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Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc BlockTM)

纯化的大鼠抗小鼠CD16 / CD32(小鼠BD Fc Block

Company: BD
Catalog#: 553142
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Isolation and Culture of Mouse Lung ILC2s
Author:
Date:
2018-10-05
[Abstract]  Group 2 Innate Lymphoid Cells (ILC2) play an important role in immune responses at barrier surfaces, notably in the lung during airway allergic inflammation or asthma. Several studies have described methods to isolate ILC2s from wild-type naive mice, most of them using cell sorting to obtain a pure population. Here, we describe in detail, a simple, efficient method for isolation and culture of lung mouse ILC2s. Lungs from Rag2-/- mice pretreated with IL-33 are collected and processed into single cell suspensions. Lymphoid cells are then recovered by density gradient separation. Lin-CD45+ cells are selected by depletion of lineage positive cells followed by positive selection of CD45+ cells. Culture of the isolated cells for several days ... [摘要]  第2组先天性淋巴细胞(ILC2)在屏障表面,特别是在气道过敏性炎症或哮喘期间的肺中的免疫应答中起重要作用。一些研究已经描述了从野生型幼稚小鼠中分离ILC2的方法,其中大多数使用细胞分选来获得纯种群。在这里,我们详细描述了一种简单有效的肺小鼠ILC2分离和培养方法。收集用IL-33预处理的 Rag2 - / - 小鼠的肺并加工成单细胞悬浮液。然后通过密度梯度分离回收淋巴样细胞。通过耗尽谱系阳性细胞然后阳性选择CD45 + 细胞来选择Lin - CD45 + 细胞。将分离的细胞培养数天导致高度纯化的ILC2群体表达典型的细胞表面标志物(CD90.2,Sca1,CD25,CD127和IL-33R)。这些细胞可在培养物中扩增长达10天,并用于多种离体测定或体内过继转移实验。
【背景】第2组先天性淋巴细胞(ILC2)是组织驻留细胞,其在抗寄生虫先天免疫以及过敏性炎症的发展中起关键作用。它们通过产生大量的2型细胞因子IL-5和IL-13对上皮细胞衍生的细胞因子如白细胞介素-33(IL-33)起反应,后者又诱导嗜酸性粒细胞增多和粘液产生(Cayrol和Girard,2018)。为了更好地表征这些细胞的功能和调节,许多组通过荧光激活细胞分选(FACS)从野生型小鼠(WT)的肺中分选ILC2。由于稳定状态下肺中存在的ILC2数量较少,因此该方法导致纯化细胞的产量较低(每只小鼠1×10 ...

Isolation and Analysis of Stromal Cell Populations from Mouse Lymph Nodes
Author:
Date:
2017-08-20
[Abstract]  Our protocol describes a simple procedure for isolating stromal cells from lymph nodes (LN). LN are disrupted then enzymatically digested with collagenase and dispase to produce a single cell suspension that can be stained with fluorescently labelled antibodies and analysed by flow cytometry. This protocol will enable identification of fibroblastic reticular cells (FRC), lymphatic endothelial cells (LEC), blood endothelial cells (BEC) as PNAd+ BEC that form LN high endothelial venules (HEV). This method can be applied to examine LN stromal cell responses during inflammatory events induced by infections or immunologic adjuvants and to subset most leukocytes found in LN. [摘要]  我们的方案描述了从淋巴结(LN)分离基质细胞的简单过程。 LN被破坏,然后用胶原酶和分散酶酶消化以产生可以用荧光标记的抗体染色的单细胞悬浮液并通过流式细胞术分析。 该方案能够识别形成LN高内皮小静脉(HEV)的PNAd + BEC的成纤维细胞网状细胞(FRC),淋巴内皮细胞(LEC),血液内皮细胞(BEC)。 该方法可以用于检测由感染或免疫佐剂诱导的炎症事件期间的LN基质细胞反应,并且在LN中发现大多数白细胞。
【背景】淋巴结(LN)由间充质和内皮基质细胞的复杂网络构成。这些包括成纤维细胞网状细胞(FRCs),淋巴内皮细胞(LECs)和血液内皮细胞(BECs)。这些基质细胞组织了LN的复杂微结构,使得能够支持免疫细胞迁移,体内平衡,耐受性和细胞相互作用,以引发对病原体和肿瘤的免疫应答。我们已经表明,LN基质细胞可以响应于炎症信号而增殖和扩张,并且将免疫细胞募集到伴随感染的LN中(Gregory等,2017)。这些基质细胞也可以显着调节其转录程序以应对感染,从而支持持续的免疫应答。该方案使稳定状态和疾病期间LN的基质细胞亚群可靠地分离。这使得LN基质细胞的表型,功能,遗传或表观遗传学研究揭示了它们如何有助于组织体内平衡和免疫应答。

Mouse CD8+ T Cell Migration in vitro and CXCR3 Internalization Assays
Author:
Date:
2017-03-20
[Abstract]  Chemokines are molecules that regulate the positioning of cells during homeostasis and inflammation. CXCL10 is an interferon-induced chemokine that attracts cells that express the chemokine receptor CXCR3 on their surface. CXCL10 expression is often induced upon inflammation and guides lymphocytes, such as T and NK cells, into the injured tissues. Notably, CXCL10 binding to CXCR3 induces receptor internalization and, therefore, low CXCR3 levels in cells positive for CXCR3 expression can be indicative of chemokine signaling.

Here, we describe an in vitro method to evaluate the ability of murine CD8+ T cells to migrate towards recombinant murine CXCL10; and a flow cytometry assay to measure CXCR3 expression levels at the surface of T cells, after exposure to ...
[摘要]  趋化因子是调节体内平衡和炎症期间细胞定位的分子。 CXCL10是干扰素诱导的趋化因子,其吸引在其表面上表达趋化因子受体CXCR3的细胞。 CXCL10表达通常在炎症诱导并引导淋巴细胞如T和NK细胞进入受损组织。值得注意的是,CXCL10与CXCR3结合诱导受体内化,因此CXCR3表达阳性细胞中的CXCR3水平降低可能是趋化因子信号传导的指示。
 这里,我们描述体外方法来评估鼠CD8 + T细胞向重组鼠CXCL10迁移的能力;以及暴露于不同剂量的趋化因子后在T细胞表面测量CXCR3表达水平的流式细胞术测定。

背景 趋化因子介导的T细胞运输是稳态和炎症期间的重要过程。活化的CD8 + T细胞表达趋化因子受体,例如CXCR3,允许它们向趋化因子CXCL9,10和11迁移,通常在损伤组织上上调。调节T细胞迁移的分子线索的评估对于了解其功能背后的生物学非常重要,但是在体内运行的复杂机制有时难以去卷积。在这里,我们提供有关体外方法的详细信息,以评估CD8 + T细胞上的趋化因子功能,重点是CXCL10介导的化学吸引和CXCR3内化。我们使用可以容易地在体外扩增和活化的抗原特异性转基因CD8 +细胞,因此提供足够数量的表型相同的淋巴细胞(例如, ...

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