Immune Responses To Ocular Antigens

对眼抗原的免疫反应

• 批准号: 8556801
• 负责人: Igal Gery
• 金额: $ 118.83万
• 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/8556801
• 关键词: Affect; Animals; Antibodies; Antigen Targeting; Antigen-Presenting Cells; Antigens; Autoantigens; Autoimmune Diseases; Autoimmunity; Biological; Blindness; CD28 gene; CD3 Antigens; CD4 Positive T Lymphocytes; Cell Line; Cells; Data; Development; Disease; Dissection; Environment; Etiology; Exhibits; Eye; Eye diseases; Gene Expression Profile; Helper-Inducer T-Lymphocyte; Immune; Immune response; Inferior; Inflammation; Inflammatory; Knockout Mice; Learning; Lymphocyte; Mediating; Microarray Analysis; Molecular Profiling; Monitor; Mus; Phenotype; Physiological; Physiological Processes; Play; Population; Predisposition; Preparation; Procedures; Process; Proliferating; Proteins; Publications; Regulatory T-Lymphocyte; Research; Resistance; Resolution; Role; Severities; Signal Transduction; Specificity; Spleen; Surface Antigens; System; T-Lymphocyte; TYROBP gene; Testing; Th1 Cells; Transgenic Mice; animal facility; chemokine; chemokine receptor; cytokine; in vivo; microbial; novel strategies; polarized cell; response

Immune-mediated inflammation, a major cause of vision loss, is not fully understood. This project is aimed at collecting new information concerning the mechanisms, cells and molecules involved in the pathogenic process of ocular inflammation. The collected information is to be used for development of new approaches for treatment of this group of eye diseases. Research in FY2012 focused mainly on three issues: (i) analysis of dissimilarities between T-helper (Th) cell lines generated by either the commonly used activation with anti-CD3/CD28 antibodies, or with the physiological activation mode by the specific antigen presented by antigen-presenting cells (APC); (ii) examining the features of natural- and induced-T-regulatory cells (nTreg and iTreg, respectively) with specificity toward an ocular antigen; (iii) further dissection of the surprising dual effect of DAP12 on development of EAU and related immune responses. Summary Targeted at learning about pathogenic processes of inflammatory eye diseases, this project focused in FY 2012 mainly on populations of T-cells involved in these processes. Specifically, we extended our study in FY 2012 by focusing on the following issues: (1) Dissimilarities between lines of Th cells generated by activation with anti-CD3/CD28 antibodies, or by the specific antigen presented by APC; (2) collecting information on different subpopulations of Treg cells; (3) further dissecting the surprising dual effect of DAP12 on immunopathogenic responses. 1) Dissimilarities between Th lineages generated by different mechanisms. Lineages of Th are generated by activation of nave CD4 cells concurrently with polarization by the lineage-specific cytokine(s). It is assumed that in vivo, the activation is provided by interaction of the nave CD4 cells with their specific Ag, presented by APCs. Since only miniscule proportions of CD4 cells with specificity toward tested Ags exist in preparations of CD4 cells from WT animals, the activation by Ag and APC has been commonly replaced by exposure of the CD4 cells to antibodies against two cellular molecules that participate in the physiological process of activation, namely, CD3 and CD28. The availability of TCR transgenic (Tg) mice, in which the majority of T-cells recognize the same antigen, made it possible, however, to generate lines of polarized Th cells by activation of nave CD4 cells by either the specific antigen presented by APC, or by the anti-CD3/CD28 antibodies. These transgenic mice made it possible to compare between lines of polarized cells generated by either one of the two modes of activation. We compared between lineages of Th1, Th9 and Th17, generated by either one of the activation modes, and found remarkable differences between the two lineages of each of the three phenotypes in the following biological activities: capacities to migrate to and proliferate in the recipient spleen and, importantly, to induce inflammation in the recipient mouse eyes expressing the target antigen; the expression profiles of certain chemokines, chemokine receptors and major surface antigens; and, notably, the pattern of gene expression, as monitored by microarray analysis. Our findings thus underscore the limitations of biological observations made with Th lineages generated by only activation of CD4 cells with the anti-CD3/CD28 Abs. 2) Differences among T-regulatory (Treg) populations generated by different modes. Specific Tregs, that inhibit immune response against self antigens, have the potential of being used to specifically inhibit pathogenic immune responses and are the focus of intensive research effort. The induction of Treg requires the cells to be activated during the process of polarization toward acquisition of the immune suppressive capacity. The activation process used in the majority of studies have been by antibodies against CD3/CD28, but other studies have employed specific activation by the antigen presented by APC. To our knowledge, no publication has compared Treg cells generated by these two procedures. Using our system of TCR transgenic mice (see above) made it possible to make such a comparison. In general, Treg cells induced by antigen and APC were more suppressive than those induced by activation with the antibodies. The inhibitory effects of the two lineages of Tregs were tested in cultures of nave CD4 during acquisition of polarity or in cultures of polarized Th1, Th9 or Th17 cells responding to the specific antigen (HEL). In all systems, Th17 exhibited the highest level of susceptibility, whereas Th1 cells were the most resistant and Th9 cells were intermediate in their susceptibility. 3) Resolution to the enigmatic dual effects of DAP-12 on immunopathogenic responses. DAP-12 (DNAX-activating protein of 12kDa) plays a major role in the immune response by transducing a variety of activation signals. The role of DAP-12 in the etiology of pathogenic autoimmunity has been controversial by the molecules capacity to either enhance or suppress experimental autoimmune diseases in different studies. In the present study we compared DAP-12 null mice and wild type (WT) controls for their susceptibility to EAU induction and immune responsiveness. Mice were reared in two animal facilities, with low (A) or high (B) standards. Although disease development in WT control mice from both facilities was comparable, DAP-12 null mice from facility A were superior to their WT controls in both the severity of their ocular inflammation and the levels of proliferation and cytokines produced by their lymphocytes. In contrast, DAP-12 null mice from facility B were inferior to their controls by both parameters. Our data thus provide a possible resolution to the controversy concerning the role of DAP-12: deficiency in this molecule can either promote or suppress the pathogenic response, depending on the environment. Further, we suggest that differences between the environments of the two animal facilities affected the immune responsiveness by modifying the microbial flora of the mice and consequently, their immune capacity.

免疫介导的炎症是视力丧失的主要原因，尚未完全了解。该项目旨在收集有关眼部炎症致病过程的机制，细胞和分子的新信息。收集的信息将用于开发新方法，以治疗这组眼科疾病。 2012财年的研究主要集中在三个问题上：（i）通过抗CD3/CD28抗体的常用激活产生的T-烷（Th）细胞系之间的差异分析，或通过抗原呈递细胞（APC）提出的特定抗原（APC）对生理激活模式的分析； （ii）以针对眼抗原的特异性检查自然和诱导T调节细胞（分别为NTREG和ITREG）的特征； （iii）进一步解剖DAP12对EAU和相关免疫反应的发展的双重双重作用。 概括 该项目针对学习炎症性眼部疾病的致病过程，主要集中在2012财年，主要是在这些过程中涉及的T细胞种群上。具体而言，我们通过关注以下问题来扩展2012财年的研究：（1）通过用抗CD3/CD28抗体激活产生的TH细胞线之间的差异，或者是APC提出的特定抗原； （2）收集有关Treg细胞不同亚群的信息； （3）进一步剖析DAP12对免疫致病反应的令人惊讶的双重作用。 1）不同机制产生的谱系之间的差异。 Th的谱系是通过通过谱系特异性细胞因子同时激活中含中含有CD4细胞与极化产生的。假定在体内，通过中含CD4细胞与其特异性AG的相互作用（由APC提出）提供了激活。由于在制备WT动物的CD4细胞中，仅存在具有特异性的CD4细胞的微小比例，因此，Ag和APC的激活通常是通过将CD4细胞暴露于针对两个细胞分子中的抗体来代替，这些细胞分子参与了生理激活的生理过程，CD3和CD28。 TCR转基因（TG）小鼠的可用性，其中大多数T细胞识别相同的抗原，但是，通过APC提出的特定抗原或抗CD3/CD28抗体，通过激活中含CD4细胞来生成极化Th细胞的线条。这些转基因小鼠使得可以在两种激活模式中的任何一种产生的极化细胞线之间进行比较。 我们比较了由一种激活模式之一产生的Th1，Th9和Th17的谱系，发现在以下生物学活动中，这三种表型的两个谱系之间的两个谱系之间存在显着差异：在受体脾脏中迁移并增殖的能力，重要的是，在接受者中诱导了对受体小鼠眼睛的抗症状，以表达目标antigient antigient antigient antigie antigient antigie antigigient antigigient antigigiend antigigige antigigemen；某些趋化因子，趋化因子受体和主要表面抗原的表达谱；而且，尤其是通过微阵列分析监测的基因表达模式。因此，我们的发现强调了仅用抗CD3/CD28 ABS激活CD4细胞而产生的TH谱系的生物学观察结果的局限性。 2）不同模式产生的T调节（Treg）种群之间的差异。 抑制免疫反应对自我抗原的特定Treg具有用于特异性抑制致病性免疫反应的潜力，并且是强化研究工作的重点。 Treg的诱导要求在极化过程中激活细胞，以获得免疫抑制能力。大多数研究中使用的激活过程是通过针对CD3/CD28的抗体，但是其他研究通过APC提出的抗原采用了特定的激活。据我们所知，没有任何出版物比较了这两个程序产生的Treg细胞。使用我们的TCR转基因小鼠系统（见上文），可以进行这种比较。通常，抗原和APC诱导的Treg细胞比用抗体激活诱导的细胞更具抑制性。在获得极性或极化Th1，Th9或Th17细胞的培养过程中，对Treg的两个谱系的抑制作用进行了测试。在所有系统中，TH17均表现出最高水平的敏感性，而Th1细胞是最具耐药性，TH9细胞的敏感性中间。 3）解决DAP-12对免疫发作反应的神秘双重影响。 DAP-12（12KDA的DNAX激活蛋白）通过传输多种激活信号在免疫反应中起着重要作用。 DAP-12在不同研究中增强或抑制实验性自身免疫性疾病的能力引起了DAP-12在致病自身免疫性病因中的作用。在本研究中，我们比较了DAP-12无效小鼠和野生型（WT）对照，以使其对EAU诱导和免疫反应性的敏感性。 在两个动物设施中饲养小鼠，具有低（a）或高（b）标准。尽管两种设施的WT控制小鼠的疾病发展都是可比的，但在其眼部炎症的严重程度以及其淋巴细胞产生的增殖和细胞因子的水平上，设施A的DAP-12无效小鼠均优于其WT对照。相反，通过两个参数，设施B的DAP-12无效小鼠均低于其对照。 因此，我们的数据为DAP-12：该分子中的缺乏症的作用的争议提供了可能的解决，可以根据环境促进或抑制致病反应。此外，我们建议，两个动物设施的环境之间的差异通过修饰小鼠的微生物菌群，从而影响免疫反应性，从而影响免疫能力。