{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,14]],"date-time":"2026-03-14T14:03:14Z","timestamp":1773496994089,"version":"3.50.1"},"reference-count":28,"publisher":"Wiley","issue":"20","license":[{"start":{"date-parts":[[2021,8,28]],"date-time":"2021-08-28T00:00:00Z","timestamp":1630108800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007195","name":"Universit\u00e0 degli Studi di Napoli Federico II","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007195","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Rapid Comm Mass Spectrometry"],"published-print":{"date-parts":[[2021,10,30]]},"abstract":"<jats:sec>\n                    <jats:title>Rationale<\/jats:title>\n                    <jats:p>\n                      Inflammation is a cascade of events mediated by a cytokine network triggering the cellular response. In order to monitor the modulation of the crucial inflammatory proteins, e.g., Tumour Necrosis Factor\u2010\u03b1 (TNF\u2010\u03b1), Interferon\u2010\u03b3 (INF\u2010\u03b3), Interleukin\u20108 (IL\u20108) and Interleukin\u201010 (IL\u201010), upon stimulation with endotoxins, differentiated and undifferentiated THP\u20101 cells were treated with lipopolysaccharides (LPSs) from\n                      <jats:styled-content style=\"fixed-case\">\n                        <jats:italic>E. coli<\/jats:italic>\n                      <\/jats:styled-content>\n                      , key cell wall components of Gram\u2010negative bacteria.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>The multiple reaction monitoring mass spectrometry (MRM\u2010MS) method was optimized by using the standard proteins to be quantified, in order to construct external calibration curves and define the analytical parameters. The developed method was used to quantify the above\u2010mentioned inflammatory proteins in THP\u20101 differentiated cells upon stimulation with LPSs with high accuracy, sensitivity, and robustness.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The analysis of such proteins in MRM mode allowed the kinetics of stimulation along the time up to 24\u2009h to be followed and the MS results were found to be comparable with those obtained by Western\u2010blotting. A significant increase in TNF\u2010\u03b1 release triggered a cascade mechanism leading to the production of INF\u2010\u03b3 and IL\u20108. IL\u201010, instead, was found to be constant throughout the process.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      The developed MRM\u2010MS method allowed the quantification of TNF\u2010\u03b1, INF\u2010\u03b3, IL\u20108 and IL\u201010 along a time\u2010course from 2 to 24\u2009h. Hence, a trace of the kinetics of the inflammatory response in THP\u20101 cells upon stimulation with\n                      <jats:styled-content style=\"fixed-case\">\n                        <jats:italic>E. coli<\/jats:italic>\n                      <\/jats:styled-content>\n                      LPSs was obtained. Finally, the extensibility of the developed MRM method to serum samples and other matrices demonstrated the versatility of the approach and the possibility to quantify multiple target proteins in different biological samples by using a few microliters in a single analysis.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/rcm.9166","type":"journal-article","created":{"date-parts":[[2021,7,16]],"date-time":"2021-07-16T12:56:31Z","timestamp":1626440191000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Inflammation protein quantification by multiple reaction monitoring mass spectrometry in lipopolysaccharide\u2010stimulated THP\u20101 cells"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1491-8966","authenticated-orcid":false,"given":"Anna","family":"Illiano","sequence":"first","affiliation":[{"name":"CEINGE Advanced Biotechnologies University of Naples Federico II  Naples Italy"},{"name":"Department of Chemical Sciences University of Naples Federico II  Naples Italy"},{"name":"Consorzio Interuniversitario Istituto Nazionale Biostrutture e Biosistemi  Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9169-3452","authenticated-orcid":false,"given":"Gabriella","family":"Pinto","sequence":"additional","affiliation":[{"name":"Department of Chemical Sciences University of Naples Federico II  Naples Italy"},{"name":"Consorzio Interuniversitario Istituto Nazionale Biostrutture e Biosistemi  Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2391-0237","authenticated-orcid":false,"given":"Rosa","family":"Gaglione","sequence":"additional","affiliation":[{"name":"Department of Chemical Sciences University of Naples Federico II  Naples 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