MOLECULAR RESPONSES OF MACROPHAGES TO LIPOPOLYSACCHARIDE

描述（改编自申请人摘要）：细菌LPS是一种 全身炎症反应毒性表现的主要因素 综合症，在美国死亡的主要原因。 炎性细胞因子 由巨噬细胞产生的TNF-α、IL-1和IL-6是巨噬细胞增殖的主要介质。 LPS的毒性作用。 这些研究的目的是了解 LPS诱导的白细胞反应的机制，特别是 参与巨噬细胞对LPS的早期反应的基因产物。 这些研究将使用两种工具，1）同类小鼠品系， C3H/HeN和C3H/HeJ对LPS的反应不同，这是由于一个突变导致的。 单基因，Lps. 2)紫杉醇（一种微管结合剂）模拟 巨噬细胞中的LPS。 建议确定是否有共享 紫杉醇和LPS的细胞靶点，并定义这些靶点及其 在LPS诱导的信号传导中的作用。 还建议确定是否有 是HeN和HeJ小鼠之间差异表达的组成型基因， 并确定该假定基因的作用及其与 LPS基因产物。 为了实现这些目标，研究者将：1） 在无细胞系统中表征巨噬细胞对LPS的早期反应， 为了测试可能参与早期癌症的候选蛋白质的作用， LPS信号传导。 2)鉴定和克隆紫杉醇和紫杉醇共有的细胞靶点， 用标记的紫杉醇筛选巨噬细胞cDNA表达文库， 抗紫杉醇抗体 3)鉴定Lpsn中差异表达的基因 (LPS正常反应）和Lpsd（脂蛋白低反应）细胞使用差异 显示分析，克隆这些基因，并研究它们在 LPS信号传导。

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.