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

描述(申请人提供)：在过去的几年里，形成T细胞免疫学基础的TH1/TH2范例从TH17细胞的发现开始迅速扩展，TH17细胞是CD4+T记忆细胞的一个子集，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。干燥综合征是一种以外分泌腺功能障碍为特征的自身免疫性疾病，主要累及唾液腺和泪腺。近年来，NOD和NOD来源的小鼠表现出一种与SjS非常相似的疾病，包括伴随唾液和泪腺内白细胞渗入的液体分泌丧失。这种自身免疫性外分泌病分为两个主要阶段：第一，外分泌腺内发生许多独立于自身免疫攻击的病理生理变化；第二，进行性慢性自身免疫反应，导致腺泡细胞质量丧失和外分泌功能下降。NOD-Aec1Aec2小鼠(以及Sjs患者的唾液腺活检组织)的颌下腺免疫组织化学染色显示，淋巴细胞内的IL-17和IL-23都呈强阳性染色，上皮组织也有弥漫的稀疏染色。IL-17和IL-23在C57BL/6小鼠颌下腺中的表达与TH17细胞主控基因ROR3t的表达相关。与此同时，最近的研究表明，下调TH17活性的细胞因子IL-27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小鼠(和SjS患者)的外分泌腺中表达上调，而且可能与该病的临床表现有关。在这项拨款申请中，我们建议研究TH17/IL-17/IL-23-IL-27相互作用系统在SjS的发展和发病中的重要性。为了实现这一目标，提出了两个目标：(I)定义和表征在SjS样疾病发展过程中渗透到唾液腺的IL-23分泌和CD4+TH17记忆T细胞群，以及(Ii)在SjS的C57BL/6.NOD-Aec1Aec2小鼠模型中，确定IL-27对CD4+TH17记忆T细胞群的潜在调节作用，以防止SjS的发展。这项研究的结果有望为深入了解LF相关的TH17/IL-23系统及其调节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(S)提供帮助。识别SjS发生和发展的分子和细胞机制，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，应该指向允许免疫治疗发展的各种新的和新的途径。如果目前的研究结果提供证据表明TH17细胞在SjS公共卫生相关性的发展和/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：几十年来形成T细胞免疫学基础的Th1/TH2范式受到了TH17细胞的发现的挑战，TH17细胞是CD4+T记忆细胞的一个子集，其特征是独特地分泌IL-17家族细胞因子，显然参与了自身免疫。在这项赠款申请中，我们建议利用基因疗法的方法来研究TH17/IL-17/IL-23-IL-27交互系统在干燥综合征(Sjvgren‘s syndrome，SjS)的发生和发展中的可能作用。Sjvgren综合征是一种导致口干和眼睛干燥的自身免疫性疾病。明确SjS发生和发展的分子和细胞机制，特别是TH17/IL-17/IL-23/IL-27调节的生物学变化和与分泌功能障碍相关的分子和细胞机制，应该指向各种新的和根本性的靶点，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.