Gene therapy targeting the Th17 / IL-27 system in autoimmune exocrinopathy

描述（由申请人提供）：在过去的几年中，TH1/TH2范式形成了T细胞免疫学的基础，从TH17细胞的发现开始迅速扩展，TH17细胞是CD4+ T记忆细胞的一个亚群，其特征是具有分泌IL-17家族细胞因子的独特能力。最重要的是，TH17细胞似乎与先天免疫和自身免疫密切相关。干燥综合征（Sjvgren's syndrome, SjS）是一种主要影响唾液腺和泪腺的自身免疫性疾病，以外分泌腺功能障碍为特征。近年来，NOD和NOD衍生小鼠表现出一种与SjS非常相似的疾病，包括唾液和泪腺内出现白细胞浸润的液体分泌物丧失。这种自身免疫性外分泌病可分为两个主要阶段：第一阶段，许多病理生理变化发生在外分泌腺内，独立于任何自身免疫攻击；第二阶段，进行性慢性自身免疫反应导致腺泡细胞团丧失和外分泌功能下降。C57BL/6颌下腺免疫组化染色。NOD-Aec1Aec2小鼠（以及SjS患者的唾液腺活检）现在显示淋巴细胞灶内IL-17和IL-23的强烈阳性染色，以及上皮组织的弥漫性稀疏染色。IL-17和IL-23在C57BL/6颌下腺组织中的表达。NOD-Aec1Aec2小鼠与TH17细胞主调控基因ROR3t的表达相关。与此同时，最近的研究表明，IL-27，一种下调TH17活性的细胞因子，在外分泌腺中的表达水平非常低。这些结果表明，TH17/IL-17/ IL-23系统不仅在C57BL/6的外分泌腺中上调。NOD-Aec1Aec2小鼠（和SjS患者）在发病时，但可能有助于疾病的临床表现。在本次资助申请中，我们建议研究TH17/IL-17/ IL-23 - IL-27相互作用系统在SjS发展和发病中的重要性。为了实现这一目标，我们提出了两个目标：(i)定义和表征SjS样疾病发展过程中浸润涎腺的IL-23分泌和CD4+ TH17记忆T细胞群；（ii）在C57BL/6中确定IL-27对CD4+ TH17记忆T细胞群的可能调控潜力，从而使用基因治疗方法预防SjS的发展。NOD-Aec1Aec2小鼠SjS模型。本研究的结果有望深入了解与lf相关的TH17 / IL-23系统及其调节的IL-27系统在导致唾液腺功能障碍的自身免疫反应的发展和可持续性中的相互关系。识别参与SjS发展和发病的分子和细胞机制，特别是在分子水平上关注与分泌功能障碍相关的生物学变化，应该指出各种新的和新颖的途径，允许开发免疫治疗。如果目前的研究结果提供证据表明TH17细胞在SjS的发展和/或发病中发挥积极作用，那么通过IL-27调节TH17/IL-17/ IL-23系统的基因治疗方法将是一种高度可行的方法。TH1/TH2范式几十年来一直是T细胞免疫学的基础，但随着TH17细胞的发现，TH1/TH2范式受到了挑战。TH17细胞是CD4+ T记忆细胞的一个亚群，其特点是具有分泌IL-17家族细胞因子的独特能力，并明显参与自身免疫。在本次拨款申请中，我们建议利用基因治疗方法研究TH17/IL-17/ IL-23 - IL-27相互作用系统在干燥综合征（Sjvgren's syndrome, SjS）的发展和发病中的可能作用，干燥综合征是一种导致口干和干眼综合征的自身免疫性疾病。确定SjS发生和发展的分子和细胞机制，特别是关注TH17/IL-17/ IL-23 / IL-27调控的生物学变化，以及与分泌功能障碍相关的生物学变化，应该指向各种新的和激进的靶点，从而允许开发免疫疗法来治疗SjS。

DESCRIPTION (provided by applicant): Over the past several years, the TH1/TH2 paradigm forming the basis of T cell immunology has expanded rapidly from the discovery of TH17 cells, a subset of CD4+ T memory cells characterized by their unique ability to secrete IL-17 family cytokines. Most importantly, TH17 cells appear to be intimately involved in innate immunity and autoimmunity. Sjvgren's syndrome (SjS) is an autoimmune disease affecting primarily of the salivary and lacrimal glands characterized by exocrine gland dysfunction. In recent years, NOD and NOD-derived mice have been shown to exhibit a disease that closely parallels SjS, including the loss of fluid secretions concomitant with the appearance of leukocyte infiltrates within the salivary and lacrimal glands. This autoimmune exocrinopathy has been separated into two major phases: first, numerous pathophysiologic changes occur within the exocrine glands independent of any autoimmune attack, and second, a progressive chronic autoimmune response resulting in loss of acinar cell mass and decline in exocrine function. Immunohistochemical staining of submandibular glands from C57BL/6.NOD-Aec1Aec2 mice (as well as salivary gland biopsies from SjS patients) has now shown strong positive staining for both IL-17 and IL-23 within the lymphocytic foci, plus a diffuse, sparsely staining of epithelial tissues. Temporal expressions of IL-17 and IL-23 in submandibular glands of C57BL/6.NOD-Aec1Aec2 mice correlated with expression of ROR3t, the TH17 cell master control gene. At the same time, more recent work has shown that IL-27, the cytokine that down-regulates TH17 activity, is expressed at very low levels in the exocrine glands. These results suggest that the TH17/IL-17/ IL-23 system is not only up-regulated in the exocrine glands of C57BL/6.NOD-Aec1Aec2 mice (and SjS patients) at time of disease, but may contribute to the clinical manifestations of the disease. In this grant application, we propose to study the importance of this TH17/IL-17/ IL-23 - IL-27 interactive system in the development and onset of SjS. To achieve this goal two aims are advanced: (i) define and characterize the IL-23 secreting and CD4+ TH17 memory T cell populations infiltrating the salivary glands during development of SjS- like disease, and (ii) determine a possible regulatory potential of IL-27 on the CD4+ TH17 memory T cell populations for preventing development of SjS using a gene therapy approach in the C57BL/6.NOD-Aec1Aec2 mouse model of SjS. Results from this study are expected to provide insight into the inter-relationship(s) between the LF-associated TH17 / IL-23 system and its regulatory IL-27 system in the development and sustainability of the autoimmune response leading to salivary gland dysfunction. Identification of molecular and cellular mechanisms involved in the development and onset of SjS, focusing specifically on biological changes associated with secretory dysfunction at a molecular level, should point to a variety of new and novel pathways permitting development of immunotherapy. A gene therapy approach directed towards regulating the TH17/IL-17/ IL-23 system through IL-27 would be a highly feasible method if results of the current study provide evidence that TH17 cells play an active role in the development and/or onset of SjS PUBLIC HEALTH RELEVANCE: TH1/TH2 paradigm, forming the basis of T cell immunology for decades, has been challenged by the discovery of TH17 cells, a subset of CD4+ T memory cells characterized by their unique ability to secrete IL-17 family cytokines and apparently involved in autoimmunity. In this grant application, we propose to utilize a gene therapy approach to study the possible role of the TH17/IL-17/ IL-23 - IL-27 interactive system in the development and onset of Sjvgren's syndrome (SjS), an autoimmune disease leading to dry mouth and dry eye syndromes. Identification of molecular and cellular mechanisms involved in the development and onset of SjS, focusing specifically on biological changes regulated by TH17/IL-17/ IL-23 / IL-27 and associated with secretory dysfunction, should point to a variety of new and radical targets permitting development of immunotherapy to treat SjS.