2013-01-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649312摘要：過敏疾病在過去幾年來可以說是逐年增加，已經成為所有國民健康上的一個重要問題，所以如何更進一步了解過敏疾病的發生機轉和研發出更有效的治療方法是一個刻不容緩的研究課題。目前已經知道過敏疾病的主要免疫調節是跟第一型與第二型T輔助細胞的調控有關，第二型T輔助細胞所分泌的IL-4和IL-5會分別刺激IgE抗體及嗜伊紅性白血球的產生而導致發炎反應。所以，研究出更有效的治療方法和了解這些治療方法的機轉將有助於我們在未來研發出更有效的治療方法。而在我們之前的研究中發現利用口服方式給予過敏原對降低呼吸道的發炎反應有很重要的影響，因此我們進一步研究黏膜免疫系統中的細胞在口服耐受性中所扮演的角色。此外，我們發現如果利用B細胞與T細胞一道培養後可以有效地誘發出具有調節能力的T細胞。我們在此一計畫中要來進一步研究這些由B細胞誘發出來的調節性T細胞(Treg/B cells)在黏膜免疫中所扮演的角色，同時也要研究這些Treg/B細胞的功能和基因表現，未來如何來誘發這一群細胞能夠應用到免疫疾病的調控上。第一年：我們之前已經證明利用口服餵食過敏原的確可以有效地降低過敏原特異性的抗體，改善呼吸道的發炎反應和減少呼吸道阻力。同時，我們也將進一步研究是否能夠利用B細胞來誘發出具有調節能力的T細胞，再研究這些T細胞的特性和表面標記的表現。此外，我們也將分離不同的抗原呈現細胞與T細胞一道培養，以了解這些調節性T細胞是否也能夠像B細胞一樣誘發具有調節能力的細胞。第二年：在這一年的計畫中，我們將這些Treg/B細胞注射入氣喘的小鼠體內，以了解是否能夠改善氣喘小鼠的呼吸道發炎，尤其是降低過敏原特異性抗體和改善呼吸道發炎反應。此外，我們也將分離出不同活化和分化階段的B細胞與T細胞一道培養，如B-1和B-2細胞，活化和未活化的B細胞，記憶性B細胞，以了解哪些B細胞能夠誘發出具有調節能力的T細胞。第三年：我們將利用基因晶片的方法來研究是否能夠在這些由B細胞誘發的調節性T細胞找到表現較高的基因，尤其是那些傳統認知的調節性T細胞沒有表現的分子。一但找到這些特殊的基因後，我們將分別利用real-time PCR來更進一步加以證實，並利用阻斷性抗體來研究是否能夠抑制其調節性反應。此外，我們將進一步分離出這些特定表面標記的Treg/B細胞應用到氣喘的小鼠動物模式，以了解這些特定的Treg/B細胞是否能夠改善呼吸道的發炎反應。在此一計畫中，我們發現由黏膜系統分離出的B細胞與T細胞一道培養後可以誘發出具有調節能力的T細胞。我們計畫研究這群由B細胞所誘發出的調節性T細胞(Treg/B cells)的特性和基因表現，也要進一步將這些Treg/B細胞轉送入氣喘的小鼠，以了解是否能夠抑制過敏免疫反應。這群由B細胞所誘導出來的調節性T細胞可能在整個免疫反應的回饋性調控上也扮演著一個重要的角色，因此更值得加以研究。我們相信這群新的調節性T細胞如果能夠研究地更清楚，將有助於未來進一步將這些細胞應用到臨床免疫疾病的治療上。<br> Abstract: Asthma is one of the most common chronic airway inflammatory diseases. Allergen specific immunotherapy (SIT) has been used for allergic diseases treatment and while the induction of immunologic tolerance against non-pathogenic antigen is a phenomenon observed along the mucosa of the respiratory, gastrointestinal, and urogenital tract, its mechanism remains unclear. Our study demonstrated that oral tolerance cound be used for the treatment of allergic diseases. Among the mucosal immune cells, B cells are the most cell population in Peyer’s patches. Our preliminary studies demonstrated that T cells cultured with B cells could exert the suppressive effect on immune functions. In this study, we aim to study further on the characteristics and functions of regulatory T cells induced by B cells.First year: We like to study the functions of regulatory T cells induced by B cells (Treg/B cells) on the effector T cells. We like characterize the surface markers and cytokine profile of these Treg/B cells, especially the in vitro suppressive activity. In addition, we also like to compare the ability of different antigen presenting cells and B cells from different subsets of B cells in inducing Treg/B cells.Second year: In this year, we aim to isolate Treg/B cells for the in vivo experiment to test if these Treg/B cells could alleviate the airway inflammation and hyperresponsiveness in animal model of asthma. We aim to study further on the activation status and developmental stages of B cells on the induction of regulatory T cells. We like to isolate the B cells with different activation markers and cultured with T cells to find out the possible factors of B cells can affect the development of Treg/B cells. Third year: This year, we like to study the expression of certain genes in the regulatory T cells induced by B cells. We plan to investigate the particular unique genes expressed by microarray analysis. In addition, we also like to study the role of certain genes with available gene knockout mice. Finally, we like to further study the in vivo functions of certain surface markers positive Treg/B cells with adoptive transfer experiment.Many papers on Foxp-3 expressing regulatory T cells published in the past several years, however, regulatory T cells induced by B cells as we reported has been less studied. This project might shed light on further understanding the functions of Treg/B cells and their potential application in clinical immune diseases.黏膜耐受性氣喘調節性T 細胞Asthmamucosal B cellsregulatory T cells induced by B cells