FACS-based detection of extracellular ASC specks from NLRP3 inflammasomes in inflammatory diseases
The apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is crucial for inflammasome assembly and activation of several inflammasomes, including NLRP3. ASC aggregates are detected in human sera post pyroptotic cell death, but their inflammasome origin remains unclear...
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| Veröffentlicht in: | Clinical and experimental immunology 2024-12, Vol.219 (1) |
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| container_title | Clinical and experimental immunology |
| container_volume | 219 |
| creator | Topping, Joanne Lara-Reyna, Samuel Ibbotson, Alice Jarosz-Griffiths, Heledd Chang, Leon Poulter, James Peckham, Daniel McDermott, Michael F Savic, Sinisa |
| description | The apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is crucial for inflammasome assembly and activation of several inflammasomes, including NLRP3. ASC aggregates are detected in human sera post pyroptotic cell death, but their inflammasome origin remains unclear.
This study aimed to develop a method to detect ASC aggregates originating from NLRP3 inflammasomes. Initially, human monocytes, macrophages, and THP-1 ASC reporter cells were employed to validate the detection of ASC/NLRP3-positive events through flow cytometry.
The presence of ASC/NLRP3 specks was confirmed in cell supernatants from monocytes and macrophages treated with LPS and nigericin or ATP. Flow cytometry analysis identified double-positive specks in patient sera from inflammatory conditions when compared with healthy controls. Elevated ASC/NLRP3 specks were observed in conditions such as cryopyrin-associated periodic syndrome and Schnitzler's syndrome.
We validated fluorescence-activated cell sorting as a reliable method for detecting ASC/NLRP3 specks in human sera, with potential diagnostic and monitoring applications in certain systemic autoinflammatory diseases. |
| doi_str_mv | 10.1093/cei/uxae117 |
| format | Article |
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This study aimed to develop a method to detect ASC aggregates originating from NLRP3 inflammasomes. Initially, human monocytes, macrophages, and THP-1 ASC reporter cells were employed to validate the detection of ASC/NLRP3-positive events through flow cytometry.
The presence of ASC/NLRP3 specks was confirmed in cell supernatants from monocytes and macrophages treated with LPS and nigericin or ATP. Flow cytometry analysis identified double-positive specks in patient sera from inflammatory conditions when compared with healthy controls. Elevated ASC/NLRP3 specks were observed in conditions such as cryopyrin-associated periodic syndrome and Schnitzler's syndrome.
We validated fluorescence-activated cell sorting as a reliable method for detecting ASC/NLRP3 specks in human sera, with potential diagnostic and monitoring applications in certain systemic autoinflammatory diseases.</description><identifier>ISSN: 0009-9104</identifier><identifier>ISSN: 1365-2249</identifier><identifier>EISSN: 1365-2249</identifier><identifier>DOI: 10.1093/cei/uxae117</identifier><identifier>PMID: 39657685</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>CARD Signaling Adaptor Proteins - metabolism ; Cryopyrin-Associated Periodic Syndromes - diagnosis ; Cryopyrin-Associated Periodic Syndromes - immunology ; Flow Cytometry - methods ; Humans ; Inflammasomes - metabolism ; Inflammation - immunology ; Macrophages - immunology ; Macrophages - metabolism ; Monocytes - immunology ; Monocytes - metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism ; THP-1 Cells</subject><ispartof>Clinical and experimental immunology, 2024-12, Vol.219 (1)</ispartof><rights>The Author(s) 2024. Published by Oxford University Press on behalf of the British Society for Immunology.</rights><rights>The Author(s) 2024. Published by Oxford University Press on behalf of the British Society for Immunology. 2024</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c270t-3e63d761c2fadb691edded90abc90bacb18079158fde63dd319f1deb4b592b843</cites><orcidid>0000-0001-7910-0554</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/39657685$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Topping, Joanne</creatorcontrib><creatorcontrib>Lara-Reyna, Samuel</creatorcontrib><creatorcontrib>Ibbotson, Alice</creatorcontrib><creatorcontrib>Jarosz-Griffiths, Heledd</creatorcontrib><creatorcontrib>Chang, Leon</creatorcontrib><creatorcontrib>Poulter, James</creatorcontrib><creatorcontrib>Peckham, Daniel</creatorcontrib><creatorcontrib>McDermott, Michael F</creatorcontrib><creatorcontrib>Savic, Sinisa</creatorcontrib><title>FACS-based detection of extracellular ASC specks from NLRP3 inflammasomes in inflammatory diseases</title><title>Clinical and experimental immunology</title><addtitle>Clin Exp Immunol</addtitle><description>The apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is crucial for inflammasome assembly and activation of several inflammasomes, including NLRP3. ASC aggregates are detected in human sera post pyroptotic cell death, but their inflammasome origin remains unclear.
This study aimed to develop a method to detect ASC aggregates originating from NLRP3 inflammasomes. Initially, human monocytes, macrophages, and THP-1 ASC reporter cells were employed to validate the detection of ASC/NLRP3-positive events through flow cytometry.
The presence of ASC/NLRP3 specks was confirmed in cell supernatants from monocytes and macrophages treated with LPS and nigericin or ATP. Flow cytometry analysis identified double-positive specks in patient sera from inflammatory conditions when compared with healthy controls. Elevated ASC/NLRP3 specks were observed in conditions such as cryopyrin-associated periodic syndrome and Schnitzler's syndrome.
We validated fluorescence-activated cell sorting as a reliable method for detecting ASC/NLRP3 specks in human sera, with potential diagnostic and monitoring applications in certain systemic autoinflammatory diseases.</description><subject>CARD Signaling Adaptor Proteins - metabolism</subject><subject>Cryopyrin-Associated Periodic Syndromes - diagnosis</subject><subject>Cryopyrin-Associated Periodic Syndromes - immunology</subject><subject>Flow Cytometry - methods</subject><subject>Humans</subject><subject>Inflammasomes - metabolism</subject><subject>Inflammation - immunology</subject><subject>Macrophages - immunology</subject><subject>Macrophages - metabolism</subject><subject>Monocytes - immunology</subject><subject>Monocytes - metabolism</subject><subject>NLR Family, Pyrin Domain-Containing 3 Protein - metabolism</subject><subject>THP-1 Cells</subject><issn>0009-9104</issn><issn>1365-2249</issn><issn>1365-2249</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkUtLxDAUhYMoOj5W7iVLQapJ06aTlQzFUWFQ8bEOedxqtW3GpJWZf28Gx0FXl3PzcZKcg9AxJeeUCHZhoL4YFgooLbbQiDKeJ2maiW00IoSIRFCS7aH9EN6j5Jynu2iPCZ4XfJyPkJ5OyqdEqwAWW-jB9LXrsKswLHqvDDTN0CiPJ08lDnMwHwFX3rX4bvb4wHDdVY1qWxVcCyGqzaJ3foltHSD6hkO0U6kmwNF6HqCX6dVzeZPM7q9vy8ksMWlB-oQBZ7bg1KSVspoLCtaCFURpI4hWRtMxKQTNx5VdkZZRUVELOtO5SPU4Ywfo8sd3PugWrIEu_qCRc1-3yi-lU7X8f9LVb_LVfckYXJ7xjEeH07WDd58DhF62dVhloDpwQ5CMZjE_xtM8omc_qPEuBA_V5h5K5KoWGWuR61oiffL3aRv2twf2DXmdjKE</recordid><startdate>20241209</startdate><enddate>20241209</enddate><creator>Topping, Joanne</creator><creator>Lara-Reyna, Samuel</creator><creator>Ibbotson, Alice</creator><creator>Jarosz-Griffiths, Heledd</creator><creator>Chang, Leon</creator><creator>Poulter, James</creator><creator>Peckham, Daniel</creator><creator>McDermott, Michael F</creator><creator>Savic, Sinisa</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-7910-0554</orcidid></search><sort><creationdate>20241209</creationdate><title>FACS-based detection of extracellular ASC specks from NLRP3 inflammasomes in inflammatory diseases</title><author>Topping, Joanne ; Lara-Reyna, Samuel ; Ibbotson, Alice ; Jarosz-Griffiths, Heledd ; Chang, Leon ; Poulter, James ; Peckham, Daniel ; McDermott, Michael F ; Savic, Sinisa</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c270t-3e63d761c2fadb691edded90abc90bacb18079158fde63dd319f1deb4b592b843</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>CARD Signaling Adaptor Proteins - metabolism</topic><topic>Cryopyrin-Associated Periodic Syndromes - diagnosis</topic><topic>Cryopyrin-Associated Periodic Syndromes - immunology</topic><topic>Flow Cytometry - methods</topic><topic>Humans</topic><topic>Inflammasomes - metabolism</topic><topic>Inflammation - immunology</topic><topic>Macrophages - immunology</topic><topic>Macrophages - metabolism</topic><topic>Monocytes - immunology</topic><topic>Monocytes - metabolism</topic><topic>NLR Family, Pyrin Domain-Containing 3 Protein - metabolism</topic><topic>THP-1 Cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Topping, Joanne</creatorcontrib><creatorcontrib>Lara-Reyna, Samuel</creatorcontrib><creatorcontrib>Ibbotson, Alice</creatorcontrib><creatorcontrib>Jarosz-Griffiths, Heledd</creatorcontrib><creatorcontrib>Chang, Leon</creatorcontrib><creatorcontrib>Poulter, James</creatorcontrib><creatorcontrib>Peckham, Daniel</creatorcontrib><creatorcontrib>McDermott, Michael F</creatorcontrib><creatorcontrib>Savic, Sinisa</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Clinical and experimental immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Topping, Joanne</au><au>Lara-Reyna, Samuel</au><au>Ibbotson, Alice</au><au>Jarosz-Griffiths, Heledd</au><au>Chang, Leon</au><au>Poulter, James</au><au>Peckham, Daniel</au><au>McDermott, Michael F</au><au>Savic, Sinisa</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>FACS-based detection of extracellular ASC specks from NLRP3 inflammasomes in inflammatory diseases</atitle><jtitle>Clinical and experimental immunology</jtitle><addtitle>Clin Exp Immunol</addtitle><date>2024-12-09</date><risdate>2024</risdate><volume>219</volume><issue>1</issue><issn>0009-9104</issn><issn>1365-2249</issn><eissn>1365-2249</eissn><abstract>The apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is crucial for inflammasome assembly and activation of several inflammasomes, including NLRP3. ASC aggregates are detected in human sera post pyroptotic cell death, but their inflammasome origin remains unclear.
This study aimed to develop a method to detect ASC aggregates originating from NLRP3 inflammasomes. Initially, human monocytes, macrophages, and THP-1 ASC reporter cells were employed to validate the detection of ASC/NLRP3-positive events through flow cytometry.
The presence of ASC/NLRP3 specks was confirmed in cell supernatants from monocytes and macrophages treated with LPS and nigericin or ATP. Flow cytometry analysis identified double-positive specks in patient sera from inflammatory conditions when compared with healthy controls. Elevated ASC/NLRP3 specks were observed in conditions such as cryopyrin-associated periodic syndrome and Schnitzler's syndrome.
We validated fluorescence-activated cell sorting as a reliable method for detecting ASC/NLRP3 specks in human sera, with potential diagnostic and monitoring applications in certain systemic autoinflammatory diseases.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>39657685</pmid><doi>10.1093/cei/uxae117</doi><orcidid>https://orcid.org/0000-0001-7910-0554</orcidid><oa>free_for_read</oa></addata></record> |
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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE |
| subjects | CARD Signaling Adaptor Proteins - metabolism Cryopyrin-Associated Periodic Syndromes - diagnosis Cryopyrin-Associated Periodic Syndromes - immunology Flow Cytometry - methods Humans Inflammasomes - metabolism Inflammation - immunology Macrophages - immunology Macrophages - metabolism Monocytes - immunology Monocytes - metabolism NLR Family, Pyrin Domain-Containing 3 Protein - metabolism THP-1 Cells |
| title | FACS-based detection of extracellular ASC specks from NLRP3 inflammasomes in inflammatory diseases |
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