2010-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/657902摘要：異位性皮膚炎屬於慢性的發炎性疾病‚它的主要特色之一就是反覆的發作‚造成臨床控制的一大難題;臨床上廣泛使用的抗發炎製劑僅提供症狀的緩解,但是卻帶來相當大的副作用。我們因此有必要研究其致病機轉‚特別是針對過敏原被呈現以及抗原呈現細胞調控免疫反應等來研發專一性的治療策略。已知過敏原的呈現及免疫的調控和蘭格罕細胞(Langerhans cell)上的分子-蘭格素Langerin (CD207)有關‚目前有證據顯示某些抗原的確會經由蘭格素進行吸收(uptake),但是其角色及功能上的意義尚未清楚。我們過去採用多重光子顯微術(Cr:forsterite fslaser-based multimodality nonlinear microscopy)來從事相關的研究‚已經證明了此一技術應用在異位性皮膚炎的可行性;我們希望進一步發展活體成像技術(in vivo imaging),以此取代目前許多研究異位性皮膚炎的in vitro 方法,並且利用單株抗體(monoclonal antiby)來標定(target)、追蹤(trace) 以及阻斷(block)或修飾(modify)蘭格素相關的抗原呈現以及免疫調控的作用。我們假設OVA 抗原經由蘭格素進行吸收終究導致Th2 T 淋巴球的活化,進而引起異位性皮膚炎的慢性發炎反應。因此,我們採用in vivo imaging 與immunohistochemistry,一方面決定蘭格素的表現量,一方面也以co-localization study 來決定抗原經由蘭格素進行吸收。最後我們預期可以藉由單株抗體去阻斷(block) 蘭格素相關的抗原呈現以及免疫調控的作用,我們將比較阻斷蘭格素前後的異位性皮膚炎的皮膚組織學以及免疫血清學的變化,以此驗證我們的假說。我們認為多重光子顯微術是一種安全的活體成像(in vivo imaging)技術‚可以應用於臨床及基礎研究,也可以促進我們去實際瞭解活體中異位性皮膚炎的詳細致病機轉, 研發真正針對特定分子細胞的疾病治療策略,有助於找出以免疫療法為理論基礎的異位性皮膚炎長期控制之道。<br> Abstract: Atopic dermatitis represents an eczematous disorder of the skin, which affects up to 20%of children and 1~3% of adults. Atopic dermatitis is characterized by chronic eczematouslesions, typically involving the flexural folds which are impaired by recurrent severe flare upsof AD. as most of the treatments of AD are limited to symptomatic therapies such asanti-inflammatory or anti-pruritic therapy only. Obviously, therapeutic approaches whichselectively regulate aberrant pathophysiological mechanisms in AD would be much moreeffective and promising.The most prominent members of dendritic cells are the classical Langerhans cells (LCs),which are characterized by the Birbeck granules, electron microscopically visible as tennisracket-shaped organelles originating from the accumulation of the C-type lectin langerin(CD207). LCs resided in both healthy and inflamed skin and were constantly renewed understeady-state conditions. An important role of LC can be allocated to the initial phase of AD.In this step, primarily T cells of the Th2 type are primed by LCs in vitro, which arecharacterized by the production of IL-4, IL-5, and IL-13, typical for the initial phase of AD.Langerin is a 40-kD protein and belongs to a member of C‑type lectin receptors (CLRs) withmannose-binding specificity. Langerin is a type II transmembrane cell surface receptor onLCs and is restricted to them. It has been suggested that the sensitization process of antigencapture, internalization, processing, migration to lymphoid tissue, and maturation of LCs is aconsequence of langerin function.We aimed to set-up an in vivo imaging method to investigate the antigen uptake ofsensitization process and immunomodulation pathway underlying the pathophysiology of ADwhich was selectively regulated by langerin on LCs. We use Cr:forsterite fs laser-basedmultimodality nonlinear microscopy as our non-invasive tool. We first determine theexpression level of langerin, then tracking the dynamics of LCs and the uptake of OVA usingin vivo imaging techniques. With fluorescence immunohistochemistry and co-localizationstudy, we aimed to validate the uptake of external antigens (OVA) by epidermal Langerhans cells (LCs) were mediated through langerin. We hypothesized that allergen (OVA) uptake bylangerin play a central role in the shaping and further development of allergic sensitization inAD. Therefore, with monoclonal antibodies targeting and blocking langerin, we also wishedto develop a long-term management strategy of AD to eliminating the recurrent-relapsingcourse of AD.異位性皮膚炎atopic dermatitisCD207epicutaneous sensitizationlangerinLangerhans cellmurine modelovalbumintwo-photon fluorescence