Fas Ligand Regulation of Ocular Innate Immunity

说明(出自申请者S摘要)：本文拟研究内容如下： (I)确定膜FasL(MFasL)和可溶性FasL(SFasL)在 中性粒细胞介导的眼内炎症，最重要的是，(Ii) 确定sFasL是否可用于下调中性粒细胞介导的 炎症和预防中性粒细胞介导的眼部损伤。初步研究 证明在免疫特权眼内，mFasL激活中性粒细胞 并启动先天免疫，而sFasL抑制中性粒细胞活化 阻止先天免疫力。不过，最令人信服的是sFasL能够 当sFasL和sFasL同时存在时阻断mFasL的促炎作用 MFasL在眼内的表达相同。拟议的研究将利用 一种新的直接在角膜内表达DNA的方法来表征 体内，不同形式的FasL诱导的炎症反应 在角膜基质中表达。MFasL和sFasL都有能力 直接激活的中性粒细胞将在体外使用基质细胞进行评估 分别用不同的FasL cDNA进行转染。最后一组实验将 在体内确定sFasL是否能阻断中性粒细胞介导的炎症 角膜，并在体外阐明sFasL阻断的机制 MFasL可诱导中性粒细胞活化。我们预期这些研究将会 产生关于FasL如何调节眼睛内炎症的新信息，以及 可能导致使用可溶性形式的新疗法的开发 FasL预防中性粒细胞介导的炎症继发眼损伤。

DESCRIPTION (From the Applicant?s Abstract): The studies proposed herein will: (i) determine the roles of membrane FasL (mFasL) and soluble FasL(sFasL) in neutrophil-mediated inflammation within the eye and most importantly, (ii) determine whether sFasL can be used to downregulate neutrophil-mediated inflammation and prevent neutrophil-mediated ocular damage. Preliminary studies demonstrate that within the immune privileged eye, mFasL activates neutrophils and initiates innate immunity, while sFasL inhibits neutrophil activation ad prevents innate immunity. Most compelling though, was the ability of sFasL to block the pro-inflammatory effects of mFasL when both sFasL when both sFasL and mFasL were equally expressed within the eye. The proposed studies will utilize a novel method of expressing DNA directly within the cornea to characterize, in vivo, the inflammatory response induced by the different forms of FasL expressed within the corneal stroma. The ability of both mFasL and sFasL to directly activate neutrophils will be assessed in vitro using stromal cells transfected with the different FasL cDNAs. The final set of experiments will determine, in vivo, whether sFasL blocks neutrophil mediated inflammation in the cornea, and to elucidate, in vitro, the mechanism by which sFasL blocks mFasL induced neutrophil activation. We anticipate that these studies will yield new information as to how FasL regulates inflammation within the eye, and possibly lead to the development of new treatments using the soluble form of FasL to prevent ocular damage secondary to neutrophil mediated inflammation.