{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T06:46:25Z","timestamp":1790664385002,"version":"4.1.0"},"reference-count":21,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T00:00:00Z","timestamp":1666224000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hospices Civils de Lyon"},{"name":"Fondation HCL"},{"name":"Claude Bernard Lyon 1 University"},{"name":"R\u00e9gion Auvergne Rh\u00f4ne-Alpes"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Alteration of NLRP3 inflammasome pathway including hyper-activation or exhaustion has been implicated in the pathophysiology of many diseases. Following cell stimulation, aggregation of the ASC protein into a multiprotein complex, the ASC speck, has been proposed as a specific read-out for monitoring NLRP3 inflammasome activation by flow cytometry in clinical samples. So far, only a few papers have described a technique to detect ASC speck formation directly in whole blood without any cell purification, and none included an ex vivo stimulation. The objective of this study was thus to develop a simple and shortened flow cytometry protocol to detect ASC speck formation directly in whole blood including an ex vivo stimulation step. We showed that after red blood cells lysis and removal of the LPS stimulation step, ASC speck formation can be detected in both monocytes and neutrophils from healthy donors directly in nigericin-stimulated whole blood samples. Using samples from four septic shock patients, we showed that this technique allows for the detection of NLRP3 inflammasome exhaustion in clinical samples. This novel shortened and simple whole blood protocol should facilitate day-to-day monitoring of NLRP3 inflammasome activation and exhaustion in both monocytes and neutrophils in clinical studies.<\/jats:p>","DOI":"10.3390\/cells11203306","type":"journal-article","created":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T20:35:55Z","timestamp":1666298155000},"page":"3306","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Monitoring NLRP3 Inflammasome Activation and Exhaustion in Clinical Samples: A Refined Flow Cytometry Protocol for ASC Speck Formation Measurement Directly in Whole Blood after Ex Vivo Stimulation"],"prefix":"10.3390","volume":"11","author":[{"given":"R\u00e9my","family":"Coudereau","sequence":"first","affiliation":[{"name":"Immunology Laboratory, Edouard Herriot Hospital, Hospices Civils de Lyon, 5 Place d\u2019Arsonval, CEDEX 03, 69437 Lyon, France"},{"name":"EA 7426 \u201cPathophysiology of Injury-Induced Immunosuppression\u201d (Universit\u00e9 Claude Bernard Lyon 1\u2014Hospices Civils de Lyon\u2014bioM\u00e9rieux), Joint Research Unit HCL-bioM\u00e9rieux, 69003 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morgane","family":"Gossez","sequence":"additional","affiliation":[{"name":"Immunology Laboratory, Edouard Herriot Hospital, Hospices Civils de Lyon, 5 Place d\u2019Arsonval, CEDEX 03, 69437 Lyon, France"},{"name":"CIRI, Centre International de Recherche en Infectiologie, Univ Lyon, Inserm U1111, Universit\u00e9 Claude Bernard-Lyon 1, CNRS, UMR5308, ENS de Lyon, 69007 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6716-7807","authenticated-orcid":false,"given":"B\u00e9n\u00e9dicte F","family":"Py","sequence":"additional","affiliation":[{"name":"CIRI, Centre International de Recherche en Infectiologie, Univ Lyon, Inserm U1111, Universit\u00e9 Claude Bernard-Lyon 1, CNRS, UMR5308, ENS de Lyon, 69007 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0687-8565","authenticated-orcid":false,"given":"Thomas","family":"Henry","sequence":"additional","affiliation":[{"name":"CIRI, Centre International de Recherche en Infectiologie, Univ Lyon, Inserm U1111, Universit\u00e9 Claude Bernard-Lyon 1, CNRS, UMR5308, ENS de Lyon, 69007 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne-Claire","family":"Lukaszewicz","sequence":"additional","affiliation":[{"name":"EA 7426 \u201cPathophysiology of Injury-Induced Immunosuppression\u201d (Universit\u00e9 Claude Bernard Lyon 1\u2014Hospices Civils de Lyon\u2014bioM\u00e9rieux), Joint Research Unit HCL-bioM\u00e9rieux, 69003 Lyon, France"},{"name":"Anaesthesia and Critical Care Medicine Department, Edouard Herriot Hospital, Hospices Civils de Lyon, 5 Place d\u2019Arsonval, CEDEX 03, 69437 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9961-5739","authenticated-orcid":false,"given":"Guillaume","family":"Monneret","sequence":"additional","affiliation":[{"name":"Immunology Laboratory, Edouard Herriot Hospital, Hospices Civils de Lyon, 5 Place d\u2019Arsonval, CEDEX 03, 69437 Lyon, France"},{"name":"EA 7426 \u201cPathophysiology of Injury-Induced Immunosuppression\u201d (Universit\u00e9 Claude Bernard Lyon 1\u2014Hospices Civils de Lyon\u2014bioM\u00e9rieux), Joint Research Unit HCL-bioM\u00e9rieux, 69003 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0462-4235","authenticated-orcid":false,"given":"Fabienne","family":"Venet","sequence":"additional","affiliation":[{"name":"Immunology Laboratory, Edouard Herriot Hospital, Hospices Civils de Lyon, 5 Place d\u2019Arsonval, CEDEX 03, 69437 Lyon, France"},{"name":"CIRI, Centre International de Recherche en Infectiologie, Univ Lyon, Inserm U1111, Universit\u00e9 Claude Bernard-Lyon 1, CNRS, UMR5308, ENS de Lyon, 69007 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"359","DOI":"10.2147\/JIR.S141220","article-title":"Spotlight on the NLRP3 inflammasome pathway","volume":"11","author":"Groslambert","year":"2018","journal-title":"J. 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