| In vitro Differentiation of Murine Innate Lymphoid Cells from Common Lymphoid Progenitor Cells
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Author:
Date:
2016-03-20
[Abstract] Subtypes of innate lymphoid cells (ILC), defined based on their cytokine secretion profiles and transcription factor expression, are important for host protection from pathogens and maintaining tissue homeostasis. ILCs develop from common lymphoid progenitors (CLP) in the bone marrow. Using the methods described here, we have previously shown that loss of the transcriptional regulator TOX (Thymocyte-selection associated HMG-box protein) leads to specific changes in ILC development and differentiation. Here, we describe how to obtain ILCs from in vivo isolated CLP grown in vitro.
[摘要] 先天淋巴细胞（ILC）的亚型，基于其细胞因子分泌谱和转录因子表达来定义，对于宿主保护免于病原体和维持组织内稳态是重要的。 ILC从骨髓中的普通淋巴祖细胞（CLP）发展而来。 使用本文所述的方法，我们以前显示转录调节器TOX（胸腺细胞选择相关的HMG框蛋白）的损失导致ILC发展和分化的具体变化。 在这里，我们描述了如何从体内分离的CLP获得ILC。
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| Isolation and in vivo Transfer of Antigen Presenting Cells
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Author:
Date:
2014-10-05
[Abstract] Transfer of antigen presenting cells in vivo is a method used by immunologists to examine the potency of antigen presentation by a selected population of cells. This method is most commonly used to analyze presentation of protein antigens to MHC class I or II restricted T cells, but it can also be used for studies of nonconventional antigens such as CD1-presented lipids. In a recent study focusing on CD1d-restricted glycolipid antigen presentation to Natural Killer T cells, we compared antigen presenting properties of splenic B cells, CD8αPos dendritc cells (DCs) and CD8αNeg DCs (Arora et al., 2014). This protocol describes the detailed method used for isolation of these cell populations, and their transfer into recipient mice to analyze their ...
[摘要] 体内转移抗原呈递细胞是免疫学家用来检查所选择的细胞群体的抗原呈递效力的方法。该方法最常用于分析蛋白质抗原对MHC I类或II类限制性T细胞的呈递，但其也可用于非常规抗原如CD1呈递脂质的研究。在最近关于CD1d限制性糖脂抗原呈递到天然杀伤T细胞的研究中，我们比较脾B细胞，CD8αpos 树突细胞（DC）和CD8α Neg sup> DC（Arora et al。，2014）。该方案描述了用于分离这些细胞群体和将其转移到受体小鼠中以分析其抗原呈递性质的详细方法。作为总单核细胞的百分比，平均脾脏含有约1-3％骨髓树突状细胞（DC）。在绝对数量上，这转换为大约0.6-1.8×10 6个DC。为了增强脾中DC的数目，向小鼠皮下注射来自经过工程改造以表达fms相关的酪氨酸激酶3配体（Flt3L）的培养的黑素瘤细胞系（B16.Flt3L）的细胞（Mach等人。，2000）。该蛋白是与集落刺激因子-1同源的生长因子，并在造血干细胞的分化中起关键作用。将这种蛋白质作为纯化的蛋白质施用到小鼠中导致多个器官中的CD8αpos pos和CD8α阳性DC亚群的扩增。在已经植入过表达这种蛋白质的肿瘤细胞的小鼠中也观察到类似的扩增（Mach等人，2000）。根据我们的经验，来自具有可触知的B16.Flt3L肿瘤的小鼠的脾脏中高达60％的总单核细胞可以是CD11c阳性树突细胞，从而得到高达5×10 ...
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| Crosslinking and Immunoprecipitation in Zebrafish
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Author:
Date:
2012-01-20
[Abstract] Immunoprecipitation (IP) is a routine method to detect protein binding and interactions. But the weak binding between two proteins is often hard to detect during regular IP procedure. This protocol offers a crossliking and IP combination method to detect weak binding of proteins in zebrafish embryos.
[摘要] 免疫沉淀（IP）是检测蛋白质结合和相互作用的常规方法。 但两个蛋白质之间的弱绑定往往很难在常规IP过程中检测。 这个协议提供crossliking和IP组合方法来检测斑马鱼胚胎中蛋白质的弱绑定。
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