2018-09-012024-05-16https://scholars.lib.ntu.edu.tw/handle/123456789/669768摘要：肝臟是一特別的免疫器官，肝臟內調控型T細胞 (Tregs) 會抑制免疫作用細胞而維持對大量進入肝臟的無害的食物和常在菌抗原的耐受性反應。Tregs根據其表型及抑制機轉的不同分成很多種，例如Foxp3 Tregs及Tr1。原發性膽汁性膽管炎(Primary Biliary Cholangitis; PBC)是一種進行性自體免疫性肝臟疾病，其特徵為肝門脈區內小膽管被浸潤到肝臟的作用型T細胞破壞。而PBC發生過程中Tregs功能及角色則尚不清楚。此外，若能引起肝臟Tregs的功能，對於恢復肝臟的耐受性及控制 PBC是一極好的治療策略。在此研究計劃中，我們將利用我們研究室先前建立的2-OA-OVA致敏的小鼠模式探討在 PBC中肝臟Tregs的角色以及細胞激素IL-2及IL-30在 PBC 的治療效果，進一步探討這治療效果是否與促進Tregs有關。三個子目標如下:子目標一：探討在 PBC疾病進程中肝臟Tregs的角色1.分析PBC疾病不同時期肝臟Tregs的特徵及動態變化2.探討剔除Tregs對 PBC發病及肝臟免疫反應的影響子目標二：使用重組腺相關病毒攜帶IL-2基因(AAV-IL-2) 探討IL-2對PBC的作用1.重組腺相關病毒攜帶IL-2基因在小鼠體內能否增加及活化肝臟Tregs。2.給予AAV-IL-2能否降低 PBC的發生3.給予AAV-IL-2能否治療已發生的 PBC 子目標三：使用重組腺相關病毒攜帶IL-30基因(AAV-IL-30) 探討IL-30對PBC的作用1.探討IL-30在 PBC中的治療效果2.探討IL-30是否藉由引起IL-10或Tr1而降低 PBC這研究將釐清在自體免疫膽管炎不同時期的肝臟內Foxp3 Tregs和Tr1的角色，以及IL-2和IL-30的免疫調節功能及機轉。此研究結果不僅可以了解肝臟發炎時肝臟內effector T cells/Tregs的角色及免疫機轉，亦提供肝臟自體免疫疾病的治療策略。<br> Abstract: Liver is a special immune organ with delicate and finely tuned balance between tolerance to harmless dietary and commensal antigens and immunity to pathogens. Liver contains different subsets of effector lymphocytes that are kept in check by a subpopulation of T cells known as regulatory T cells (Tregs). Several Treg subsets, such as Foxp3 Tregs and Tr1, can be identified by their phenotype and suppressive mechanisms. Primary biliary cholangitis (PBC) is a progressive autoimmune liver disease mediated by immune-mediated destruction of intrahepatic small bile ducts. Liver-infiltrating effector T cells are the major player for the destruction of biliary cells. However, the role of various hepatic Treg subsets during the development of PBC is still unclear. In addition, the induction of hepatic Tregs is an attractive therapeutic strategy for restoring self-tolerance and controlling PBC.In the current application, we will investigate the role of hepatic Treg subsets and the therapeutic effects of cytokines IL-2 and IL-30 on PBC using our previous established murine model of 2-OA-OVA immunization. In addition, we will elucidate whether Treg subsets are involved in the IL-2 and/or IL-30 therapy. Specific aim 1. To investigate the role of the hepatic Tregs in the progression of PBC by 2-OA-OVA immunized mice. Aim 1.1 To evaluate the characteristics and dynamic changes of hepatic Treg subsets during the development of PBC in 2-OA-OVA immunized mice.Aim 1.2 To define the overall effect of Foxp3 Tregs on the adaptive and innate immune responses of PBC by Foxp3 Tregs depleted mice and determine how they may influence the outcome of PBC.Specific aim 2. To investigate the effect of IL-2 on PBC in 2-OA-OVA immunized mice using recombinant adeno associated virus (AAV) vector encoding IL-2Aim 2.1 To determine whether IL-2 administration by AAV delivery expands and activates hepatic Tregs in vivo.Aim 2.2 To investigate whether expansion of Tregs by AAV-IL-2 administration impairs PBC induction.Aim 2.3 To investigate whether expansion of Tregs by AAV-IL-2 administration impairs established PBC. Specific aim 3. To investigate the effect of IL-30 on PBC in 2-OA-OVA immunized mice using recombinant adeno associated virus (AAV) vector encoding IL-30Aim 3.1 To define the therapeutic effects of IL-30 in PBC.Aim 3.2 To determine whether IL-30 induces IL-10 or Tr1 in reducing PBC.This study will clarify the role of hepatic Foxp3 Tregs and Tr1 in different phases of autoimmune cholangitis and the immunosuppressive effects and mechanism of IL-2 and IL-30. The results of this study will be of importance for not only dissecting the role of Tregs in the balance between immunity and tolerance in liver inflammation but also providing immunotherapy strategy for liver autoimmune diseases.肝臟自體免疫疾病調控型T細胞重組腺相關病毒小鼠模式介白素2介白素30liver autoimmune diseaseregulatory T cellsrecombinant adeno associated virusmouse modelIL-2IL-30