| Quantification of Tumor Material Uptake
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Author:
Date:
2016-10-20
[Abstract] Extracellular tumor material including exosomes, microvesicles and apoptotic tumor debris may help cancers invade new organs. Enhancing the removal of extracellular tumor material by immune cells represents a novel immunotherapy approach for preventing cancer metastasis. This protocol quantifies the uptake and removal of extracellular tumor material from circulation and tissues by immune cells. In this assay fluorescent tumor cells are transferred into mice, and then immune cells are quantified by either flow cytometry or imaging cytometry for their uptake of tumor material.
[摘要] 包括外来体，微泡和凋亡性肿瘤碎片的细胞外肿瘤材料可以帮助癌症侵入新器官。增强免疫细胞对细胞外肿瘤材料的去除代表了用于预防癌症转移的新型免疫治疗方法。该方案定量免疫细胞从循环和组织吸收和去除细胞外肿瘤物质。在该测定中，将荧光肿瘤细胞转移到小鼠中，然后通过流式细胞术或成像细胞计量术来定量免疫细胞对肿瘤材料的摄取。
[背景] 研究已经证明，包括从肿瘤脱落的外来体，微泡和凋亡性肿瘤碎片的细胞外肿瘤材料是肿瘤转移，生长和逃避免疫应答的重要介质（Vader等人，2014; Pucci和Pittet ，2013; Pucci等人，2016）。免疫细胞具有去除，应答和转运这种循环肿瘤材料的能力（Hanna等人，2015; Pucci等人，2016; Headley等人。，2016）。该协议提供了一种新的方法来量化特定免疫细胞群体的肿瘤材料摄取，并且可以适于测试调节肿瘤材料摄取的免疫靶标。该协议可以帮助更好地理解对细胞外肿瘤材料的免疫应答，希望最终开发针对癌症发展中的细胞外肿瘤材料的新疗法。
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| Flow Cytometry of Lung and Bronchoalveolar Lavage Fluid Cells from Mice Challenged with Fluorescent Aspergillus Reporter (FLARE) Conidia
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Author:
Date:
2016-09-20
[Abstract] Aspergillus fumigatus is a ubiquitous fungal pathogen that forms airborne conidia. The process of restricting conidial germination into hyphae by lung leukocytes is critical in determining infectious outcomes. Tracking the outcome of conidia-host cell encounters in vivo is technically challenging and an obstacle to understanding the molecular and cellular basis of antifungal immunity in the lung. Here, we describe a method that utilizes a genetically engineered Aspergillus strain [called FLARE (Jhingran et al., 2012; Espinosa et al., 2014; Heung et ...
[摘要] Aspergillus fumigatus is a ubiquitous fungal pathogen that forms airborne conidia. The process of restricting conidial germination into hyphae by lung leukocytes is critical in determining infectious outcomes. Tracking the outcome of conidia-host cell encounters in vivo is technically challenging and an obstacle to understanding the molecular and cellular basis of antifungal immunity in the lung. Here, we describe a method that utilizes a genetically engineered Aspergillus strain [called FLARE (Jhingran et al., 2012; Espinosa et al., 2014; Heung et al., 2015)] to monitor conidial phagocytosis ...
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| In vitro Assessment of Immunological Synapse Formation by Flow Cytometry
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Author:
Date:
2016-03-20
[Abstract] In adaptive immune system, formation of immunological synapse between T cells and antigen presenting cells (dendritic cells, B cells, and macrophages) or target cells (tumor cells and viral-infected cells) is critical for the execution of T cell immune responses via cytokine secretion or direct killing activity. Here, we describe the practical methods that directly measure the number of conjugates as a result of immunological synapse formation between T cells and superantigen-loaded B cells or between cytotoxic T cells and antigen-loaded target cells by dual-color flow cytometry.
[摘要] 在适应性免疫系统中，T细胞和抗原呈递细胞（树突细胞，B细胞和巨噬细胞）或靶细胞（肿瘤细胞和病毒感染细胞）之间的免疫突触的形成对通过细胞因子执行T细胞免疫应答至关重要 分泌或直接杀死活动。 在这里，我们描述了通过双色流式细胞术直接测量T细胞和加载超抗原的B细胞之间或细胞毒性T细胞和抗原负载的靶细胞之间的免疫突触形成的结合物数量的实际方法。
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