| Measurement of Dipeptidylpeptidase Activity in vitro and in vivo
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Author:
Date:
2017-03-20
[Abstract] Dipeptidylpeptidases (DPPs) are serine proteases, which cleave small proteins and peptides possessing a proline or an alanine in the second position of their N-terminus. Among the members of this family, dipeptidylpeptidase 4 (DPP4) is constitutively expressed in the extracellular space. DPP4 is found at the surface of many hematopoietic and non-hematopoietic cells and is also present in many biological fluids in a bioactive soluble form. DPP4 expression is modulated by inflammation, and measurements of its activity have been used as biomarker for disease. Here, we describe a method to evaluate the enzymatic activity of DPP4 in vitro and in vivo.
[摘要] 二肽基肽酶（DPPs）是丝氨酸蛋白酶，其在其N末端的第二位置切割具有脯氨酸或丙氨酸的小蛋白质和肽。在该家族成员中，二肽基肽酶4（DPP4）在细胞外空间中组成型表达。 DPP4存在于许多造血细胞和非造血细胞的表面，并且也以生物活性可溶形式存在于许多生物流体中。 DPP4表达受炎症调节，其活性的测定已被用作疾病的生物标志物。在这里，我们描述了一种评估体外DPP4和/或体内酶活性的方法。
背景 DPPs的酶活性的大小可以是炎症的指标;以及用于DPP4抑制剂的重要药效学参数。在这里，我们报告了市售试剂盒用于评估体外DPP-酶活性的详细信息;并提供测试生物样品生成的细节。此外，我们描述了该方法如何用于体内DPP活性的评估。
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| 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 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。
＆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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