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{"title":"Monitoring Inflammasome Priming and Activation in Response to Candida albicans","authors":"Darian J. Santana, Faith M. Anderson, Teresa R. O'Meara","doi":"10.1002/cpmc.124","DOIUrl":null,"url":null,"abstract":"<p><i>Candida albicans</i> is a common mucosal colonizer, as well as a cause of lethal invasive fungal infections. The major predisposing factor for invasive fungal disease is a compromised immune system. One component of the host immune response to fungal infection is the activation of the inflammasome, a multimeric protein complex that is critical for regulating host pro-inflammatory responses. Here, we describe methods for investigating the interactions between <i>C. albicans</i> and host macrophages, with a focus on the inflammasome. <i>C. albicans</i> isolates differ in the degree to which they activate the inflammasome due to differences in internalization, morphogenic switching, and inflammasome priming. Therefore, we include protocols for identifying these factors. This simple in vitro model can be used to elucidate the contributions of specific <i>C. albicans</i> strains or mutants to different aspects of interactions with macrophages. © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Measuring inflammasome priming in response to <i>Candida albicans</i></p><p><b>Basic Protocol 2</b>: Measuring inflammasome activation in response to <i>Candida albicans</i></p><p><b>Support Protocol</b>: Controlling for phagocytosis</p>","PeriodicalId":39967,"journal":{"name":"Current Protocols in Microbiology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2020-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmc.124","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.124","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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Abstract
Candida albicans is a common mucosal colonizer, as well as a cause of lethal invasive fungal infections. The major predisposing factor for invasive fungal disease is a compromised immune system. One component of the host immune response to fungal infection is the activation of the inflammasome, a multimeric protein complex that is critical for regulating host pro-inflammatory responses. Here, we describe methods for investigating the interactions between C. albicans and host macrophages, with a focus on the inflammasome. C. albicans isolates differ in the degree to which they activate the inflammasome due to differences in internalization, morphogenic switching, and inflammasome priming. Therefore, we include protocols for identifying these factors. This simple in vitro model can be used to elucidate the contributions of specific C. albicans strains or mutants to different aspects of interactions with macrophages. © 2020 Wiley Periodicals LLC.
Basic Protocol 1 : Measuring inflammasome priming in response to Candida albicans
Basic Protocol 2 : Measuring inflammasome activation in response to Candida albicans
Support Protocol : Controlling for phagocytosis
监测炎性体对白色念珠菌的启动和激活
白色念珠菌是一种常见的粘膜定植菌,也是致死性侵袭性真菌感染的原因。侵袭性真菌疾病的主要诱发因素是免疫系统受损。宿主对真菌感染的免疫反应的一个组成部分是炎症小体的激活,炎症小体是一种多聚体蛋白复合物,对调节宿主的促炎反应至关重要。在这里,我们描述了研究白色念珠菌和宿主巨噬细胞之间相互作用的方法,重点是炎性体。由于内化、形态发生转换和炎性小体启动的差异,白色念珠菌分离株在激活炎性小体的程度上有所不同。因此,我们包含了识别这些因素的协议。这个简单的体外模型可以用来阐明特定的白色念珠菌菌株或突变体在与巨噬细胞相互作用的不同方面的贡献。©2020 Wiley期刊有限公司。基本方案1:测量白色念珠菌反应的炎症小体启动。基本方案2:测量白色念珠菌反应的炎症小体激活。支持方案:控制吞噬
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