2013-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649034摘要：有許多已知的自體免疫疾病都會侵犯到人體的血管，而出現血管炎的情形。而以前針對這些血管炎在臨床上都沒有較好的評估方法，所以在我們的計畫中將研究與血管炎相關的抗體和血中的內皮細胞數目。我們將在此一計畫中建立新的方法來追蹤患者血中與血管炎相關的自體抗體；同時我們也將建立利用微流技術的方法來測定血中的游離內皮細胞數目，並進一步來了解是否在這些患者身上抗內皮細胞抗體濃度和血中游離內皮細胞數目是否可以與患者血管炎的程度相對照。此一跨領域的研究計畫主要是結合應用所老師的研究專長，可以利用細胞大小、標定後磁力和流速等因子來偵測和分離細胞。而在臨床應用上，我們計畫在臨床上以一個孩童期最常見的血管炎，過敏性紫斑(Henoch-Scholein purpura)再加上最常見的自體免疫疾病如全身性紅斑狼瘡為研究的模式，以了解是否可以流體動力學上的技術來研究這些患者身上的內皮細胞，利用流體動力學的技術建立起全新的診斷方法。在過敏性紫斑的患童會出現抗內皮細胞的抗體，而且與這些小朋友的臨床症狀來加以對照，以了解這些自體抗體與血中循環的內皮細胞數目是否有任何相關性。此外，我們將利用這些新建立的技術和方法來研究另外一個最典型的自體免疫疾病，全身性紅斑狼瘡，的患者身上的自體抗體和循環中的內皮細胞數目，以了解是否與這些全身性紅斑狼瘡患者的疾病活性有關。第一年： 我們將利用建立評估方法來測定自體免疫疾病，尤其是先研究在孩童最常出現看到的過敏性紫斑(Henoch-Scholein purpura)。過敏性紫斑的主要表現是抗體導致血管炎，所以會出現出血點、紫斑、腹痛、關節痛和腎炎，但是一直沒有一個較好的檢驗方法可以作為過敏性紫斑的臨床指標。我們將在第一年的計畫中建立評估方法可以應用到自體免疫疾病相關的血管炎研究上，作為重要的臨床指標。第二年：我們將利用微流技術來建立方法可以測定自體免疫疾病患者血中的內皮細胞數目，所以在第二年的計畫中我們將利用此一技術來建立方法測定內皮細胞的數目，以了解是否與自體疾病的血管炎發炎有關。此種微流技術未來可以測定血中非常少數的細胞，所以我將利用這種微流技術建立的方法來分析患者血中游離內皮細胞的數目，與體內血管發炎的程度做個對照。第三年：我們將利用建立好的方法也應用在其他自體免疫疾病，如全身性紅斑狼瘡，以了解這些抗內皮細胞抗體和血中游離的內皮細胞在這些可能具有血管炎的自體免疫疾病患者身上所具有的意義。如果我們可以在這些會造成血管炎的自體疾病可以測到這些相關的自體抗體，和能夠再測定出游離的內皮細胞，與患者的血管炎嚴重性來加以評估對照，相信一定可以讓我們對這些疾病的機轉和未來的治療有更清楚的了解。在此一計畫中，我們將建立新的方法來測定與血管炎相關的自體抗體，尤其是發展出一個新的方法可以來測定微量的游離內皮細胞。我們將利用這些新建立好的方法來評估是否能夠找到一個較好的方法來評估發生在自體免疫疾病的血管炎，如果能夠順利在過敏性紫斑和全身性紅斑狼瘡這些會出現血管炎的患者可以有效地利用這些新研發出來的方法來進行評估，未來也可能有機會應用到其他如心血管和腫瘤疾病的應用上。<br> Abstract: In this study, we like to investigate the immunological changes in children with autoimmunevasculitis with determination of circulating endothelial cells with the techniques ofmicro-fluidics. We like to include children with Henoch-Scholien purpura and systemic lupuserthymatosus in the study. Henoch-Scholein purpura has been found to the most frequentvasculitis in childhood. Most of the patients experienced upper respiratory tract or othergastrointestinal tract infection one or two weeks before the manifestation of the symptomssuch as purpura, abdominal pain and arthralgia. Among the laboratory parameters, IgAanti-endothelial cell antibody has been found to play a critical role in the pathogenicmechanism of Henoch-Scholein purpura. With the methods and techniques of micro-fluidics,we might be able to investigate the percentage of circulating endothelial cells in the patients.Further, we could be also able to analyze the frequency of endothelial cells and correlate withthe antibody level and disease activities. We believe the collaboration between our team andexperts of micro-fluidics could shed light on exploring the novel diagnostic tool forautoimmune vasculitis such as Henoch-Scholein purpura and systemic lupus erythematosus.First year: We like to study the level of anti-endothelial autoantibodies in the patients withHenoch-Scholein purpura. Henoch-Scholein purpura is the most common vasculitis inchildhood. Especially, we like to focus on the assessment of anti-endothelial cells antibodiesin patients with Henoch-Scholein purpura and systemic lupus erythematosus and analyze thecorrelation of clinical severity with the level of anti-endothelial cell antibody.Second year: We aim to establish the novel method to determine the level of circulatingendothelial cells in the children with autoimmune vasculitis such as Henoch-Scholein purpuraand systemic lupus erythematosus. Flurocytometric assay is the conventional method todetermine the number of circulating endothelial cells, however, the sensitivity of the methodis still not good enough to assess the small number of circulating endothelial cells. In thesecond year, we like to establish the microfluidic technique for the determination ofcirculating endothelial cells. Further, we like to assay the levels of anti-endothelial cellsantibody and compared with the number of circulating endothelial cells. We like to analyze ifthe levels of circulating endothelial cells are correlated with the disease activity ofHenoch-Scholein purpura.Third year: We like to investigate the levels of anti-endothelial cells antibodies and circultingendothelial cells in the children with systemic lupus erythematosus. In addition, we also liketo study the condition of endothelial cells to see if these circulating endothelial cells areaffected by anti-endothelial cells. With the microfludic method, we will be able to analyze theapoptotic signal pathways of endothelial cells in the patients with autoimmune vasculitis.In this project, we plan to develop the methods to detect anti-endothelial antibodies and alsoclinical severity for monitoring the immune-mediated vasculitis. Further, we also like todesign and develop the novel method with microfluidic techniques to detect the number ofcirculating endothelial cells. We will correlate the levels of autoantibodies and circulatingendothelial cells with the disease activity. We believe the novel assays derived from thisproject will shed light on further understanding the pathogenic mechanisms of the vasculitisoccurred in autoimmune diseases.