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，形成T细胞免疫学基础的TH 1/TH 2范例已经从TH 17细胞的发现中迅速扩展，TH 17细胞是以其分泌IL-17家族细胞因子的独特能力为特征的CD 4 + T记忆细胞的子集。最重要的是，TH 17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衍生的小鼠已被证明表现出与SjS密切平行的疾病，包括伴随唾液腺和泪腺内出现白细胞浸润的液体分泌物的损失。这种自身免疫性外分泌病分为两个主要阶段：第一，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，进行性慢性自身免疫反应导致腺泡细胞质量损失和外分泌功能下降。来自C57 BL/6. NOD-Aec 1Aec 2小鼠的下颌下腺（以及来自SjS患者的唾液腺活检）的免疫组织化学染色现在显示淋巴细胞病灶内的IL-17和IL-23的强阳性染色，加上上皮组织的弥漫性稀疏染色。IL-17和IL-23在C57 BL/6. NOD-Aec 1Aec 2小鼠下颌下腺中的时间表达与TH 17细胞主控基因ROR 3 t的表达相关。与此同时，最近的研究表明，IL-27，下调TH 17活性的细胞因子，在外分泌腺中以非常低的水平表达。这些结果表明，TH 17/IL-17/ IL-23系统不仅在C57 BL/6. NOD-Aec 1Aec 2小鼠（和SjS患者）的外分泌腺中在疾病时上调，而且可能有助于疾病的临床表现。在这项资助申请中，我们建议研究TH 17/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，提出了两个目标：（i）定义和表征在SjS样疾病发展过程中浸润唾液腺的IL-23分泌和CD 4 + TH 17记忆T细胞群，和（ii）确定IL-27对CD 4 + TH 17记忆T细胞群的可能调节潜力，用于在C57 BL/6.NOD中使用基因治疗方法预防SjS的发展。Aec 1Aec 2小鼠SjS.本研究的结果预计将深入了解LF相关TH 17/ IL-23系统及其调节性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。鉴定参与SjS的发展和发病的分子和细胞机制，特别是在分子水平上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，允许免疫治疗的发展。如果当前研究的结果提供证据表明TH 17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，则通过IL-27调节TH 17/IL-17/ IL-23系统的基因治疗方法将是一种高度可行的方法：数十年来形成T细胞免疫学基础的TH 1/TH 2范式受到了TH 17细胞的发现的挑战，CD 4 + T记忆细胞的一个亚群，其特征在于其独特的分泌IL-17家族细胞因子的能力，并明显参与自身免疫。在这项资助申请中，我们建议利用基因治疗方法来研究TH 17/IL-17/ IL-23 - IL-27相互作用系统在干燥综合征（SjS）（一种导致口干和干眼综合征的自身免疫性疾病）的发展和发病中的可能作用。鉴定参与SjS的发展和发病的分子和细胞机制，特别关注由TH 17/IL-17/ IL-23 / IL-27调节的生物学变化，并与分泌功能障碍相关，应该指出各种新的和激进的目标，允许发展免疫疗法来治疗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.