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: 10019973
• 负责人: Rachel R. Caspi
• 金额: $ 307.07万
• 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/10019973
• 关键词: Affect; Animal Model; Antibiotic Therapy; Area; Autoimmune Responses; Autoimmunity; Backcrossings; Binding; Bioinformatics; Birth; Brain; CD4 Positive T Lymphocytes; Cell physiology; Cells; Clinical; Collaborations; Corynebacterium; Data; Development; Diet; Disease; Ebola virus; Elements; Epitopes; Equilibrium; Eye; Genes; Genetic; Geographic Locations; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Holly; Homeostasis; Host Defense; IL17 gene; Immune; Immune Cell Activation; Immune response; Immunity; Immunization; Immunologics; Immunology; Immunotherapy; Infectious Agent; Inflammasome; Inflammation; Inflammatory Response; Innate Immune Response; Innate Immune System; Interferon Type II; Interferons; Interleukin-1; Interleukin-17; Intestines; Israel; Knock-in Mouse; Laboratories; Lead; Literature; Lymphocyte; Manuscripts; Measurable; Mediating; Modeling; Molecular; Mucosal Immune Responses; Mucous Membrane; Mus; Mutation; Nasopharynx; National Institute of Allergy and Infectious Disease; Natural Immunity; Oral cavity; Organism; Pathogenesis; Pathogenicity; Patients; Play; Predisposition; Preparation; Process; Production; Publishing; Regulation; Regulatory Pathway; Regulatory T-Lymphocyte; Research; Retina; Retinal; Role; Source; Surface; T cell response; T-Cell Receptor; T-Lymphocyte; TLR2 gene; Therapeutic; Transgenic Organisms; Treatment Efficacy; Tretinoin; Universities; Uveitis; Vagina; Viral; Vision; Vitamin A; Vitamins; Zika Virus; adaptive immunity; autoimmune uveitis; clinical material; commensal microbes; cytokine; effector T cell; gain of function mutation; gut microbiota; immunoregulation; interleukin-22; interstitial retinol-binding protein; intraepithelial; microbiome; microbiota; mouse model; mucosal site; neonatal infection; neutralizing antibody; novel; ocular surface; pathobiont; receptor; response; sensor

AUTOIMMUNE AND INFLAMMATORY RESPONSES AFFECTING THE RETINA Most of these studies emphasize mouse models of autoimmune uveitis developed in our laboratory, (i) experimental autoimmune uveitis (EAU) induced in mice by immunization with the retinal Ag IRBP, and (ii) spontaneous uveitis, which develops in R161H mice that express a transgenic T cell receptor (TCR) specific for the IRBP epitope encoded by residues 161-180, on the B10.RIII background. To better study basic mechanisms, we have backcrossed numerous gene-manipulated strains onto the B10.RIII/R161H background. (1) Recent findings in R161H mice indicated that gut commensal flora is necessary for development of spontaneous uveitis. Interestingly, while long term depletion of gut flora by antibiotic treatment, starting before birth, inhibits disease development in the spontaneous model, it had no effect on the (immunization induced) EAU model. However, a short-term antibiotic treatment did afford measurable protection. We found that long term antibiotic treatment depletes intraepithelial lymphocytes (IEL), which are known to be microbiota-dependent. These cells are able to inhibit activation of conventional lymphocytes, including CD4+ TCR uveitogenic T lymphocytes. We hypothesize that this may be connected to their regulatory function in uveitis, which is abrogated by long-term antibiotic treatment due to depletion of IEL. (Salvador et al., in preparation). (2) The role of various cytokines in pathogenesis and regulation of uveitis is being studied using both the classical and the spontaneous uveitis models, with emphasis on the balance between Th1 and Th17 responses, and their control by T-regulatory (Treg) cells. (a) Although previous studies indicated a critical role for the Th17 lineage cytokines in pathogenesis of immunization-induced EAU, crossing R161H mice to IFN-g deficient or IL-17 deficient mice revealed a major role for IFN- in the spontaneous disease. (b) Using mice deficient in both IL-17 and IFN-, we demonstrated that disease is still induced, but is now mediated by distinct effector cytokine profile. GM-CSF appears to be a critical pathogenic cytokine, which may drive disease in the absence of both IFN- and IL-17 (S Bing et al, manuscript in preparation). (c) On the other hand, IL-22 appears to have a protective role, which may at least in part be due to be due to a neuroprotective effect of IL-22. Data indicate that this is likely an indirect effect that requires other retinal cells (Mattapallil et al, J. Autoimmun 2019). (d) We reveal a novel regulatory pathway elicited by IL-17, a major proinflammatory cytokine. By binding to the IL-17R on activated T cells, IL17 elicits production of IL-24, which suppresses production of other Th17-related cytokines, including, importantly, GM-CSF. We speculate that the loss of this regulatory pathway in uveitis patients treated with anti-IL-17 neutralizing antibody (Secukinumab) may underlie the unexpectedly disappointing effects of this therapy (WP Chong et al., submitted). (e) In collaboration with Dr. Benjamin Sredni of Bar Ilan University, Israel, we showed that the immunomodulatory organotellurium compound, AS-101, ameliorates experimental autoimmune uveitis by regulating Th1 and Th17 responses and inducing Treg cells. (S Bing et al, J Autoimmun 2019). (3) 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 geographical regions where dietary VitA is limiting. (Horai, Zhou et al, in preparation). (4) In collaboration with Dr. Daniela Verthelyi and her group at FDA/CBER we are studying ocular effects of neonatal infection with Zika virus or Ebola virus constructs. The data suggest that immune privilege plays a role in promoting persistence of infectious agents within the CNS by protecting it from elimination by systemic immune mechanisms. These studies have the potential to help elucidate the mechanisms underlying the damage that these viral agents cause to the eyes and brain, and aid in the development of effective therapeutics (McWilliams et al., Cell Rep. 2019). EFFECTS OF INNATE IMMUNE RESPONSES ON OCULAR IMMUNITY AND AUTOIMMUNITY: Cellular and molecular elements of the innate immune system can affect immunopathogenic processes directly as well as indirectly, by affecting adaptive immunity. Innate immunity receptors have a major role in controlling susceptibility to autoimmune uveitis. A collaborative study with Dr. Holly Rosenzweig lab at OHSU, Portland, revealed that, unexpectedly, NOD2, usually thought of as an inflammation-promoting receptor, limits autoimmunity to the neuroretina. This appears to occur through a T cell intrinsic mechanism, by downregulating IL-17 production from T cells, but not through conventional APC priming. This study uncovers a hitherto unrecognized role for Nod2 as an inherent genetic modifier of T cell function in uveitis.. (Manuscript in revision). THE OCULAR SURFACE MICROBIOME AND MUCOSAL IMMUNE RESPONSES 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 V4 T cells respond to C. mast mainly via their TCR, whereas V6 T cells respond through innate receptors such as TLR2, and are highly dependent on IL-1. DC are involved in this response in a dual capacity as APC that present C. mast components in a C1d-dependent fashion, and as a source of IFN-. TLR2 appears to be required not only in T cells, but also in DC, for a maximal t cell IL-17 response. 2) A second direction of study examines commensal-elicited responses in an immunologically abnormal host, mouse and patient. 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 Gabriela Goldbach-Mansky who treats patients with NLRP3 inflammasome mutations, we have preliminary evidence to suggest that, in a host with an overactive inflammasome, an ocular surface commensal may elicit ocular surface inflammation, thus acting as a pathobiont. We are currently characterizing the local ocular immune response a in the mouse model and in patients with NLRP3 inflammasome mutation at the single-cell level.

自身免疫性和炎症反应影响视网膜 这些研究中的大多数都强调了在我们的实验室开发的自身免疫性葡萄膜炎的小鼠模型，（i）通过用视网膜AG IRBP免疫在小鼠中引起的实验性自身免疫性葡萄膜炎（EAU），以及（ii）自发性葡萄膜炎，在R161H小鼠中发育于R161H小鼠中，该R161H小鼠在R161H小鼠中表达了Transgenic T细胞受体（TCR）（TCR）的大量epp，irbp eppp eppp eppp eppp eppp eppp eppp eppp eppp eppp， B10.RIII背景。为了更好地研究基本机制，我们将众多基因操纵菌株反向B10.RIIII/R161H背景。 （1）R161H小鼠的最新发现表明，肠道菌群对于自发葡萄膜炎的发展是必需的。有趣的是，虽然从出生前开始，通过抗生素治疗对肠道菌群的长期消耗抑制了自发模型的疾病发展，但它对（免疫引起的）EAU模型没有影响。但是，短期抗生素治疗确实提供了可测量的保护。我们发现，长期的抗生素治疗耗尽了上皮淋巴细胞（IEL），已知是微生物群依赖性的。这些细胞能够抑制常规淋巴细胞的激活，包括CD4+ TCR uveitogenic T淋巴细胞。我们假设这可能与它们在葡萄膜炎中的调节功能有关，由于IEL耗尽而导致长期抗生素治疗消除。 （Salvador等人，在准备中）。 （2）使用经典和自发的葡萄膜炎模型研究了各种细胞因子在葡萄膜炎的发病机理和调节中的作用，重点是TH1和TH17反应之间的平衡，以及T-CRENULATIONT（TREG）细胞的控制。 （a）尽管以前的研究表明，Th17谱系细胞因子在免疫引起的EAU的发病机理中起着至关重要的作用，将R161H小鼠穿越IFN-G缺乏或IL-17缺乏小鼠，在自发性疾病中揭示了IFN的主要作用。 （b）使用缺乏IL-17和IFN-的小鼠，我们证明了疾病仍是诱导的，但现在是由独特的效应细胞因子谱介导的。 GM-CSF似乎是一种关键的致病细胞因子，在没有IFN-和IL-17的情况下可能会驱动疾病（S Bing等人，手稿中的手稿）。 （c）另一方面，IL-22似乎具有保护作用，至少部分是由于IL-22的神经保护作用。数据表明，这可能是需要其他视网膜细胞的间接效应（Mattapallil等，J。Autoimmun 2019）。 （d）我们揭示了由主要促炎细胞因子IL-17引起的新型调节途径。通过在活化的T细胞上与IL-17R结合，IL17引发了IL-24的产生，从而抑制了其他Th17相关细胞因子的产生，包括GM-CSF，包括GM-CSF。我们推测，用抗IL-17中和抗体（secukinumab）治疗的葡萄膜炎患者的这种调节途径的丧失可能是这种疗法意外令人失望的作用（WP Chong等人，已提交）。 （e）与以色列Bar Ilan大学的本杰明·萨里尼（Benjamin Sredni）合作，我们表明，免疫调节器官胞质化合物AS-101通过调节TH1和TH17反应并诱导Treg细胞来改善实验性自身免疫性葡萄膜炎。 （S Bing等人，J Autoimmun 2019）。 （3）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VAD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理区域具有临床意义。 （Horai，Zhou等人，准备）。 （4）与Daniela Verthelyi博士及其小组在FDA/CBER合作，我们正在研究使用Zika病毒或埃博拉病毒构建体的新生儿感染的眼部作用。数据表明，免疫特权在促进中枢神经系统内传染剂的持久性中发挥作用，通过保护其免受系统性免疫机制消除。这些研究有可能有助于阐明这些病毒剂对眼睛和大脑造成的损害的机制，并有助于开发有效的治疗疗法（McWilliams等人，CellRep。2019）。 先天免疫反应对眼部免疫和自身免疫的影响： 先天免疫系统的细胞和分子元素可以通过影响适应性免疫来直接以及间接影响免疫发作过程。 先天免疫受体在控制对自身免疫性葡萄膜炎的易感性方面具有重要作用。与波特兰OHSU的Holly Rosenzweig Lab博士进行的一项合作研究表明，出乎意料的是，NOD2通常被认为是促进症的受体，将自身免疫性限制为神经中的自身免疫性。这似乎是通过T细胞固有机制来实现的，通过下调T细胞的IL-17产生，而不是通过常规的APC启动。这项研究揭示了迄今无法识别的NOD2作为葡萄膜炎中T细胞功能的固有遗传修饰剂。（修订中的手稿）。 眼表面的眼表面微生物组和粘膜免疫反应 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，V4 T细胞主要通过其TCR对C.桅杆做出反应，而V6 T细胞通过先天受体（例如TLR2）反应，并且高度依赖于IL-1。 DC以双重能力为APC参与了这种响应，该响应以C1D依赖性的方式和IFN-来源。对于最大的T细胞IL-17响应，TLR2似乎不仅需要在T细胞中，而且在DC中都需要。 2）第二个研究方向研究了免疫学异常宿主，小鼠和患者的共生引起的反应。与沃伦·斯特罗伯（Warren Strober）博士（Niaid）合作，他们开发了表达NLRP3炎性症基因的功能性突变的敲门小鼠，而加布里埃拉·戈德巴赫·曼斯基（Gabriela Goldbach-Mansky）则与NLRP3炎性体突变的患者进行治疗，我们可以表现出炎症的效果，因此可以表现出炎症，因此可以表现出炎症的效果。作为病原体。我们目前正在表征小鼠模型中局部眼部免疫反应A和单细胞水平的NLRP3炎性体突变的患者中。