| Quantitative Measurements of HIV-1 and Dextran Capture by Human Monocyte-derived Dendritic Cells (MDDCs)
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Author:
Date:
2016-11-20
[Abstract] The aim of this protocol is to describe how to measure and quantify the amount of HIV-1 particles and dextran molecules internalized in human monocyte derived dendritic cells (MDDCs), using three different techniques: flow cytometry, quantitative PCR and confocal microscopy.
[摘要] 本协议的目的是描述如何使用三种不同的技术：流式细胞术，定量PCR和共聚焦显微镜来测量和量化人类单核细胞衍生树突细胞（MDDCs）中内在的HIV-1颗粒和葡聚糖分子的量。
[背景] 开发此协议是为了评估在人单核细胞衍生的树突细胞中肌动蛋白成核破坏时HIV-1内化的变化。在shRNA筛选后，识别对于HIV-1从树突状细胞转移到T细胞重要的基因，我们观察到肌动蛋白成核的中断导致从富含肌动蛋白的树突到水泡的转变，由于过量的肌动球蛋白收缩。结果，观察到HIV-1转移的减少和由于水泡收缩驱动的巨噬细胞增多引起的HIV-1内化的增加。我们的结论是，肌动蛋白成核和稳定的效应器是维持艾滋病毒1对肌动蛋白丰富的树突和限制其内吞作用，有效转移到T淋巴细胞的关键（Menager和Littman，2016）。
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| C1q Binding to and Uptake of Apoptotic Lymphocytes by Human Monocyte-derived Macrophages
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Author:
Date:
2013-09-05
[Abstract] To characterize macrophage gene expression profiles during the uptake of autologous apoptotic cells, we developed a unique, more physiologic system using primary human monocyte derived macrophages purified via a nonactivating isolation procedure (and in the absence of contaminating platelets, which can release stimulating signals if activated) and autologous lymphocytes as a source of apoptotic cells. The use of autologous cells as the apoptotic target rather than transformed cell lines avoids antigenic stimulation from “nonself” structures at the HLA level but also from “altered self” signals due to the transformation inherent in cell lines.
[摘要] 为了表征在自体凋亡细胞摄取期间的巨噬细胞基因表达谱，我们开发了一种独特的，更生理的系统，使用通过非活化分离程序纯化的原代人单核细胞衍生的巨噬细胞（和在没有污染的血小板，如果激活可以释放刺激信号 ）和自体淋巴细胞作为凋亡细胞的来源。 使用自体细胞作为凋亡靶而不是转化细胞系避免了来自HLA水平上的"非自身"结构的抗原刺激，而且还由于细胞系中固有的转化而导致的"改变的自身"信号。
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