2011-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/660872摘要：異位性皮膚炎是一種常見的慢性發炎性皮膚疾病，異位性疾病的過敏原皆為環境中普遍存在的蛋白質抗原。蛋白質抗原經皮致敏已知為異位性皮膚炎重要的致敏途徑之ㄧ。我們和其他學者利用小鼠模型的研究，也證實蛋白質抗原經皮致敏引發顯著的Th2 型反應，少量的Th17 和Th1 反應。半抗原經皮致敏引發的過敏性接觸性皮膚炎也是皮膚的重要疾病之一，半抗原的經皮致敏，引發的是Th1和Tc1 為主的反應。目前已知 CD4 T 細胞可以分化為多種亞型，藉由分泌不同的cytokine，而參予不同的免疫反應。調控Th 分化的控制因子中，DC 一直被認為具有關鍵的角色，DC 可以藉由表面的costimulator 的表現和釋出的cytokine，來調控Th 細胞的活化和分化。這些年來，關於DC subset 的研究，有很大的進步，目前已知在spleen 中，除了plasmacytoid DC (pDC)外，有三種conventional DC，分別是CD4+DC，CD8+DC，double negative DC (dnDC)。而在periphery lymph node (LN)內，除了上述的DC subset 外，還有數種skin-derived migratory DC，分別是LC，langerin+ dDC，langerin- dDC。目前已知這些不同的DC subset 可能同時執行overlap 和unique 的功能。BALB/c 易引發Th2 型，而B6 易引發Th1 型反應，因此BALB/c 和B6 小鼠分別對應人類的atopy 和non-atopy。在BALB /c 和B6 小鼠中，各種DC subset的數目和性狀不同，屢被報告。因此，我們決定比較分析 (1) BALB/c 和B6 小鼠 (2)蛋白質抗原和半抗原，在經皮致敏的免疫反應中，各DC subset 的角色及功能。我們將運用各種實驗方法，來達成下列特定目標: 特定目標一: 探討naïveBALB/c 和B6 小鼠的spleen 和regional lymph nodes (LN)內，各種dendritic cellsubset (DC subset)的數目和性狀的異同。特定目標二: 探討BALB/c 和B6 小鼠，分別接受蛋白質抗原或半抗原經皮致敏後，其LN 內各種DC subset 的數目和性狀改變的異同。特定目標三: 探討BALB/c 和B6 小鼠，在蛋白質抗原和半抗原經皮致敏的過程中，被各種DC subset 捕捉並呈獻的異同。我們預期此研究計畫的成果，將非常有助於我們了解 DC 在調控免疫反應Th1/Th2/Th17/iTreg 的角色及機制，而且可以應用於異位性疾病蛋白質過敏原和過敏性接觸性皮膚炎半抗原過敏原經皮致敏的預防，以及應用於異位性疾病蛋白質過敏原疫苗的研發。<br> Abstract: Atopic dermatitis is a common chronic cutaneous inflammatory disease. Theallergens of atopic disease are proteins universally present in environment.Epicutaneous sensitization with proteins allergen has been proven to be one of themost important routes for sensitization. We and other researchers have established amurine patch model and demonstrated that epicutaneous sensitization with proteinantigen induces a predominant Th2, a modest Th17 and a weak Th1 responses.Hapten-induced allergic contact dermatitis is also a common cutaneous inflammatorydisease. Epicutaneous sensitization with hapten induces Th1 and Tc1 responses.There are several T helper (Th) cell subsets with different functions. Dendriticcells play a critical role in Th differentiation by means of their costimulatorexpression and cytokine release.There are some DC subsets too. At present，four DCsubsets are identified in spleen. Peripheral lymph nodes have several migratory DCsubsets in addition. Each DC subset may perform overlapping and unique function.For example, CD8+ DCs are superior in cross-presentation to CD8 T cells.Th2-prone BALB/c and Th1-prone B6 mice are correspondents of human atopyand non-atopy, respectively. It has been repeatedly reported that the numbers andcharacters of DC subsets are different between BALB/c and B6 mice. Thus, wesought to investigate the roles of DC subset in epicutaneous sensitization with proteinantigen and hapten in BALB/c and B6 mice. We will use the following technique,including flow cytometic analysis and sorting, real- time PCR, ELISA and so on, toachieve three specific aims.1. Explore the differences of numbers and characters of DC subsets among naïveBALB/c and B6 mice.2. Explore the difference of number and character of DC subset among BALB/cand B6 mice after epicutaneous sensitization with protein antigen or hapten.3. Explore the difference of antigen distribution in DC subsets among BALB/c andB6 mice after epicutaneous sensitization with protein antigen or hapten.We believe that the results obtained from this project will have a high impack onour understanding of mechanism controlling Th1/Th2/Th17/iTreg differentiation. Inaddition, the results will provide useful information for prevention of epicutaneoussensitization with allergens as well as development of vaccine for atopic diseases.