{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T19:57:01Z","timestamp":1786737421236,"version":"build-2736575974"},"reference-count":40,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"5","license":[{"start":{"date-parts":[[2020,3,3]],"date-time":"2020-03-03T00:00:00Z","timestamp":1583193600000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Hepatol Commun"],"published-print":{"date-parts":[[2020,5]]},"abstract":"<jats:p>Intrahepatic macrophages influence the composition of the microenvironment, host immune response to liver injury, and development of fibrosis. Compared with stellate cells, the role of macrophages in the development of fibrosis remains unclear. Multispectral imaging allows detection of multiple markers <jats:italic toggle=\"yes\">in situ<\/jats:italic> in human formalin\u2010fixed, paraffin\u2010embedded tissue. This cutting\u2010edge technology is ideal for analyzing human liver tissues, as it allows spectral unmixing of fluorophore signals, subtraction of auto\u2010fluorescence, and preservation of hepatic architecture. We analyzed five different antibodies commonly observed on macrophage populations (CD68, MAC387, CD163, CD14, and CD16). After optimization of the monoplex stains and development of a Spectral Library, we combined all of the antibodies into a multiplex protocol and used them to stain biopsies collected from representative patients with chronic liver diseases, including chronic hepatitis C, nonalcoholic steatohepatitis, and autoimmune hepatitis. Various imaging modalities were tested, including cell phenotyping, tissue segmentation, t\u2010distributed stochastic neighbor embedding plots, and phenotype matrices that facilitated comparison and visualization of the identified macrophage and other cellular profiles. We then tested the feasibility of this platform to analyze numerous regions of interest from liver biopsies with multiple patients per group, using batch analysis algorithms. Five populations showed significant differences between patients positive for hepatitis C virus with advanced fibrosis when compared with controls. Three of these were significantly increased in patients with advanced fibrosis when compared to controls, and these included CD163+CD16+, CD68+, and CD68+MAC387+. <jats:italic toggle=\"yes\">Conclusion:<\/jats:italic> Spectral imaging microscopy is a powerful tool that enables <jats:italic toggle=\"yes\">in situ<\/jats:italic> analysis of macrophages and other cells in human liver biopsies and may lead to more personalized therapeutic approaches in the future.<\/jats:p>","DOI":"10.1002\/hep4.1494","type":"journal-article","created":{"date-parts":[[2020,3,4]],"date-time":"2020-03-04T07:33:16Z","timestamp":1583307196000},"page":"708-723","source":"Crossref","is-referenced-by-count":22,"title":["Multispectral Imaging Enables Characterization of Intrahepatic Macrophages in Patients With Chronic Liver Disease"],"prefix":"10.1097","volume":"4","author":[{"given":"Omar A.","family":"Saldarriaga","sequence":"first","affiliation":[{"name":"Department of Pathology,University of Texas Medical Branch,Galveston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin","family":"Freiberg","sequence":"additional","affiliation":[{"name":"Visiopharm,Hoersholm,Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Santhoshi","family":"Krishnan","sequence":"additional","affiliation":[{"name":"Department of Electrical and Computer Engineering,Rice University,Houston,TX"},{"name":"Department of Computational Medicine and Bioinformatics,University of Michigan,Ann Arbor,MI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arvind","family":"Rao","sequence":"additional","affiliation":[{"name":"Department of Electrical and Computer Engineering,Rice University,Houston,TX"},{"name":"Department of Computational Medicine and Bioinformatics,University of Michigan,Ann Arbor,MI"},{"name":"Department of Radiation Oncology,University of Michigan,Ann Arbor,MI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jared","family":"Burks","sequence":"additional","affiliation":[{"name":"Department of Leukemia,University of Texas MD Anderson Cancer Center,Houston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1231-4117","authenticated-orcid":false,"given":"Adam L.","family":"Booth","sequence":"additional","affiliation":[{"name":"Department of Pathology,University of Texas Medical Branch,Galveston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bradley","family":"Dye","sequence":"additional","affiliation":[{"name":"Department of Pathology,University of Texas Medical Branch,Galveston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Netanya","family":"Utay","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine,University of Texas Health Science Center at Houston,Houston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monique","family":"Ferguson","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine,University of Texas Medical Branch,Galveston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdellah","family":"Akil","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology,University of Texas Medical Branch,Galveston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minkyung","family":"Yi","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology,University of Texas Medical Branch,Galveston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2054-684X","authenticated-orcid":false,"given":"Laura","family":"Beretta","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Oncology,University of Texas MD Anderson Cancer Center,Houston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0645-7621","authenticated-orcid":false,"given":"Heather L.","family":"Stevenson","sequence":"additional","affiliation":[{"name":"Department of Pathology,University of Texas Medical Branch,Galveston,TX"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","published-online":{"date-parts":[[2020,3,3]]},"reference":[{"key":"hep41494-bib-0001-20240114","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1016\/j.molimm.2017.02.018","article-title":"The role of Kupffer cells in hepatic diseases","volume":"85","year":"2017","journal-title":"Mol Immunol"},{"key":"hep41494-bib-0002-20240114","first-page":"763","article-title":"The study of liver macrophages polarization in patients with autoimmune hepatitis","volume":"56","year":"2017","journal-title":"Zhonghua Nei Ke Za Zhi"},{"key":"hep41494-bib-0003-20240114","doi-asserted-by":"crossref","first-page":"3874","DOI":"10.3892\/mmr.2016.4998","article-title":"Altered function of monocytes\/macrophages in patients with autoimmune hepatitis","volume":"13","year":"2016","journal-title":"Mol Med Rep"},{"key":"hep41494-bib-0004-20240114","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/S0171-2985(81)80090-3","article-title":"Influence of various detachment procedures on the functional state of cultured murine macrophages","volume":"159","year":"1981","journal-title":"Immunobiology"},{"key":"hep41494-bib-0005-20240114","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/j.jim.2015.08.001","article-title":"Impact of detachment methods on M2 macrophage phenotype and function","volume":"426","year":"2015","journal-title":"J Immunol Methods"},{"key":"hep41494-bib-0006-20240114","doi-asserted-by":"crossref","first-page":"347","DOI":"10.4049\/jimmunol.1701262","article-title":"Eight\u2010color multiplex immunohistochemistry for simultaneous detection of multiple immune checkpoint molecules within the tumor microenvironment","volume":"200","year":"2018","journal-title":"J Immunol"},{"key":"hep41494-bib-0007-20240114","doi-asserted-by":"crossref","first-page":"4383","DOI":"10.1038\/s41467-018-06318-7","article-title":"Single cell RNA sequencing of human liver reveals distinct intrahepatic macrophage populations","volume":"9","year":"2018","journal-title":"Nat Commun"},{"key":"hep41494-bib-0008-20240114","article-title":"Cytometry TOF identifies alveolar macrophage subtypes in acute respiratory distress syndrome","volume":"3","year":"2018","journal-title":"JCI Insight"},{"key":"hep41494-bib-0009-20240114","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s40169-015-0047-4","article-title":"The significance of macrophage polarization subtypes for animal models of tissue fibrosis and human fibrotic diseases","volume":"4","year":"2015","journal-title":"Clin Transl Med"},{"key":"hep41494-bib-0010-20240114","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1002\/path.5026","article-title":"Multispectral imaging for quantitative and compartment\u2010specific immune infiltrates reveals distinct immune profiles that classify lung cancer patients","volume":"244","year":"2018","journal-title":"J Pathol"},{"key":"hep41494-bib-0011-20240114","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.humpath.2012.05.009","article-title":"A colorful future of quantitative pathology: validation of Vectra technology using chromogenic multiplexed immunohistochemistry and prostate tissue microarrays","volume":"44","year":"2013","journal-title":"Hum Pathol"},{"key":"hep41494-bib-0012-20240114","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1369\/0022155411398488","article-title":"Factors influencing the degradation of archival formalin\u2010fixed paraffin\u2010embedded tissue sections","volume":"59","year":"2011","journal-title":"J Histochem Cytochem"},{"key":"hep41494-bib-0013-20240114","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.jhep.2014.02.031","article-title":"Image analysis of liver biopsy samples measures fibrosis and predicts clinical outcome","volume":"61","year":"2014","journal-title":"J Hepatol"},{"key":"hep41494-bib-0014-20240114","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.ymeth.2014.08.016","article-title":"Multiplexed immunohistochemistry, imaging, and quantitation: a review, with an assessment of Tyramide signal amplification, multispectral imaging and multiplex analysis","volume":"70","year":"2014","journal-title":"Methods"},{"key":"hep41494-bib-0015-20240114","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1007\/978-1-4939-2742-5_16","article-title":"Tyramide signal amplification for immunofluorescent enhancement","volume":"1318","year":"2015","journal-title":"Methods Mol Biol"},{"key":"hep41494-bib-0016-20240114","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1136\/gut.46.2.260","article-title":"Increases in intrahepatic CD68 positive cells, MAC387 positive cells, and proinflammatory cytokines (particularly interleukin 18) in chronic hepatitis C infection","volume":"46","year":"2000","journal-title":"Gut"},{"key":"hep41494-bib-0017-20240114","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1002\/hep.26016","article-title":"Monocyte subsets in human liver disease show distinct phenotypic and functional characteristics","volume":"57","year":"2013","journal-title":"Hepatology"},{"key":"hep41494-bib-0018-20240114","doi-asserted-by":"crossref","first-page":"44356","DOI":"10.1038\/srep44356","article-title":"Immunofluorescence identifies distinct subsets of endothelial cells in the human liver","volume":"7","year":"2017","journal-title":"Sci Rep"},{"key":"hep41494-bib-0019-20240114","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1189\/jlb.0604332","article-title":"Development and functional consequences of LPS tolerance in sinusoidal endothelial cells of the liver","volume":"77","year":"2005","journal-title":"J Leukoc Biol"},{"key":"hep41494-bib-0020-20240114","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1016\/0168-8278(95)80226-6","article-title":"Histological grading and staging of chronic hepatitis","volume":"22","year":"1995","journal-title":"J Hepatol"},{"key":"hep41494-bib-0021-20240114","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1038\/cmi.2015.104","article-title":"Hepatic macrophages in homeostasis and liver diseases: from pathogenesis to novel therapeutic strategies","volume":"13","year":"2016","journal-title":"Cell Mol Immunol"},{"key":"hep41494-bib-0022-20240114","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1038\/nri.2017.11","article-title":"Liver macrophages in tissue homeostasis and disease","volume":"17","year":"2017","journal-title":"Nat Rev Immunol"},{"key":"hep41494-bib-0023-20240114","doi-asserted-by":"crossref","first-page":"1754","DOI":"10.1002\/hep.29477","article-title":"A randomized, placebo\u2010controlled trial of cenicriviroc for treatment of nonalcoholic steatohepatitis with fibrosis","volume":"67","year":"2018","journal-title":"Hepatology"},{"key":"hep41494-bib-0024-20240114","article-title":"Kupffer cells undergo fundamental changes during the development of experimental NASH and are critical in initiating liver damage and inflammation","volume":"11","year":"2016","journal-title":"PLoS One"},{"key":"hep41494-bib-0025-20240114","first-page":"25","article-title":"Increase of infiltrating monocytes in the livers of patients with chronic liver diseases","volume":"21","year":"2016","journal-title":"Discov Med"},{"key":"hep41494-bib-0026-20240114","doi-asserted-by":"crossref","first-page":"39520","DOI":"10.1038\/srep39520","article-title":"Chronic hepatitis C infection\u2010induced liver fibrogenesis is associated with M2 macrophage activation","volume":"6","year":"2016","journal-title":"Sci Rep"},{"key":"hep41494-bib-0027-20240114","doi-asserted-by":"crossref","first-page":"893","DOI":"10.3389\/fimmu.2019.00893","article-title":"The role of myeloid\u2010derived cells in the progression of liver disease","volume":"10","year":"2019","journal-title":"Front Immunol"},{"key":"hep41494-bib-0028-20240114","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1038\/nrgastro.2015.200","article-title":"Immunology in the liver\u2014from homeostasis to disease","volume":"13","year":"2016","journal-title":"Nat Rev Gastroenterol Hepatol"},{"key":"hep41494-bib-0029-20240114","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.jcmgh.2015.12.005","article-title":"Hepatitis C virus\u2010induced monocyte differentiation into polarized M2 macrophages promotes stellate cell activation via TGF\u2010beta","volume":"2","year":"2016","journal-title":"Cell Mol Gastroenterol Hepatol"},{"key":"hep41494-bib-0030-20240114","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1111\/liv.12050","article-title":"Portal, but not lobular, macrophages express matrix metalloproteinase\u20109: association with the ductular reaction and fibrosis in chronic hepatitis C","volume":"33","year":"2013","journal-title":"Liver Int"},{"key":"hep41494-bib-0031-20240114","article-title":"Functional contribution of elevated circulating and hepatic non\u2010classical CD14CD16 monocytes to inflammation and human liver fibrosis","volume":"5","year":"2010","journal-title":"PLoS One"},{"key":"hep41494-bib-0032-20240114","article-title":"Differential expression of CD163 on monocyte subsets in healthy and HIV\u20101 infected individuals","volume":"6","year":"2011","journal-title":"PLoS One"},{"key":"hep41494-bib-0033-20240114","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1038\/s41598-018-37071-y","article-title":"Fibrogenic gene expression in hepatic stellate cells induced by HCV and HIV replication in a three cell co\u2010culture model system","volume":"9","year":"2019","journal-title":"Sci Rep"},{"key":"hep41494-bib-0034-20240114","first-page":"490","article-title":"Intrahepatic CD206","volume":"67","year":"2017","journal-title":"J Hepatol"},{"key":"hep41494-bib-0035-20240114","doi-asserted-by":"crossref","first-page":"2948","DOI":"10.3389\/fimmu.2018.02948","article-title":"The role of monocytes and macrophages in acute and acute\u2010on\u2010chronic liver failure","volume":"9","year":"2018","journal-title":"Front Immunol"},{"key":"hep41494-bib-0036-20240114","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.1016\/j.jhep.2017.02.026","article-title":"Targeting hepatic macrophages to treat liver diseases","volume":"66","year":"2017","journal-title":"J Hepatol"},{"key":"hep41494-bib-0037-20240114","doi-asserted-by":"crossref","first-page":"1270","DOI":"10.1002\/hep.29544","article-title":"Therapeutic inhibition of inflammatory monocyte recruitment reduces steatohepatitis and liver fibrosis","volume":"67","year":"2018","journal-title":"Hepatology"},{"key":"hep41494-bib-0038-20240114","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1056\/NEJMoa1604958","article-title":"Mutations associated with acquired resistance to PD\u20101 blockade in melanoma","volume":"375","year":"2016","journal-title":"N Engl J Med"},{"key":"hep41494-bib-0039-20240114","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1158\/2326-6066.CIR-14-0244","article-title":"Resistance to antiangiogenic therapy is associated with an immunosuppressive tumor microenvironment in metastatic renal cell carcinoma","volume":"3","year":"2015","journal-title":"Cancer Immunol Res"},{"key":"hep41494-bib-0040-20240114","first-page":"327ra326","article-title":"The tumor microenvironment and Immunoscore are critical determinants of dissemination to distant metastasis","volume":"8","year":"2016","journal-title":"Sci Transl Med"}],"container-title":["Hepatology Communications"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fhep4.1494","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/hep4.1494","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/hep4.1494","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.lww.com\/10.1002\/hep4.1494","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,14]],"date-time":"2024-01-14T17:06:54Z","timestamp":1705252014000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.lww.com\/10.1002\/hep4.1494"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,3]]},"references-count":40,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2020]]}},"URL":"https:\/\/doi.org\/10.1002\/hep4.1494","relation":{},"ISSN":["2471-254X","2471-254X"],"issn-type":[{"value":"2471-254X","type":"print"},{"value":"2471-254X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,3]]}}}