MOLECULAR RESPONSES OF MACROPHAGES TO LIPOPOLYSACCHARIDE

描述(改编自申请人的摘要)：细菌内毒素是一种 全身炎症反应毒性表现的主要因素 综合症，在美国是主要的死亡原因。炎性细胞因子 巨噬细胞产生的肿瘤坏死因子-α、白介素1和白介素6是 内毒素的毒性作用。拟议研究的目标是了解 内毒素诱导白细胞反应的机制，特别是 参与巨噬细胞对内毒素早期反应的基因产物。 这些研究将使用两种工具，1)同源小鼠品系， C3H/HEN和C3H/HEJ对内毒素的反应不同，这是由于 单基因，LPs。2)紫杉醇(一种微管结合剂)模拟 巨噬细胞内的内毒素。建议确定是否存在共享的 紫杉醇和脂多糖的细胞靶点，并定义这些靶点和它们的 在脂多糖诱导的信号转导中的作用。还建议确定是否存在 是一个在母鸡和HeJ小鼠之间差异表达的构成基因， 并确定该假定基因的作用及其与 脂多糖基因产物。为了实现这些目标，调查员将：1) 巨噬细胞在无细胞系统中对内毒素的早期反应 为了测试可能参与早期反应的候选蛋白质的作用 LP信令。2)鉴定和克隆紫杉醇和 用标记紫杉醇和脂多糖筛选巨噬细胞表达文库 抗紫杉醇抗体。3)确定在Lpsn中差异表达的基因 (内毒素正常反应)和LPSD(内毒素低反应)细胞 展示分析，克隆这些基因并研究它们在 LP-信令。

DESCRIPTION (Adapted from the applicant's abstract): Bacterial LPS is a major factor in the toxic manifestations of systemic inflammatory response syndrome, a major cause of death in the US. The inflammatory cytokines TNF-alpha, IL-1 and IL-6 are produced by macrophages are major mediators of the toxic effects of LPS. The goal of the proposed studies is to understand the mechanisms of LPS-induced responses in leukocytes, in particular the gene products participating in the early response of macrophages to LPS. Two tools will be used for these studies, 1) the congenic mouse strains, C3H/HeN and C3H/HeJ, which respond differently to LPS due to a mutation in a single gene, Lps. 2) Taxol (a microtubule-binding agent) mimics actions of LPS in macrophages. It is proposed to determine if there are shared cellular targets of taxol and LPS, and to define these targets and their roles in LPS-induced signaling. It is also proposed to determine if there is a constitutive gene differentially expressed between HeN and HeJ mice, and to determine the role of this putative gene and its relationship to the LPS gene product. To accomplish these goals, the investigator will: 1) Characterize early responses of macrophages to LPS in a cell free system in order to test the role of candidate proteins that might participate in early LPS signaling. 2) Identify and clone cellular targets shared by taxol and LPS by screening a macrophage cDNA expression library with labeled taxol and anti-taxol antibody. 3) Identify genes differentially expressed in Lpsn (LPS normoresponsive) and Lpsd (LPS-hyporesponsive) cells using differential display analysis, to clone these genes and study their roles in LPS-signaling.