Transcriptomic profiling of the development of the inflammatory response in human monocytes in vitro
Monocytes/macrophages are key players in all phases of physiological and pathological inflammation. To understanding the regulation of macrophage functional differentiation during inflammation, we designed an in vitro model that recapitulates the different phases of the reaction (recruitment, initia...
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| Veröffentlicht in: | PloS one 2014-02, Vol.9 (2), p.e87680-e87680 |
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| description | Monocytes/macrophages are key players in all phases of physiological and pathological inflammation. To understanding the regulation of macrophage functional differentiation during inflammation, we designed an in vitro model that recapitulates the different phases of the reaction (recruitment, initiation, development, and resolution), based on human primary blood monocytes exposed to sequential changes in microenvironmental conditions. All reaction phases were profiled by transcriptomic microarray analysis. Distinct clusters of genes were identified that are differentially regulated through the different phases of inflammation. The gene sets defined by GSEA analysis revealed that the inflammatory phase was enriched in inflammatory pathways, while the resolution phase comprised pathways related to metabolism and gene rearrangement. By comparing gene clusters differentially expressed in monocytes vs. M1 and vs. M2 macrophages extracted from an in-house created meta-database, it was shown that cells in the model resemble M1 during the inflammatory phase and M2 during resolution. The validation of inflammatory and transcriptional factors by qPCR and ELISA confirmed the transcriptomic profiles in the different phases of inflammation. The accurate description of the development of the human inflammatory reaction provided by this in vitro kinetic model can help in identifying regulatory mechanisms in physiological conditions and during pathological derangements. |
| doi_str_mv | 10.1371/journal.pone.0087680 |
| format | Article |
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To understanding the regulation of macrophage functional differentiation during inflammation, we designed an in vitro model that recapitulates the different phases of the reaction (recruitment, initiation, development, and resolution), based on human primary blood monocytes exposed to sequential changes in microenvironmental conditions. All reaction phases were profiled by transcriptomic microarray analysis. Distinct clusters of genes were identified that are differentially regulated through the different phases of inflammation. The gene sets defined by GSEA analysis revealed that the inflammatory phase was enriched in inflammatory pathways, while the resolution phase comprised pathways related to metabolism and gene rearrangement. By comparing gene clusters differentially expressed in monocytes vs. M1 and vs. M2 macrophages extracted from an in-house created meta-database, it was shown that cells in the model resemble M1 during the inflammatory phase and M2 during resolution. The validation of inflammatory and transcriptional factors by qPCR and ELISA confirmed the transcriptomic profiles in the different phases of inflammation. The accurate description of the development of the human inflammatory reaction provided by this in vitro kinetic model can help in identifying regulatory mechanisms in physiological conditions and during pathological derangements.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0087680</identifier><identifier>PMID: 24498352</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adult ; Biology ; Blood & organ donations ; Cell Differentiation - genetics ; Cells, Cultured ; Cellular Microenvironment - genetics ; Chemokine CCL5 - genetics ; Chemokine CCL5 - metabolism ; Cluster Analysis ; Councils ; Cytokines ; DNA microarrays ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene clusters ; Gene expression ; Gene Expression Profiling ; Gene rearrangement ; Genes ; Homeostasis ; Human behavior ; Humans ; Immune system ; Inflammation ; Inflammation - genetics ; Inflammation - metabolism ; Inflammatory response ; Interleukin-6 - genetics ; Interleukin-6 - metabolism ; Interleukin-8 - genetics ; Interleukin-8 - metabolism ; Life sciences ; Macrophages ; Macrophages - cytology ; Macrophages - metabolism ; Male ; Metabolism ; Middle Aged ; Monocytes ; Monocytes - cytology ; Monocytes - metabolism ; Oligonucleotide Array Sequence Analysis ; Pathways ; Phases ; Physiology ; Recruitment ; Regulatory mechanisms (biology) ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction - genetics ; Studies ; Transcription factors ; Young Adult</subject><ispartof>PloS one, 2014-02, Vol.9 (2), p.e87680-e87680</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Italiani et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Italiani et al 2014 Italiani et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-9691855f2b08c4b6eee3ffeea834ffb0400d239e40af74192aaf8c898e210cc43</citedby><cites>FETCH-LOGICAL-c758t-9691855f2b08c4b6eee3ffeea834ffb0400d239e40af74192aaf8c898e210cc43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3912012/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3912012/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,729,782,786,866,887,2106,2932,23875,27933,27934,53800,53802</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24498352$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Poli, Guido</contributor><creatorcontrib>Italiani, Paola</creatorcontrib><creatorcontrib>Mazza, Emilia M C</creatorcontrib><creatorcontrib>Lucchesi, Davide</creatorcontrib><creatorcontrib>Cifola, Ingrid</creatorcontrib><creatorcontrib>Gemelli, Claudia</creatorcontrib><creatorcontrib>Grande, Alexis</creatorcontrib><creatorcontrib>Battaglia, Cristina</creatorcontrib><creatorcontrib>Bicciato, Silvio</creatorcontrib><creatorcontrib>Boraschi, Diana</creatorcontrib><title>Transcriptomic profiling of the development of the inflammatory response in human monocytes in vitro</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Monocytes/macrophages are key players in all phases of physiological and pathological inflammation. To understanding the regulation of macrophage functional differentiation during inflammation, we designed an in vitro model that recapitulates the different phases of the reaction (recruitment, initiation, development, and resolution), based on human primary blood monocytes exposed to sequential changes in microenvironmental conditions. All reaction phases were profiled by transcriptomic microarray analysis. Distinct clusters of genes were identified that are differentially regulated through the different phases of inflammation. The gene sets defined by GSEA analysis revealed that the inflammatory phase was enriched in inflammatory pathways, while the resolution phase comprised pathways related to metabolism and gene rearrangement. By comparing gene clusters differentially expressed in monocytes vs. M1 and vs. M2 macrophages extracted from an in-house created meta-database, it was shown that cells in the model resemble M1 during the inflammatory phase and M2 during resolution. The validation of inflammatory and transcriptional factors by qPCR and ELISA confirmed the transcriptomic profiles in the different phases of inflammation. The accurate description of the development of the human inflammatory reaction provided by this in vitro kinetic model can help in identifying regulatory mechanisms in physiological conditions and during pathological derangements.</description><subject>Adult</subject><subject>Biology</subject><subject>Blood & organ donations</subject><subject>Cell Differentiation - genetics</subject><subject>Cells, Cultured</subject><subject>Cellular Microenvironment - genetics</subject><subject>Chemokine CCL5 - genetics</subject><subject>Chemokine CCL5 - metabolism</subject><subject>Cluster Analysis</subject><subject>Councils</subject><subject>Cytokines</subject><subject>DNA microarrays</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Female</subject><subject>Gene clusters</subject><subject>Gene expression</subject><subject>Gene Expression Profiling</subject><subject>Gene rearrangement</subject><subject>Genes</subject><subject>Homeostasis</subject><subject>Human behavior</subject><subject>Humans</subject><subject>Immune system</subject><subject>Inflammation</subject><subject>Inflammation - 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To understanding the regulation of macrophage functional differentiation during inflammation, we designed an in vitro model that recapitulates the different phases of the reaction (recruitment, initiation, development, and resolution), based on human primary blood monocytes exposed to sequential changes in microenvironmental conditions. All reaction phases were profiled by transcriptomic microarray analysis. Distinct clusters of genes were identified that are differentially regulated through the different phases of inflammation. The gene sets defined by GSEA analysis revealed that the inflammatory phase was enriched in inflammatory pathways, while the resolution phase comprised pathways related to metabolism and gene rearrangement. By comparing gene clusters differentially expressed in monocytes vs. M1 and vs. M2 macrophages extracted from an in-house created meta-database, it was shown that cells in the model resemble M1 during the inflammatory phase and M2 during resolution. The validation of inflammatory and transcriptional factors by qPCR and ELISA confirmed the transcriptomic profiles in the different phases of inflammation. The accurate description of the development of the human inflammatory reaction provided by this in vitro kinetic model can help in identifying regulatory mechanisms in physiological conditions and during pathological derangements.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24498352</pmid><doi>10.1371/journal.pone.0087680</doi><tpages>e87680</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Adult Biology Blood & organ donations Cell Differentiation - genetics Cells, Cultured Cellular Microenvironment - genetics Chemokine CCL5 - genetics Chemokine CCL5 - metabolism Cluster Analysis Councils Cytokines DNA microarrays Enzyme-Linked Immunosorbent Assay Female Gene clusters Gene expression Gene Expression Profiling Gene rearrangement Genes Homeostasis Human behavior Humans Immune system Inflammation Inflammation - genetics Inflammation - metabolism Inflammatory response Interleukin-6 - genetics Interleukin-6 - metabolism Interleukin-8 - genetics Interleukin-8 - metabolism Life sciences Macrophages Macrophages - cytology Macrophages - metabolism Male Metabolism Middle Aged Monocytes Monocytes - cytology Monocytes - metabolism Oligonucleotide Array Sequence Analysis Pathways Phases Physiology Recruitment Regulatory mechanisms (biology) Reverse Transcriptase Polymerase Chain Reaction Signal Transduction - genetics Studies Transcription factors Young Adult |
| title | Transcriptomic profiling of the development of the inflammatory response in human monocytes in vitro |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-02T18%3A48%3A44IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Transcriptomic%20profiling%20of%20the%20development%20of%20the%20inflammatory%20response%20in%20human%20monocytes%20in%20vitro&rft.jtitle=PloS%20one&rft.au=Italiani,%20Paola&rft.date=2014-02-03&rft.volume=9&rft.issue=2&rft.spage=e87680&rft.epage=e87680&rft.pages=e87680-e87680&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0087680&rft_dat=%3Cgale_plos_%3EA478831918%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1494057633&rft_id=info:pmid/24498352&rft_galeid=A478831918&rft_doaj_id=oai_doaj_org_article_ef8c976d1ec14ab68b93585bb725bc99&rfr_iscdi=true |