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XenoLight D-luciferin, Potassium salt

Company: PerkinElmer
Catalog#: 122799
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Real-time in vivo Imaging of LPS-induced Local Inflammation and Drug Deposition in NF-κB Reporter Mice
Author:
Date:
2020-08-20
[Abstract]  Wound, biomaterial, and surgical infections are all characterized by a localized and excessive inflammation, motivating the development of in vivo methods focused on the analysis of local immune events. However, current inflammation models, such as the commonly used in vivo models of endotoxin-induced inflammation are based on systemic, usually intraperitoneal, administration of lipopolysaccharide (LPS), causing endotoxin shock. Here, we describe a model of LPS-induced local inflammation in NF-κB-RE-Luc reporter mice. LPS, alone or with added therapeutic substances, is delivered locally via a hydrogel which is deposited subcutaneously, providing a spatially defined environment, enabling in vivo bioimaging analyses of local NF-κB activation. Evaluation of drug ... [摘要]  [摘要 ] 伤口,生物材料和外科手术感染均以局部和过度炎症为特征,从而推动了专注于分析局部免疫事件的体内方法的发展。然而,当前的炎症模型,例如内毒素诱导的炎症的常用体内模型,是基于系统性的,通常是腹膜内注射脂多糖(LPS),引起内毒素休克。在这里,我们描述了LPS诱导局部炎症的模型NF- κ 乙-RE-Luc报告小鼠。LPS单独或与其他治疗物质一起通过水凝胶局部递送, 沉积皮下,提供一个空间限定的环境中,使得能够在体内生物成像本地NF-的分析κ 乙活化。可以在同一只小鼠中纵向分析药物功效的评估,并且使用荧光标记的药物,可以同时分析局部药物沉积,并将其与炎症部位相关联。最后,该方案还可以用于研究药物从局部沉积的凝胶和其他生物材料的保留和系统释放。

[背景 ] 局部过度的TLR应答的原因,有时不相称的炎症,如在不同类型的伤口和生物材料感染的观察。这些伤口并发症延迟了正常的愈合,并增加了严重感染和败血症的风险。考虑到后者,已经开发了败血症和内毒素休克的几种实验模型,其研究了系统性炎症的发展(Lewis 等,2016)。然而,需要从机械学和治疗学角度解决局部炎症事件的模型。甲转录ctivation 因子NF- κ 乙是各种炎性病症的关键组成部分,因此,NF- κ 乙被认为是重要的治疗靶标(刘等人,2017) 。实时的,纵向体内NF-的成像κ 乙活化NF κ ...

Nab Escaping AAV Mutants Isolated from Mouse Muscles
Author:
Date:
2018-05-05
[Abstract]  Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These AAV chimeric capsid mutants enhanced Nab evasion from patient serum with a high muscle transduction efficacy. In this protocol, we describe the procedures for selection of the Nab-escaped AAV chimeric capsid, including isolation and characterization of Nab-escaping AAV mutants in mice muscle. [摘要]  中和抗体(Nabs)是腺相关病毒(AAV)载体基因治疗的临床试验中的主要挑战,因为Nab能够抑制患者中的AAV转导。 我们已经成功地分离了几种新型Nab逃逸的AAV嵌合衣壳,通过给予AAV混洗库和患者血清的混合物。 这些AAV嵌合衣壳突变体增强了来自患有高肌肉转导功效的患者血清的Nab逃避。 在该协议中,我们描述了选择Nab逃逸的AAV嵌合衣壳的程序,包括在小鼠肌肉中分离和鉴定Nab逃逸的AAV突变体。

【背景】腺相关病毒(AAV)载体已被用于许多临床前研究和临床试验。使用AAV基因疗法已经接受了许多疾病的最终治疗。然而,在未来的临床试验中,循环中的中和抗体的存在对AAV载体的应用提出了主要挑战。已经探索过许多方法来逃避Nab的活动。在此,我们描述了从AAV改组库中选择Nab逃逸突变体的定向进化方法。

DNA改组是产生不同突变体的强大策略。通过连续几轮的表型选择,DNA改组文库的特点是质量更高,目标多样化。来自AAV改组文库的衣壳突变体的高通量选择已被用作探索具有靶向特定组织和逃避Nabs的能力的AAV突变体的有前途的策略。然而,大多数这些选择方法只是在体外测试;一些研究甚至使用了兔抗AAV2血清或人静脉注射免疫球蛋白。体内选择衣壳突变体的方法可以提供产生更有效的AAV突变体的平台,所述AAV突变体不仅从患者血清中逃脱Nab,而且增强特定组织中的转导。 ...

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