| 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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| Determination of Toxoplasma gondii Replication in Naïve and Activated Macrophages
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Author:
Date:
2012-11-20
[Abstract] Toxoplasma gondii is an obligate intracellular protozoan parasite that causes the disease toxoplasmosis. Chronic infection is established through the formation of tissue cysts predominantly in cardiac and neurologic tissues. A defining characteristic of T. gondii is its ability to evade the host’s immune defenses; specifically, T. gondii can invade and persist within host phagocytes, using them to disseminate to the brain and central nervous system where cysts are then formed. This protocol is used to evaluate the ability of Toxoplasma gondii to survive and replicate within naive and activated murine bone marrow-derived macrophages at the level of single infected cells. In the following protocol macrophages are naive or activated with IFN-γ and LPS but ...
[摘要] 弓形虫是一种专性的细胞内原生动物寄生虫，其引起疾病弓形体病。慢性感染通过主要在心脏和神经组织中形成组织囊肿来建立。 T的定义特性。 gondii 是其逃避宿主免疫防御的能力;具体地，em。 gondii可以侵入并持续在宿主吞噬细胞内，使用它们传播到脑和中枢神经系统，然后形成囊肿。该方案用于评估弓形虫在单个感染细胞水平下在幼稚和活化的鼠骨髓衍生的巨噬细胞中存活和复制的能力。在以下方案中，巨噬细胞是幼稚的或用IFN-γ和LPS活化的，但是可以利用不同的激活刺激以及不同的宿主细胞群体和不同的抑制剂。寄生虫复制通过使用针对寄生虫和显微镜分析的免疫荧光染色评估每个液泡随时间的寄生虫数量来确定。每个液泡的寄生虫数的动力学测定准确地反映了随时间的寄生虫复制，因为含有空泡的寄生虫不彼此融合。分离鼠骨髓来源的巨噬细胞，制备用于收集巨噬细胞集落刺激因子的条件性L929细胞，并且包括在方案中的荧光显微镜染色具有广泛的适用性。该协议对于存在于不彼此融合并且可通过显微镜可视化的液泡中的病原体如弓形体病毒（inxoplasma gondii）起作用。
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