{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T21:11:32Z","timestamp":1789938692533,"version":"4.0.1"},"reference-count":20,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,5]],"date-time":"2021-07-05T00:00:00Z","timestamp":1625443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>The development of advanced single-cell technologies to decipher inter-cellular heterogeneity has enabled the dynamic assessment of individual cells behavior over time, overcoming the limitation of traditional assays. Here, we evaluated the feasibility of an advanced microfluidic assay combined to fluorescence microscopy to address the behavior of circulating monocytes from septic shock patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Seven septic shock patients and ten healthy volunteers were enrolled in the study. Using the proposed microfluidic assay we investigated the production over time of LPS-elicited TNF\u03b1 by single monocytes encapsulated within droplets. Cellular endocytic activity was assessed by internalization of magnetic nanoparticles. Besides, we assessed HLA-DR membrane expression and LPS-induced TNF\u03b1 production in monocytes through classical flow cytometry assays.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Consistent with the flow cytometry results, the total number of TNF\u03b1 molecules secreted by encapsulated single monocytes was significantly decreased in septic shock patients compared to healthy donors. TNF\u03b1 production was dampened as soon as 30 and 60 minutes after LPS stimulation in monocytes from septic patients. Furthermore, the microfluidic assay revealed heterogeneous individual behavior of monocytes from septic shock patients. Of note, monocytes from both healthy donors and patients exhibited similar phagocytic activities over time.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The microfluidic assay highlights the functional heterogeneity of monocytes, and provides in-depth resolution in assessing the hallmark monocyte deactivation encountered in post-septic immunosuppression.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2021.686111","type":"journal-article","created":{"date-parts":[[2021,7,5]],"date-time":"2021-07-05T14:47:50Z","timestamp":1625496470000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Assessing the Functional Heterogeneity of Monocytes in Human Septic Shock: a Proof-of-Concept Microfluidic Assay of TNF\u03b1 Secretion"],"prefix":"10.3389","volume":"12","author":[{"given":"Jean-Fran\u00e7ois","family":"Llitjos","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yacine","family":"Bounab","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christophe","family":"Rousseau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Dixneuf","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Blandine","family":"Rimbault","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Daniel","family":"Chiche","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julien","family":"Textoris","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fr\u00e9d\u00e9ric","family":"P\u00e8ne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christophe","family":"V\u00e9drine","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1001\/jama.2016.0287","article-title":"The Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3)","volume":"315","author":"Singer","year":"2016","journal-title":"JAMA"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1007\/s00134-006-0204-8","article-title":"Persisting Low Monocyte Human Leukocyte Antigen-DR Expression Predicts Mortality in Septic Shock","volume":"32","author":"Monneret","year":"2006","journal-title":"Intensive Care Med"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1038\/nri3552","article-title":"Sepsis-Induced Immunosuppression: From Cellular Dysfunctions to Immunotherapy","volume":"13","author":"Hotchkiss","year":"2013","journal-title":"Nat Rev Immunol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1007\/s00134-010-1962-x","article-title":"Low Monocyte Human Leukocyte Antigen-DR Is Independently Associated With Nosocomial Infections After Septic Shock","volume":"36","author":"Landelle","year":"2010","journal-title":"Intensive Care Med"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.08.027","article-title":"Pathogen Cell-To-Cell Variability Drives Heterogeneity in Host Immune Responses","volume":"162","author":"Avraham","year":"2015","journal-title":"Cell"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1038\/nature12172","article-title":"Single-Cell Transcriptomics Reveals Bimodality in Expression and Splicing in Immune Cells","volume":"498","author":"Shalek","year":"2013","journal-title":"Nature"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-020-0354-0","article-title":"Dynamic Single-Cell Phenotyping of Immune Cells Using the Microfluidic Platform DropMap","volume":"15","author":"Bounab","year":"2020","journal-title":"Nat Protoc"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1097\/00003246-199811000-00016","article-title":"Use of the SOFA Score to Assess the Incidence of Organ Dysfunction\/Failure in Intensive Care Units: Results of a Multicenter, Prospective Study. 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