| Mouse CD8+ T Cell Migration in vitro and CXCR3 Internalization Assays
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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 ...
[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。
＆nbsp;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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| Isolation of CNS-infiltrating and Resident Microglial Cells
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Author:
Date:
2015-01-20
[Abstract] Variety of immunological and biochemical studies associated with infection or inflammation in the central nervous system (CNS) utilize CNS-resident and/or infiltrating cells which were isolated from the CNS of naïve and affected mice in order to investigate the underlying mechanisms and the potential roles of the cell populations. Mechanical and enzyme-based single cell preparations of CNS cells are subjected to a density gradient to obtain functional single cells. In combination with cell-specific biomarkers, the function and/or status of resident microglia and infiltrating lymphocytes, ...
[摘要] 与中枢神经系统（CNS）中的感染或炎症相关的免疫学和生物化学研究的多样性利用从初始和受影响的小鼠的CNS中分离的CNS驻留和/或浸润细胞，以研究潜在的机制和潜在的作用 的细胞群体。 将CNS细胞的机械和基于酶的单细胞制剂进行密度梯度以获得功能性单细胞。 与细胞特异性生物标志物组合，可以表征驻留小胶质细胞和浸润淋巴细胞（包括B和T细胞以及巨噬细胞）的功能和/或状态。
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| Whole Spleen Flow Cytometry Assay
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Author:
Date:
2013-08-05
[Abstract] In the Whole Spleen Flow Cytometry Assay, we used splenocytes directly ex vivo for stimulation with a variety of TLR ligands. The splenocytes were stimulated for a total of 4 hours, then stained for intracellular cytokines. We then examined cytokine production via flow cytometry. This allowed us to compare the responses of minimally manipulated primary macrophages/monocytes and conventional dendritic cells.
[摘要] 在全脾流式细胞术测定中，我们直接使用脾细胞离体用各种TLR配体刺激。 将脾细胞刺激总共4小时，然后对细胞内细胞因子染色。 然后我们通过流式细胞术检查细胞因子产生。 这使我们能够比较最小操纵的原代巨噬细胞/单核细胞和常规树突状细胞的反应。
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