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LPS from E. coli, Serotype EH100 (Ra) (TLRgradeTM) (Ready-to-Use)

Company: Enzo Life Sciences
Catalog#: ALX-581-010-L002
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Screening for Novel Endogenous Inflammatory Stimuli Using the Secreted Embryonic Alkaline Phosphatase NF-κB Reporter Assay
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2017-04-05
[Abstract]  An immune response can be activated by pathogenic stimuli, as well as endogenous danger signals, triggering the activation of pattern recognition receptors and initiating signalling cascades that lead to inflammation. This method uses THP1-BlueTM cells, a human monocytic cell line which contains an embryonic alkaline phosphatase reporter gene allowing the detection of NF-κB-induced transcriptional activation. We validated this protocol by assessing NF-κB activation after stimulation of toll-like receptor 4 (TLR4) by two different agonists: lipopolysaccharide (LPS), derived from the cell wall of Gram negative bacteria, and tenascin-C, an extracellular matrix protein whose expression is induced upon tissue injury. We then used this protocol to screen for potential new endogenous ... [摘要]  免疫反应可以被致病性刺激以及内源性危险信号激活,引发模式识别受体的激活并引发导致炎症的信号级联。该方法使用THP1-Blue TM 细胞,其是含有能够检测NF-κB诱导的转录激活的胚胎碱性磷酸酶报告基因的人单核细胞系。通过两种不同的激动剂刺激toll样受体4(TLR4)后,通过评估NF-κB活化来验证该方案:从革兰氏阴性细菌的细胞壁衍生的脂多糖(LPS)和腱生蛋白C,细胞外基质蛋白其组织损伤引起其表达。然后我们使用该方案筛选潜在的新的内源性TLR4激动剂,但是该方法也可以用作快速,经济和可靠的方法来测定导致TLR依赖性NF-κB活化的其它炎性刺激的活性。

免疫系统已经发展成不仅识别病原性刺激物,例如细菌成分和病毒核酸,还包括内源性危险信号,包括从坏死细胞分泌的蛋白质或表达于组织损伤。通过模式识别受体感测到两种类型的刺激,启动触发炎症反应的信号级联。活化的B细胞(NF-κB)的核因子κ-轻链增强子是激活对感染和组织损伤的免疫应答所必需的转录因子。 NF-κB在模式识别受体激活后,在广泛的炎症介质的表达中具有重要作用,包括细胞因子(如肿瘤坏死因子α[TNFα],白细胞介素-6和白细胞介素-1),趋化因子例如白细胞介素-8或CXCL1),蛋白酶,生长因子和MHC相关分子等。为了评估在TLR4下游的该途径的活化,我们使用市售的细胞系THP1-Blue ...

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