Direct Reprogramming of Mouse Embryonic Fibroblasts to Conventional Type 1 Dendritic Cells by Enforced Expression of Transcription Factors
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Author:
Date:
2020-05-20
[Abstract] Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on antigen cross-presentation, a critical step for inducing CD8+ T cell cytotoxic responses. The rarity of naturally occurring cDC1s and lack of in vitro methodologies for the generation of pure cDC1 populations strongly hinders the ...
[摘要] [摘要] 转录因子组合的异位表达最近被证明可以将分化的体细胞重编程为树突状细胞(DC)谱系,而不会回复到多能状态。DC具有诱导有效和持久的适应性免疫应答的能力。在特定的常规类型1的DC(cDC1s)练成抗原交叉呈递,用于诱导CD8的关键步骤+ T细胞的细胞毒性应答。天然存在的cDC1的稀有性和缺乏用于生成纯cDC1群体的体外方法论,严重阻碍了对cDC1谱系规格和功能的研究。在这里,我们描述了用于生成感应DC(iDC)的协议 慢病毒介导的转录因子PU.1,IRF8和BATF3在小鼠胚胎成纤维细胞中的表达。iDC 在9天内获得DC形态,cDC1表型和转录特征。使用此协议生成的iDC 具有对炎症刺激,吞噬死细胞,将抗原加工并交叉呈递给CD8 + T细胞的功能。DC重新编程提供了一个简单易处理的系统,可以生成大量的cDC1类细胞用于高内涵筛选,从而开辟了新途径,可以更好地了解cDC1的规格和功能。将来,在成纤维细胞中忠实诱导cDC1命运可能会导致产生患者特定的疫苗接种DC。
[背景技术树突状细胞(DC)是专业的抗原呈递细胞,专门用于识别,加工和呈递T细胞抗原,在诱导适应性免疫应答和免疫记忆中起关键作用(Me rad 等,2013)。DC可以分为4个主要子集:浆细胞样DC(pDC ),大量1型干扰素的产生者,循环单核细胞衍生的单核细胞衍生DC 和常规DC(cDC ...
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Mechanical Allodynia Assessment in a Murine Neuropathic Pain Model
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Author:
Date:
2018-01-20
[Abstract] Experimental animal models are unique tools (i) to study pain transmission and pathophysiology of neuropathic pain, (ii) to identify novel molecular targets and (iii) to test the potential analgesic effect of specific molecules. The chronic constriction injury (CCI) model of neuropathic pain is the first model of post-traumatic painful peripheral neuropathy, originally developed by Bennett and Xie in the late 1980s. The chronic constriction is performed in the sciatic nerve and induces a partial denervation involving myelinated afferent axons and unmyelinated axons. Damage to unmyelinated ...
[摘要] 实验动物模型是独特的工具:(i)研究神经性疼痛的疼痛传递和病理生理学,(ii)鉴定新的分子靶标和(iii)测试特定分子的潜在镇痛作用。神经性疼痛的慢性缩窄性损伤(CCI)模型是最初由Bennett和Xie在二十世纪八十年代后期开发的创伤后疼痛性周围神经病的第一个模型。慢性收缩是在坐骨神经中进行的,并诱导部分去神经支配,包括有髓鞘的传入轴突和无髓鞘轴突。对无髓鞘轴突的损伤比有髓神经传入者严重得多。由于该模型导致部分去神经支配,对疼痛行为的分析非常有用。刺激后爪(坐骨神经的靶标)引起可被定量的疼痛。因此,通常通过测量对von Frey丝刺激的后爪缩回反应,在坐骨神经的CCI后7,14和21天评估机械异常性疼痛。在这里,我们详细描述协议允许在小鼠中可靠和可重复的CCI模型。总的来说,研究人员最常使用这种手术模式来发现更有效的慢性疼痛状态的药物控制药物。
【背景】Bennett和Xie(1988)首先提出了神经性疼痛的慢性缩窄性损伤(CCI)模型。慢性缩窄应用于模拟创伤后疼痛性周围神经病的坐骨神经。该模型诱导了部分去神经支配,因此对定量分析疼痛行为和评价新药的镇痛效果非常有用。坐骨神经的CCI在异氟烷麻醉下进行(诱导5%,维持2%)。通过解剖将股二头肌和股浅筋膜分开以暴露坐骨神经。通过在坐骨神经周围松散地结扎一根结扎物来诱导CCI,以保持神经外循环。 ...
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