Genetic, Cellular and Molecular Mechanisms in Autoimmunity to Retina

视网膜自身免疫的遗传、细胞和分子机制

• 批准号: 10706090
• 负责人: Rachel R. Caspi
• 金额: $ 393.82万
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706090
• 关键词: Adopted; Affect; Agonist; Animal Model; Anti-Inflammatory Agents; Antibiotics; Arrestins; Aryl Hydrocarbon Receptor; Autoimmune Responses; Autoimmunity; Bioinformatics; Brain; Bruch' s basal membrane structure; COVID-19; COVID-19 patient; Cataract; Cells; Central Nervous System Diseases; Characteristics; Choroid; Clinic; Clinical; Collaborations; Corynebacterium; Crystalline Lens; Data; Development; Disease; Engineering; Equilibrium; Experimental Autoimmune Encephalomyelitis; Experimental Models; Extracellular Matrix; Eye; FOXP3 gene; Genes; Genetic; Germ-Free; Goals; Growth Factor; Homeostasis; Host Defense; Human; Immune; Immune response; Immunity; Immunization; Immunologics; Immunology; Immunotherapy; In Vitro; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Interleukin-1; Interleukin-17; Intervention; Intestines; Invaded; Knock-in Mouse; Laboratories; Lead; Leg; Legal; Leukocytes; Licensing; Literature; Lung; Lymphocyte; Lymphocyte Depletion; Measurable; Microglia; Molecular; Molecular Mimicry; Morbidity - disease rate; Mucosal Immune Responses; Mucous Membrane; Mus; Mutation; Nasopharynx; National Institute of Allergy and Infectious Disease; Neurons; Oral cavity; Organism; Participant; Pathogenesis; Pathogenicity; Patients; Peptides; Peripheral; Play; Predisposition; Preparation; Process; Publishing; Regulatory T-Lymphocyte; Retina; Role; Site; Spleen; Stimulus; Surface; T cell response; T-Cell Receptor; T-Lymphocyte; TLR2 gene; Therapeutic; Tissues; Transgenic Organisms; Tretinoin; United States National Institutes of Health; United States National Library of Medicine; Uveitis; Vagina; Virus; Vision; Vitamin A; Vitamins; Washington; autoimmune uveitis; autoinflammatory; clinical material; commensal microbes; cytokine; cytokine release syndrome; dietary; effector T cell; gain of function mutation; genome sequencing; gut microbes; gut microbiome; gut microbiota; human microbiota; humanized mouse; immune activation; interstitial retinol-binding protein; intraepithelial; lens capsule; microbial; microbiome; microbiota; mortality; mouse model; mucosal site; mutant; neuroinflammation; ocular surface; pathobiont; preservation; protective effect; receptor; reconstitution; resident commensals; response; sensor; transcriptome; whole genome

IMMUNITY AND AUTOIMMUNITY INVOLVING THE NEUORORETINA AND THE ROLE OF THE GUT MICROBIOME For most of these studies we utilize mouse models of autoimmune uveitis developed in our laboratory, (i) experimental autoimmune uveitis (EAU) induced in mice by immunization with the retinal antigen (Ag) IRBP, and (ii) spontaneous uveitis in R161H mice engineered to express a transgenic T cell receptor (TCR) specific for IRBP peptide 161-180. 1) Vitamin (VitA) derivatives are necessary for functional activation of immune cells (published literature). We previously demonstrated the importance of Vitamin A (VitA) and its metabolite, retinoic acid, in ocular immune privilege. Using mice made VitA deficient (VAD), we found that T cell effector function that was acquired before onset of VAD, is maintained in the VAD host. These findings may have clinical implications in regions where dietary VitA is limiting. (Horai, Zhou et al, in preparation). 2) Laquinimod is an aryl hydrocarbon receptor (AHR) agonist. We found that it has strong protective effects in the induced model of EAU by modulating multiple inflammatory and anti-inflammatory cytokines, and may affect composition of gut flora, which in turn affects development of disease. (Xu et al, in preparation) 3) "Licensing" of T cells for pathogenicity: It has been demonstrated that activated T cells specific to retina or brain infused into a new host first settle in peripheral tissues (spleen, lung) where they become "licensed" to invade the target tissue. We are currently examining whether the gut, which affects uveitis via its resident commensal flora, can serve as a licensing site that might be important in pathogenesis (Salvador et al., in preparation). 4) The role of IL-34 and microglia in EAU: IL-34 is a cytokine produced by neuronal tissue, which is a necessary growth factor for microglia, an immune-like call resident in the retina Preliminary data indicate that IL-34 depletion in the retina has a dampening effect on intraocular inflammation, suggesting that microglial cells may be active participants in the inflammatory process. (Peng, Mattapallil et al., in preparation) 5) Ocular immune privilege: We have previously shown that nave retina-specific T cells in the eye are primed locally, and after 7 days up to 50% convert into Foxp3+ T-regulatory cells (Tregs). We are currently examining the transcriptome of eye-induced Tregs and how it differs from Tregs primed at other tissue sites or in vitro, (Peng et al., in preparation). In collaboration with the Lin laboratory at Cleveland Clinic, we demonstrated that during uveitis, inflammatory cells are able to transmigrate between the choroid and retina through the RPE cell layer, which was not thought to occur, due to the presence of Bruchs membrane (ref.5). 6) Commensal flora is necessary for development of spontaneous uveitis, in part due to molecular mimicry of IRBP161-180. However, innate mechanisms also play a role, as evidenced by the finding that antibiotics (ABX) starting a week before immunization for EAU (where the antigenic stimulus is provided), afforded measurable protection. That protection, however, was temporary and disappeared on prolonged ABX. Results suggest that prolonged ABX depletes intraepithelial lymphocytes (IEL), which are known to be microbiota-dependent, We hypothesize that IEL may have an inhibitory role in uveitis, an effect which is reversed after long-term ABX due to IEL depletion. (Salvador R et al., in preparation). 7) Human gut flora and uveitis: We reconstituted germ free R161H mice with gut microbes from 3 healthy human donors and established 3 humanized flora mouse lines Results showed that human gut flora supports development of spontaneous uveitis. Mice adopted only a subset of the human flora, but preserved the microbial footprint of the donor. We are currently identifying microbial taxa associated with low and high disease scores, with the aim of developing interventional strategies as an approach to regulating uveitis through directed manipulation of the microbiome (Horai, Zhou, Murphy et al, in preparation). 8) in collaboration with Drs. Sue Menko (Thomas Jefferson Univ) and Mary Ann Stepp (George Washington Univ) we uncovered that during uveitis immune cells invade the lens of the eye through the extracellular matrix of the lens capsule. The results may help explain inflammation-induced cataract formation (ref.2) 9) Examined the consequence of the Alpk1 mutation on susceptibility to EAU, since this mutation results in autoinflammatory disease, including uveitis, in humans. Demonstrated that the human mutant alpk1 knock-in mice, otherwise unperturbed, do not develop spontaneous ocular inflammation (ref.3) 10) In collaboration with the Egwuagu lab at the NEI, LI, demonstrated that IL-27-producing B-1a cells (Bregs) suppress EAU and EAE, indicating a role in neuroinflammation and CNS disease in a broader sense (ref.1). MUCOSAL IMMUNE RESPONSES TO COMMENSALS AT THE OCULAR SURFACE Mucosal sites such as the intestine, oral cavity, nasopharynx, and vagina all have associated commensal flora. The surface of the eye is also a mucosal site, but presence of ocular surface microbiome was contentious. Previously, we isolated and purified a candidate ocular commensal, Corynebacterium mastitidis (C. mast). This organism elicits a commensal-specific IL-17 response from T cells in the ocular mucosa, tuning local host defense to afford protection from infectious pathogenic organisms. 1) We are examining the molecular sensors of C. mast in T cells. Data suggest that Vg4 T cells respond to C. mast mainly via their TCR, whereas Vg6 T cells respond through innate receptors such as TLR2, and are highly dependent on IL-1. TLR2 appears to be required not only in DC (where it contributes to IL-1 induction), but also in T cells. The mechanism of intrinsic T cell requirement for TLR2 is being investigated (Zhu & Xu et al. and Xu et al., in preparation). 2) We also examined commensal-elicited ocular surface responses in an immunologically abnormal host, mouse as well as human. In collaboration with the group of Dr. Warren Strober (NIAID) who developed knock-in mice expressing a gain-of-function mutation in the NLRP3 inflammasome gene, and Dr. Raphaela Goldbach-Mansky who treats patients with NLRP3 inflammasome related diseases, we obtained evidence to suggest that an ocular surface commensal can elicit ocular surface inflammation in an immunologically perturbed host, thus acting as a pathobiont. We are characterizing the local ocular immune response and the ocular surface microbiome in the mouse model and in patients with NLRP3 inflammasome mutations at the single-cell level (Siak, Mattapallil, St. Leger et al., in preparation). This will have implications for understanding and treatment of the characteristic ocular inflammation in patients with NLRP3-related diseases. 3) We performed whole genome sequencing, assembly and annotation of C. mast. https://dataview.ncbi.nlm.nih.gov/object/PRJNA758739?reviewer=dhmcn2fm0ngfj6bkh5jsq3u4d5 (Nagarajan et al., (ref4). COVID-19 related: EFFECTS OF THE MICROBIOME ON CYTOKINE STORM IN A "HUMANIZED" MOUSE MODEL. COVID-19 patients' morbidity and mortality is being attributed largely to elicitation of a cytokine storm due to an excessive immune response to the virus. The project uses immunodeficient NSG mice reconstituted with human leukocytes developed by Dr. Richard Flavell in collaboration with Regeneron, Inc. to study the effects of intestinal microbiome on cytokine storm. This project is currently stalled due to the inability of the NIH tech transfer to overcome legal barriers to obtaining these mice.

免疫力和自身免疫性涉及neuororetina和肠道微生物组的作用 对于大多数研究，我们利用了在我们的实验室中开发的自身免疫性葡萄膜炎的小鼠模型，（i）通过用视网膜抗原（AG）IRBP（AG）IRBP和（II）在R161H的R161H小鼠中以161 cr pexter（T）的特殊性（TCR）诱导的小鼠诱导的实验性自身免疫性葡萄膜炎（EAU）。 1）维生素（VITA）衍生物对于免疫细胞的功能激活是必需的（已发表的文献）。我们以前证明了维生素A（VITA）及其代谢产酸在眼部免疫特权中的重要性。使用使VITA缺乏（VAD）的小鼠，我们发现在VAD宿主中维持了VAD发作之前获得的T细胞效应函数。这些发现可能在饮食中限制的地区具有临床意义。 （Horai，Zhou等人，准备）。 2）拉奎尼莫德是一种芳基烃受体（AHR）激动剂。我们发现，它通过调节多种炎症和抗炎细胞因子而在EAU的诱导模型中具有强大的保护作用，并可能影响肠道菌群的组成，进而影响疾病的发展。 （Xu等，准备） 3）T细胞的致病性“许可”：已证明激活的T细胞被特异性的视网膜或脑注入了新宿主中，首先在周围组织（脾脏，肺）中降低了它们“获得许可”以侵入目标组织。我们目前正在研究通过其居民Consensal Flora影响葡萄膜炎的肠道是否可以用作一个可能在发病机理中很重要的许可部位（Salvador等人，在准备中）。 4）IL-34和小胶质细胞在EAU：IL-34中的作用是由神经元组织产生的细胞因子，神经元组织是小胶质细胞的必要生长因子，这是一种免疫呼叫，在视网膜的初步数据中驻留在视网膜的初步数据中，这表明IL-34在视网膜中可能会引起静脉内的炎症，这可能会引起静脉内的影响，这是在脑中的炎症，该症状是在脑上的易位，从而有效地发炎，从而有效地发挥了作用。 过程。 （Peng，Mattapallil等人，在准备中） 5）眼部免疫特权：我们先前表明，眼睛中的视网膜特异性T细胞是局部底漆的，在7天后，高达50％转化为Foxp3+ T调节细胞（Tregs）。我们目前正在研究眼睛诱导的Treg的转录组，以及它与在其他组织部位或体外启动的Treg的不同（Peng等人，在制备中）。与克利夫兰诊所的LIN实验室合作，我们证明了在葡萄膜炎期间，炎症细胞能够通过RPE细胞层在脉络膜和视网膜之间进行移民，这是由于BRUCHS膜的存在而被认为不会发生的（Ref.5）。 6）对于自发葡萄膜炎的发展是必要的，部分原因是IRBP161-180的分子模仿。然而，先天机制也起着作用，这一点可以从对EAU免疫（提供抗原刺激的地方）开始前一周开始，从而获得了可测量的保护。但是，这种保护是暂时的，并且在延长的ABX上消失了。结果表明，延长的ABX耗尽了上皮内淋巴细胞（IEL），已知是微生物群依赖性的，我们假设IEL可能在葡萄炎中具有抑制作用，这一作用是由于IEL DEPTELITION引起的长期ABX后逆转的作用。 （在准备中，Salvador R等人）。 7）人类肠道菌群和葡萄膜炎：我们用来自3个健康的人类供体的肠道微生物重构无细菌R161H小鼠，并建立了3种人源化的植物群小鼠线结果，表明人类肠道菌群支持自发性葡萄膜炎的发展。小鼠仅采用了人类植物的一部分，但保留了捐赠者的微生物足迹。我们目前正在确定与低疾病评分相关的微生物分类单元，目的是开发介入策略，作为通过定向操纵微生物组来调节葡萄膜炎的方法（Horai，hou，Zhou，Murphy等人，准备制备）。 8）与Drs合作。 Sue Menko（Thomas Jefferson Univ）和Mary Ann Stepp（George Washington Univ）我们发现，在葡萄膜炎免疫细胞中，通过镜头胶囊的细胞外基质侵袭了眼睛的镜头。结果可能有助于解释炎症引起的白内障形成（参考文献2） 9）检查了ALPK1突变对EAU敏感性的后果，因为这种突变导致人类中的自发性疾病，包括葡萄膜炎。证明人突变体Alpk1敲击小鼠否则不受干扰，不会出现自发的眼部炎症（参考文献3） 10）与NEI的Egwuagu实验室合作，LI，证明IL-27产生的B-1A细胞（BREGS）抑制了EAU和EAE，表明在更广泛的意义上在神经炎症和CNS疾病中起作用（Ref.1）。 粘膜免疫反应对眼表的共生反应 粘膜位点，例如肠道，口腔，鼻咽和阴道，均具有相关菌群。眼表面也是粘膜部位，但是眼表微生物组的存在是有争议的。以前，我们分离并纯化了候选眼球菌群mastitidis（C.桅杆）。这种有机体引起了眼粘膜中T细胞的共生特异性IL-17反应，调整了当地宿主防御，以保护免受感染性致病生物的保护。 1）我们正在检查T细胞中肥大梭状芽胞杆菌的分子传感器。数据表明，VG4 T细胞主要通过其TCR对C.桅杆做出反应，而VG6 T细胞通过先天受体（例如TLR2）反应，并且高度依赖IL-1。 TLR2不仅在DC（有助于IL-1诱导的地方），而且在T细胞中也需要。正在研究TLR2的内在T细胞需求的机制（Zhu＆Xu等人和Xu等人，在制备中）。 2）我们还检查了免疫学异常宿主，小鼠和人类中的共生引起的眼表面反应。 In collaboration with the group of Dr. Warren Strober (NIAID) who developed knock-in mice expressing a gain-of-function mutation in the NLRP3 inflammasome gene, and Dr. Raphaela Goldbach-Mansky who treats patients with NLRP3 inflammasome related diseases, we obtained evidence to suggest that an ocular surface commensal can elicit ocular surface inflammation in an immunologically perturbed host, thus acting as a Pathobiont。我们正在表征小鼠模型中的局部眼部免疫反应和眼表面微生物组，以及在单细胞水平下NLRP3炎性体突变的患者（Siak，Mattapallil，St.Leger等人，准备中）。这将对NLRP3相关疾病患者的特征性眼部炎症的理解和治疗产生影响。 3）我们进行了整个基因组测序，C。Mast的组装和注释。 https://dataview.ncbi.nlm.nih.gov/object/prjna758739?reviewer = dhmcn2fm0ngfm0ngfj6bkh5jsq3u4d5（nagarajan et al。，（ref4）。 COVID-19相关：微生物组对“人性化”小鼠模型中细胞因子风暴的影响。 COVID-19患者的发病率和死亡率主要归因于由于对病毒的过度免疫反应而引起的细胞因子风暴。该项目使用与理查德·弗拉维尔（Richard Flavell）博士与Regeneron，Inc。合作开发的人类白细胞重构的免疫缺陷的NSG小鼠，以研究肠道微生物组对细胞因子风暴的影响。由于NIH技术转移无法克服获得这些小鼠的法律障碍，目前该项目目前停滞不前。