CUTANEOUS BIOLOGY OF NITRIC OXIDE

一氧化氮(NO)被认为是一种重要的信使和效应器 皮肤生理和动态平衡中的分子以及NO水平的变化 在皮肤上引起疾病。该提案的目标是定义角色 皮肤中的一氧化氮，重点是确定其功能效应 这种气体。 人类，而不是小鼠，角质形成细胞已被证明具有 表达诱导型一氧化氮合酶(INOS)和 生产编号：老鼠和人之间的这种关键差异可能解释了为什么 小鼠不会出现类似于在小鼠身上发现的皮疹 人类。将一氧化氮合酶定位于模拟皮肤的转基因小鼠 这里显示的是在人类条件下发现的发展表型。在传统上 小鼠，iNOS在郎格汉斯细胞(LC)中表达。NO也是由 类LC细胞系XS-52。LC中产生的NO可能会影响LC的功能， 由于NO可以自由扩散到细胞膜上，因此它有能力 影响表皮中的相邻细胞。一氧化氮对黑素细胞的毒性作用 角质形成细胞提示NO在许多情况下可能是效应分子。 皮肤状况，包括炎症后色素沉着， 白癜风、移植物抗宿主病(GVH)、硬皮病和通常致命的 过程中，毒性表皮坏死松解(十)。 在拟议的研究中，将对纯化的LC进行生产评估 用RT-PCR检测诱导型一氧化氮合酶的表达，用Griess反应和 放射性L-精氨酸转化为瓜氨酸。的影响 选定的细胞因子在一氧化氮合酶的调节中将被检测。其影响 NO对XS-52细胞本身的影响将被研究。NO的作用机制-- 黑素细胞和角质形成细胞的诱导杀伤将在联合 XS-52细胞培养实验和无供体孵育实验。 将一氧化氮合酶表达定位于表皮的转基因小鼠 产生了，表现为白发和组织病理学， 硬皮病。这些动物将被进一步描述。小鼠表达 在可诱导启动子的控制下将产生NO。这个 转基因小鼠将作为色素沉着障碍的模型进行测试， 抗原提呈、硬皮病、TEN和GVH。这些结果具有 可能导致新的治疗策略的治疗 人类疾病。

It is proposed that nitric oxide (NO) is a critical messenger and effector molecule in skin physiology and homeostasis and that altered levels of NO in the skin cause disease. The goal of the proposal is to define the role of NO in skin with an emphasis towards determining the functional effects of this gas. Human, but not murine, keratinocytes have been demonstrated to have the capacity to express inducible form of nitric oxide synthase (iNOS) and produce NO. This critical difference between mice and men may explain why mice do not develop cutaneous eruptions analogous to those found in humans. Transgenic mice in which NOS is targeted to the skin mimic are shown here to develop phenotype found in human conditions. In conventional mice, iNOS is expressed in Langerhans cells (LC). NO is also produced by the LC-like cell line XS-52. NO produced in LC may affect LC function and, as NO is freely diffusible across cell membranes, it has the capacity to affect adjacent cells in the epidermis. Toxic effects of NO on melanocytes and keratinocytes suggest that NO may be an effector molecules in a number of skin conditions, including post-inflammatory hypo-pigmentation, vitiligo, graft versus host disease (GVH), scleroderma and the often fatal process, toxic epidermal necrolysis (TEN). In the proposed studies, purified LC will be evaluated for the production of iNOS using RT-PCR and measurement of NO using the Griess reaction and the conversion of radioactive L-arginine to citrulline. The effects of selected cytokines in the regulation of NOS will be examined. The effects of NO on XS-52 cells themselves will be studied. The mechanism of NO- induced killing of melanocytes and keratinocytes will be examined in co- culture experiments with XS-52 cells and via incubation with NO donors. Transgenic mice in which NOS expression is targeted to the epidermis have been produced, phenotypically develop white hair and histopathologically, scleroderma. These animals will be characterized further. Mice expressing NOS under the control of an inducible promoter will be generated. The transgenic mice will be tested as models for disorders of pigmentation, antigen presentation, scleroderma, TEN and GVH. These results have the potential to lead to novel therapeutic strategies for the treatment of human disease.