目标可能 产生安全、有效、实用的抗动脉粥样硬化疗法。此外，本研究将进一步明确 动脉粥样硬化特异性 Treg 在控制动脉粥样硬化中的作用并提供有关靶向的有用见解 这些 Treg 处于最佳状态。

项目成果

Dexamethasone promotes tolerance in vivo by enriching CD11clo CD40lo tolerogenic macrophages.

地塞米松通过富集 CD11clo CD40lo 耐受性巨噬细胞来促进体内耐受性。

• 发表时间: 2013-01
• 影响因子: 5.4
• 作者: Zheng, Guoxing;Zhong, Shibo;Geng, Yajun;Munirathinam, Gnanasekar;Cha, Isaac;Reardon, Catherine;Getz, Godfrey S;van Rooijen, Nico;Kang, Youmin;Wang, Bin;Chen, Aoshuang
• 通讯作者: Chen, Aoshuang

Complete Tolerogenic Adjuvant Stimulates Regulatory T Cell Response to Immunization.

完全耐受佐剂刺激调节性 T 细胞对免疫的反应。

• 发表时间: 2023-03-01
• 作者: Zheng, Guoxing;Geng, Yajun;Yan, Zhaoqi;Shin, Soo Min;Joshi, Kanak;Panicker, Anjali;Shankar, Archana;Elangovan, Ramya;Koehler, Jason;Gnanasekar, Varun;Gilles, Jessica Ann;Munirathinam, Gnanasekar;Chen, Aoshuang
• 通讯作者: Chen, Aoshuang

HMGB1-Neutralizing IgM Antibody Is a Normal Component of Blood Plasma.

HMGB1 中和 IgM 抗体是血浆的正常成分。

• 发表时间: 2020
• 作者: Geng, Yajun;Munirathinam, Gnanasekar;Palani, Sunil;Ross, Joseph E;Wang, Bin;Chen, Aoshuang;Zheng, Guoxing
• 通讯作者: Zheng, Guoxing

Palmitate-derivatized human IL-2: a potential anticancer immunotherapeutic of low systemic toxicity.

棕榈酸酯衍生的人 IL-2：一种潜在的低全身毒性的抗癌免疫疗法。

• 发表时间: 2013-03
• 作者: Chou, Sharon H;Shetty, Aditya V;Geng, Yajun;Xu, Lipeng;Munirathinam, Gnanasekar;Pipathsouk, Anne;Tan, Isaiah;Morris, Timothy;Wang, Bin;Chen, Aoshuang;Zheng, Guoxing
• 通讯作者: Zheng, Guoxing

Cutting edge: Dexamethasone potentiates the responses of both regulatory T cells and B-1 cells to antigen immunization in the ApoE(-/-) mouse model of atherosclerosis.

前沿技术：地塞米松可增强 ApoE(-/-) 小鼠动脉粥样硬化模型中调节性 T 细胞和 B-1 细胞对抗原免疫的反应。

• 发表时间: 2014-07-01
• 作者: Chen, Aoshuang;Geng, Yajun;Ke, Hanzhong;Constant, Laura;Yan, Zhaoqi;Pan, Yue;Lee, Patricia;Tan, Isaiah;Williams, Kurt;George, Samantha;Munirathinam, Gnanasekar;Reardon, Catherine A;Getz, Godfrey S;Wang, Bin;Zheng, Guoxing
• 通讯作者: Zheng, Guoxing