Mechanism of Humoral Immune Defects in Autoimmune Polyglandular Syndrome Type 1

自身免疫性多腺体综合征1型体液免疫缺陷的机制

• 批准号: 9304961
• 负责人: Kang Chen
• 金额: $ 19.25万
• 依托单位: WAYNE STATE UNIVERSITY
• 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 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9304961
• 关键词: Address; Antibodies; Antibody Formation; Antibody Repertoire; Antifungal Agents; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Avidity; B-Cell Activation; B-Lymphocytes; Biological Assay; Biology; Candida albicans; Candidiasis; Cell Communication; Cells; Cellular Immunity; Chemicals; Chronic Mucocutaneous Candidiasis; Clinical; Clinical Immunology; Complex; Cutaneous; Data; Defect; Disease; Dot Immunoblotting; Effector Cell; Endocrine Glands; Enzyme-Linked Immunosorbent Assay; Enzymes; Exhibits; Eye; Flow Cytometry; Gastrointestinal tract structure; Generations; Genes; Genetic Transcription; Genomics; German population; Goals; Health; Human; Immune; Immune System Diseases; Immune System and Related Disorders; Immune response; Immunity; Immunoglobulin Class Switching; Immunoglobulin G; Immunoglobulin Somatic Hypermutation; Immunoglobulin Switch Recombination; Immunologic Deficiency Syndromes; Immunologics; Impairment; In Vitro; Infection; Interleukin-17; Knowledge; Lymphoid; Lymphoid Tissue; Measures; Mediating; Methodology; Mission; Modeling; Mus; Mutation; Nuclear Localization Signal; Organ; Pathogenesis; Pathogenicity; Patients; Peripheral; Predisposition; Production; Public Health; Regulation; Research; Role; Side; Skin; Structure of germinal center of lymph node; Syndrome; T cell response; T-Lymphocyte; Testing; Therapeutic; Time; United States National Institutes of Health; Uracil; Work; acquired immunodeficiency; activation-induced cytidine deaminase; autoreactive B cell; autoreactivity; base; burden of illness; chromatin immunoprecipitation; congenital immunodeficiency; cytokine; disability; effective therapy; in vivo; innovation; insight; interleukin-22; loss of function mutation; mouse model; mutant; neutralizing antibody; next generation sequencing; novel; response; therapy development

Project Summary Autoimmune Polyglandular Syndrome type 1 (APS-1) is caused by mutations in the Autoimmune Regulator (Aire) gene, which normally promotes central and peripheral T cell tolerance. APS-1 patients exhibit autoimmune disorders in multiple organs and a predisposition to chronic mucocutaneous candidiasis (CMC). A lack of mechanistic under- standing of the basis of candidiasis and its paradoxical association with autoimmunity hampers effective treatment of APS-1. Patients with candidiasis often have defects in cell-mediated immunity. APS-1 patients produce class- switched IgG autoantibodies against T helper 17 (TH17) cytokines, which may neutralize and impair protective anti- fungal immunity. This highlights humoral immune dysfunction as a potentially important contributing factor to CMC in APS-1. Our long-term goal is to understand the immune dysregulation in autoimmune and immunodeficiency disor- ders and to develop mechanism-guided therapeutic strategies. The objective in this application is to elucidate the mechanism of humoral immune defects in APS-1. Our preliminary studies show that AIRE is expressed specifically in human and mouse germinal center (GC) B cells in secondary lymphoid organs in vivo, and in B cells undergoing antibody diversification in vitro. AIRE deficiency in B cells causes elevated class switch recombination (CSR) and somatic hypermutation (SHM). AIRE suppresses CSR when introduced into AIRE-deficient B cells. AIRE interacts in B cells with activation-induced cytidine deaminase (AID), the enzyme essential for SHM and CSR. Aire‒/‒ mice have aberrant expansion of GC B cell and T cell responses in systemic and mucosal lymphoid tissues. These findings support the central hypothesis that APS-1 involves aberrantly increased antibody maturation caused by B cell- intrinsic defects in AIRE-mediated AID regulation and B cell-extrinsic defects in the selection against autoreactive antibodies due to excessive GC response. Our rationale is that the elucidation of the mechanism of humoral immune defects in APS-1 will prompt more specific treatment of APS-1. The hypothesis will be tested by pursing two specific aims. Employing quantitative real-time PCR, flow cytometry, chromatin immunoprecipitation, genomic uracil dot blot and next-generation sequencing, studies in Aim 1 will determine the abnormalities in antibody maturation and the antibody repertoire in APS-1 and the mechanism by which AIRE regulates AID in B cells. Studies in Aim 2 will use competitive ELISA as well as mouse models of cutaneous Candida albicans infection to determine the role of B cell- intrinsic and -extrinsic AIRE deficiency in APS-1-associated candidiasis. This study proposes innovative and cutting- edge methodologies to test novel and mechanistic hypotheses in APS-1 pathogenesis. The work will have sustained and broad impact on the field of basic and clinical immunology, as it will not only advance the understanding of the disease APS-1 and AID-mediated regulation of antibody maturation, but also offer insights into the enigmas of an increasing number of immunodeficiencies that co-present with autoimmunity, thus paving the way to better treat the- se diseases.

项目概要 1 型自身免疫性多腺体综合征 (APS-1) 是由自身免疫调节因子 (Aire) 突变引起 基因，通常促进中枢和外周 T 细胞耐受，APS-1 患者表现出自身免疫性疾病。 多器官和慢性粘膜皮肤念珠菌病 (CMC) 的易感性缺乏机制不足。 念珠菌病的基础及其与自身免疫的矛盾关系阻碍了念珠菌病的有效治疗 APS-1。念珠菌病患者通常存在细胞介导的免疫缺陷。 将 IgG 自身抗体转换为针对 T 辅助细胞 17 (TH17) 细胞因子，这可能会中和并损害保护性抗- 这凸显了体液免疫功能障碍是导致 CMC 的潜在重要因素。 APS-1。我们的长期目标是了解自身免疫和免疫缺陷疾病中的免疫失调。 ders并开发机制引导的治疗策略。本申请的目的是阐明。 我们的初步研究表明AIRE在APS-1中特异性表达。 人和小鼠体内次级淋巴器官的生发中心 (GC) B 细胞，以及经历过 B 细胞中的 AIRE 缺陷会导致体外抗体多样化，从而导致类别转换重组 (CSR) 升高和 体细胞超突变（SHM）在引入 AIRE 缺陷的 B 细胞时会抑制 CSR。 具有激活诱导的胞苷脱氨酶 (AID) 的 B 细胞，这种酶是 SHM 和 Aire-/- 小鼠所必需的酶。 全身和粘膜淋巴组织中 GC B 细胞和 T 细胞反应的异常扩增。 支持APS-1涉及由B细胞引起的抗体成熟异常增加的中心假设 AIRE 介导的 AID 调节的内在缺陷和针对自身反应的选择中的 B 细胞外在缺陷 由于过度的GC反应而产生抗体我们的理由是阐明体液免疫的机制。 APS-1 的缺陷将促使对 APS-1 进行更具体的治疗。该假设将通过追求两个具体的结果来检验。 采用实时定量 PCR、流式细胞术、染色质免疫沉淀、基因组尿嘧啶斑点印迹。 和下一代测序，目标 1 的研究将确定抗体成熟的异常和 APS-1 中的抗体库以及 AIRE 调节 B 细胞中 AID 的机制将在 Aim 2 中的研究中使用。 竞争性 ELISA 以及皮肤白色念珠菌感染的小鼠模型，以确定 B 细胞的作用 APS-1 相关念珠菌病中的内在和外在 AIRE 缺陷本研究提出了创新和前沿的观点。 测试 APS-1 发病机制的新颖和机制假设的前沿方法这项工作将持续下去。 对基础和临床免疫学领域产生广泛影响，因为它不仅会增进对免疫学的理解 APS-1 和 AID 介导的抗体成熟调节，也提供了对疾病之谜的见解 与自身免疫共存的免疫缺陷数量不断增加，从而为更好地治疗以下疾病铺平了道路： 硒疾病。

项目成果

An alternatively transcribed TAZ variant negatively regulates JAK-STAT signaling.

替代转录的 TAZ 变体对 JAK-STAT 信号传导产生负向调节。

• 发表时间: 2019
• 影响因子: 7.7
• 作者: Fang, Chuantao;Li, Jian;Qi, Sixian;Lei, Yubin;Zeng, Yan;Yu, Pengcheng;Hu, Zhaolan;Zhou, Yufeng;Wang, Yulong;Dai, Ruping;Li, Jin;Huang, Shenglin;Xu, Pinglong;Chen, Kang;Ding, Chen;Yu, Fa
• 通讯作者: Yu, Fa

Stress Forces First Lineage Differentiation of Mouse Embryonic Stem Cells; Validation of a High-Throughput Screen for Toxicant Stress.

应激迫使小鼠胚胎干细胞首次谱系分化；

• DOI: 10.1089/scd.2018.0157
• 发表时间: 2019-01-11
• 期刊: Stem cells and development
• 影响因子: 4
• 作者: Quanwen Li;E. Louden;Jordan Z Zhou;S. Drewlo;Jing Dai;E. Puscheck;Kang Chen;D. Rappolee
• 通讯作者: D. Rappolee