ENDOTOXIN RESISTANCE AFTER ADENOVIRAL GENE TRANSFER

描述（来自申请人摘要的逐字记录） 细菌的外膜成分与一种或另一种形式的宿主细胞 在过去几十年里一直是无数调查的对象。的 革兰氏阴性菌的脂多糖内毒素（LPS）是 这些生物体外膜的一种基本元素， 两亲分子可以结合并刺激各种类型的哺乳动物 细胞这种相互作用的结果是多种病理生理学的， 宿主的药理学和免疫学反应。 在过去几年中，我们的长期目标是获得更好的 了解LPS内毒素如何影响宿主细胞。知识 宿主的反应受遗传控制是理解这一点的关键。 因此，我们追求的目标是通过使用 C3 H/HeJ小鼠品系，其细胞具有对LPS的特异性缺陷。本 最后，通过功能性cDNA克隆，我们分离到一个Lps/Ran基因， Ran TC4 GT3我们还从大肠杆菌细胞中分离了Ran LPS（d）cDNA， C3 H/HeJ小鼠，其在cDNA的位置870处具有点突变。这 导致LPS内毒素反应的深刻变化，包括 下调巨噬细胞产生的TNF α，降低B细胞增殖 和促有丝分裂性以及对内毒素攻击的抗性。的这些差异 生物反应是Lpsd/Ran蛋白更快迁移的结果 进入细胞核。我们的具体目标是这些目标的逻辑延伸， 调查结果，以及最近令人兴奋的发现， Toll样受体在先天免疫中的作用，包括对Toll样受体的体内研究， tlr 4和tlr 2及其显性负突变体的生物学效应，以及 将这些效应与LPS/Ran进行比较; tlr 2/tlr 4与 Lps/Ran对内毒素诱导的小鼠致死性损伤的保护作用 几个品系的近交系小鼠; Lpsd/Ran作为 预防性;高表达Lpsd Ran基因的组织或细胞的类型 在对内毒素攻击具有抗性的小鼠中;如果Lpsd/Ran 可以使C3 H/HeOuJ小鼠对LpsD/Ran产生抗性，并且如果LpsD/Ran可以使C3 H/HeJ 小鼠内毒素攻击敏感;和Lps/Ran在其中的作用， 对革兰氏阴性细菌感染的抵抗力。我们相信这些研究将 为临床应用提供有见地的信息。

DESCRIPTION (Verbatim from the applicant's abstract) The interaction of the outer membrane components of bacteria with host cells in one form or another has been the subject of numerous investigations over the past decades. The lipopolysaccharide endotoxin (LPS) of Gram-negative bacteria is an example of an essential element of the outer membrane of these organisms that as an amphipathic molecule can bind to and stimulate various types of mammalian cells. The results of this interaction are multiple pathophysiological, pharmacological and immunological responses by the host. Over the preceding years, our long-term objective has been to gain a better understanding of how LPS endotoxin affects cells of the host. The knowledge that host responses are under genetic control is key to this understanding. Consequently, we have pursued the goal of isolating the LPS gene by the use of the C3H/HeJ mouse strain whose cells possess a specific defect for LPS. To this end, we have isolated a gene Lps/Ran by functional cDNA cloning, which encodes for Ran TC4 GTPase. We have also isolated the Ran Lps(d) cDNA from cells of C3H/HeJ mice, which has a point mutation at position 870 of the cDNA. This leads to profound changes in LPS endotoxin responses, which include down-modulation of TNFa production by macrophages, reduced B cell proliferation and mitogenicity, and resistance to endotoxin challenge. These differences in biological responses are the result of faster migration of the Lpsd/Ran protein into the nucleus. Our specific aims, which are a logical extension of these findings, as well as the recent exciting findings about the involvement of the toll-like receptors in innate immunity, include the studying of the in vivo biological effects of tlr4 and tlr2, with their dominant negative mutants, and compared these effects with Lps/Ran; the relationship between tlr2/tlr4 and Lps/Ran by endotoxin-induced lethality; the protective effect of Lpsd/Ran in several strains of inbred mice; the protective effect of Lpsd/Ran as a prophylactic; the types of tissues or cells highly expressing the Lpsd Ran gene in mice rendered resistant to endotoxin challenge; the examination if Lpsd/Ran could render C3H/HeOuJ mice resistant to, and if LpsD/Ran could render C3H/HeJ mice sensitive to endotoxin challenge; and the role of Lps/Ran in its resistance to Gram-negative bacterial infection. We believe these study will provide insightful information towards clinical application.