ENDOTOXIN RESISTANCE AFTER ADENOVIRAL GENE TRANSFER

描述(逐字摘自申请人的摘要) 细菌的外膜成分，宿主细胞以这样或那样的形式存在 在过去的几十年里一直是无数调查的对象。这个 革兰氏阴性菌的脂多糖内毒素就是一个例子 这些生物体外膜的基本成分，作为一种 两亲性分子可以结合和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的， 宿主的药理学和免疫学反应。 在过去的几年里，我们的长期目标一直是获得更好的 了解内毒素如何影响宿主细胞。《知识》 寄主反应受基因控制是理解这一点的关键。 因此，我们一直致力于通过使用 C3H/HeJ小鼠品系，其细胞对内毒素具有特异性缺陷。对这件事 最后，我们通过功能克隆的方法克隆了一个编码Lps/ran的基因 RAN TC4 GTP酶。我们还从小鼠肾小管上皮细胞中分离了Ran Lps(D)基因。 C3H/HeJ小鼠，其cDNA870位点突变。这 导致内毒素反应的深刻变化，包括 巨噬细胞下调TNFa的产生，减少B细胞的增殖 和有丝分裂活性，以及对内毒素攻击的抵抗力。这些差异体现在 生物反应是LPSD/RAN蛋白更快迁移的结果 进入原子核。我们的具体目标，这是这些目标的逻辑延伸 发现，以及最近关于参与的令人兴奋的发现 天然免疫中的Toll样受体，包括体内研究 TLR4和TLR2及其显性负突变体的生物学效应 与LPs/RAN比较，TLR2/TLR4与LPS/RAN的关系 内毒素诱导的LPS/RAN致死性；LPSD/RAN对大鼠的保护作用 几个品系的近交系小鼠；LPSD/RAN作为一种保护作用 预防性；高表达LPSDRAN基因的组织或细胞类型 在内毒素攻击耐受的小鼠中；LPSD/RAN的检查 能使C3H/HeOuJ小鼠产生耐药性，如果LpsD/RAN能使C3H/HeJ 对内毒素攻击敏感的小鼠；以及LPs/RAN在其 对革兰氏阴性细菌感染的抵抗力。我们相信这些研究将会 为临床应用提供有洞察力的信息。

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.