{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,22]],"date-time":"2026-01-22T23:34:00Z","timestamp":1769124840527,"version":"3.49.0"},"posted":{"date-parts":[[2023,4,3]]},"group-title":"Immunology","reference-count":68,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,9,30]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  <jats:italic>Bacillus anthracis<\/jats:italic>\n                  peptidoglycan (PGN) is a major component of the bacterial cell wall and a key pathogen-associated molecular pattern (PAMP) contributing to anthrax pathology, including organ dysfunction and coagulopathy. Increases in apoptotic lymphocytes are a late-stage feature of anthrax and sepsis, suggesting there is a defect in apoptotic clearance. Here, we tested the hypothesis that\n                  <jats:italic>B. anthracis<\/jats:italic>\n                  PGN inhibits the capacity of human monocyte-derived macrophages (M\u03a6) to efferocytose apoptotic cells. Exposure of CD163\n                  <jats:sup>+<\/jats:sup>\n                  CD206\n                  <jats:sup>+<\/jats:sup>\n                  M\u03a6 to PGN for 24h impaired efferocytosis in a manner dependent on human serum opsonins but independent of complement component C3. PGN treatment reduced cell surface expression of the pro-efferocytic signaling receptors MERTK, TYRO3, AXL, integrin \u03b1V\u03b25, CD36 and TIM-3, whereas TIM-1, \u03b1V\u03b23, CD300b, CD300f, STABILIN-1 and STABILIN-2 were unaffected. ADAM17 is a major membrane-bound protease implicated in mediating efferocytotic receptor cleavage. We found multiple ADAM17-mediated substrates increased in PGN-treated supernatant suggesting involvement of membrane-bound proteases. ADAM17 inhibitors TAPI-0 and Marimastat prevented TNF release, indicating effective protease inhibition, and modestly increased cell-surface levels of MerTK and TIM-3 but only partially restored efferocytic capacity by PGN-treated M\u03a6. We conclude that human serum factors are required for optimal recognition of PGN by human M\u03a6 and that\n                  <jats:italic>B. anthracis<\/jats:italic>\n                  PGN inhibits efferocytosis in part by reducing cell surface expression of MERTK and TIM-3.\n                <\/jats:p>","DOI":"10.1101\/2023.03.30.535001","type":"posted-content","created":{"date-parts":[[2023,4,3]],"date-time":"2023-04-03T17:35:17Z","timestamp":1680543317000},"source":"Crossref","is-referenced-by-count":1,"title":["Peptidoglycan from\n                  <i>Bacillus anthracis Inhibits<\/i>\n                  Human Macrophage Efferocytosis in Part by Reducing Cell Surface Expression of MERTK and TIM-3"],"prefix":"10.64898","author":[{"given":"Joshua S","family":"Mytych","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zijian","family":"Pan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Charmaine","family":"Lopez-Davis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nancy","family":"Redinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christina","family":"Lawrence","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jadith","family":"Ziegler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4809-9885","authenticated-orcid":false,"given":"Narcis I.","family":"Popescu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Judith A.","family":"James","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3009-2017","authenticated-orcid":false,"given":"A. Darise","family":"Farris","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023100513550562000_2023.03.30.535001v3.1","doi-asserted-by":"publisher","DOI":"10.12998\/wjcc.v3.i1.20"},{"key":"2023100513550562000_2023.03.30.535001v3.2","doi-asserted-by":"crossref","unstructured":"Rubinson, L.M.P. , A. Corey , and D. Hanfling , Estimation of Time Period for Effective Human Inhalational Anthrax Treatment Including Antitoxin Therapy. PLoS Curr, 2017. 9.","DOI":"10.1371\/currents.outbreaks.7896c43f69838f17ce1c2c372e79d55d"},{"key":"2023100513550562000_2023.03.30.535001v3.3","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(19)32989-7"},{"key":"2023100513550562000_2023.03.30.535001v3.4","doi-asserted-by":"crossref","first-page":"1918","DOI":"10.3389\/fimmu.2020.01918","article-title":"The Critical Roles and Mechanisms of Immune Cell Death in Sepsis","volume":"11","year":"2020","journal-title":"Front Immunol"},{"key":"2023100513550562000_2023.03.30.535001v3.5","doi-asserted-by":"crossref","first-page":"650184","DOI":"10.3389\/fimmu.2021.650184","article-title":"Circulating Histones in Sepsis: Potential Outcome Predictors and Therapeutic Targets","volume":"12","year":"2021","journal-title":"Front Immunol"},{"key":"2023100513550562000_2023.03.30.535001v3.6","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2053"},{"key":"2023100513550562000_2023.03.30.535001v3.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijid.2012.03.007"},{"key":"2023100513550562000_2023.03.30.535001v3.8","doi-asserted-by":"publisher","DOI":"10.1097\/01.CCM.0000074719.40109.95"},{"key":"2023100513550562000_2023.03.30.535001v3.9","doi-asserted-by":"publisher","DOI":"10.1038\/nature20138"},{"key":"2023100513550562000_2023.03.30.535001v3.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2017.07.019"},{"issue":"4","key":"2023100513550562000_2023.03.30.535001v3.11","doi-asserted-by":"crossref","first-page":"1536","DOI":"10.1016\/j.jaci.2018.10.046","article-title":"Transcriptional and functional diversity of human macrophage repolarization","volume":"143","year":"2019","journal-title":"J Allergy Clin Immunol"},{"key":"2023100513550562000_2023.03.30.535001v3.12","doi-asserted-by":"publisher","DOI":"10.1016\/J.IMMUNI.2019.04.018"},{"issue":"5","key":"2023100513550562000_2023.03.30.535001v3.13","doi-asserted-by":"crossref","first-page":"913","DOI":"10.1016\/j.immuni.2017.10.006","article-title":"Tissue-Resident Macrophages Are Locally Programmed for Silent Clearance of Apoptotic Cells","volume":"47","year":"2017","journal-title":"Immunity"},{"key":"2023100513550562000_2023.03.30.535001v3.14","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.2005.02934.x"},{"key":"2023100513550562000_2023.03.30.535001v3.15","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2012-04-423525"},{"key":"2023100513550562000_2023.03.30.535001v3.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2014.06.008"},{"key":"2023100513550562000_2023.03.30.535001v3.17","doi-asserted-by":"crossref","unstructured":"Roszer, T. , Understanding the Mysterious M2 Macrophage through Activation Markers and Effector Mechanisms. Mediators of Inflammation, 2015. 2015.","DOI":"10.1155\/2015\/816460"},{"key":"2023100513550562000_2023.03.30.535001v3.18","doi-asserted-by":"crossref","first-page":"2536","DOI":"10.3389\/fimmu.2019.02536","article-title":"DAMPs and NETs in Sepsis","volume":"10","year":"2019","journal-title":"Front Immunol"},{"issue":"8","key":"2023100513550562000_2023.03.30.535001v3.19","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1038\/s41573-022-00470-y","article-title":"Drugging the efferocytosis process: concepts and opportunities","volume":"21","year":"2022","journal-title":"Nat Rev Drug Discov"},{"key":"2023100513550562000_2023.03.30.535001v3.20","doi-asserted-by":"publisher","DOI":"10.1111\/imr.12453"},{"key":"2023100513550562000_2023.03.30.535001v3.21","doi-asserted-by":"crossref","unstructured":"Doran, A.C. , A. Yurdagul, Jr ., and I. Tabas , Efferocytosis in health and disease. Nat Rev Immunol, 2019.","DOI":"10.1038\/s41577-019-0240-6"},{"key":"2023100513550562000_2023.03.30.535001v3.22","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.arr.2019.01.007","article-title":"Effect of soluble cleavage products of important receptors\/ligands on efferocytosis: Their role in inflammatory, autoimmune and cardiovascular disease","volume":"50","year":"2019","journal-title":"Ageing Res Rev"},{"issue":"6","key":"2023100513550562000_2023.03.30.535001v3.23","doi-asserted-by":"crossref","first-page":"5133","DOI":"10.1007\/s11033-022-07224-4","article-title":"Potential diagnostic and prognostic of efferocytosis-related unwanted soluble receptors\/ligands as new non-invasive biomarkers in disorders: a review","volume":"49","year":"2022","journal-title":"Mol Biol Rep"},{"key":"2023100513550562000_2023.03.30.535001v3.24","doi-asserted-by":"crossref","unstructured":"Witas, R. , et al., Defective Efferocytosis in a Murine Model of Sjogren\u2019s Syndrome Is Mediated by Dysfunctional Mer Tyrosine Kinase Receptor. Int J Mol Sci, 2021. 22(18).","DOI":"10.3390\/ijms22189711"},{"key":"2023100513550562000_2023.03.30.535001v3.25","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1524292113"},{"key":"2023100513550562000_2023.03.30.535001v3.26","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0307193"},{"key":"2023100513550562000_2023.03.30.535001v3.27","doi-asserted-by":"crossref","unstructured":"Pan, Z. , et al., Bacillus anthracis Edema Toxin Inhibits Efferocytosis in Human Macrophages and Alters Efferocytic Receptor Signaling. Int J Mol Sci, 2019. 20(5).","DOI":"10.3390\/ijms20051167"},{"key":"2023100513550562000_2023.03.30.535001v3.28","doi-asserted-by":"crossref","unstructured":"Keshari, R.S. , et al., Complement C5 inhibition protects against hemolytic anemia and acute kidney injury in anthrax peptidoglycan-induced sepsis in baboons. Proc Natl Acad Sci U S A, 2021. 118(37).","DOI":"10.1073\/pnas.2104347118"},{"key":"2023100513550562000_2023.03.30.535001v3.29","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2017-10-813618"},{"key":"2023100513550562000_2023.03.30.535001v3.30","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.165.10.5392"},{"key":"2023100513550562000_2023.03.30.535001v3.31","doi-asserted-by":"publisher","DOI":"10.1038\/sj.embor.7400248"},{"key":"2023100513550562000_2023.03.30.535001v3.32","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M307198200"},{"issue":"2","key":"2023100513550562000_2023.03.30.535001v3.33","doi-asserted-by":"crossref","first-page":"e0193207","DOI":"10.1371\/journal.pone.0193207","article-title":"Neither Lys- and DAP-type peptidoglycans stimulate mouse or human innate immune cells via Toll-like receptor 2","volume":"13","year":"2018","journal-title":"PLoS One"},{"issue":"7927","key":"2023100513550562000_2023.03.30.535001v3.34","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1038\/s41586-022-05125-x","article-title":"Phosphorylation of muramyl peptides by NAGK is required for NOD2 activation","volume":"609","year":"2022","journal-title":"Nature"},{"key":"2023100513550562000_2023.03.30.535001v3.35","doi-asserted-by":"crossref","unstructured":"Girton, A.W. , et al., Serum Amyloid P and IgG Exhibit Differential Capabilities in the Activation of the Innate Immune System in Response to Bacillus anthracis Peptidoglycan. Infect Immun, 2018. 86(5).","DOI":"10.1128\/IAI.00076-18"},{"key":"2023100513550562000_2023.03.30.535001v3.36","doi-asserted-by":"crossref","unstructured":"Turner, S. , et al., Gram-Positive Bacteria Cell Wall Peptidoglycan Polymers Activate Human Dendritic Cells to Produce IL-23 and IL-1beta and Promote T(H)17 Cell Differentiation. Microorganisms, 2023. 11(1).","DOI":"10.3390\/microorganisms11010173"},{"key":"2023100513550562000_2023.03.30.535001v3.37","doi-asserted-by":"crossref","unstructured":"Popescu, N.I. , et al., C3 Opsonization of Anthrax Bacterium and Peptidoglycan Supports Recognition and Activation of Neutrophils. Microorganisms, 2020. 8(7).","DOI":"10.3390\/microorganisms8071039"},{"key":"2023100513550562000_2023.03.30.535001v3.38","doi-asserted-by":"crossref","unstructured":"Langer, M. , et al., Neither Lys- and DAP-type peptidoglycans stimulate mouse or human innate immune cells via Toll-like receptor 2. Plos One, 2018. 13(2).","DOI":"10.1371\/journal.pone.0193207"},{"key":"2023100513550562000_2023.03.30.535001v3.39","doi-asserted-by":"crossref","unstructured":"Moss, M.L. and D. Minond , Recent Advances in ADAM17 Research: A Promising Target for Cancer and Inflammation. Mediators Inflamm, 2017. 2017: p. 9673537.","DOI":"10.1155\/2017\/9673537"},{"key":"2023100513550562000_2023.03.30.535001v3.40","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.bcp.2019.02.033","article-title":"Inhibition of MMPs and ADAM\/ADAMTS","volume":"165","year":"2019","journal-title":"Biochem Pharmacol"},{"key":"2023100513550562000_2023.03.30.535001v3.41","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2015.01.012"},{"key":"2023100513550562000_2023.03.30.535001v3.42","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2016.05.011"},{"key":"2023100513550562000_2023.03.30.535001v3.43","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.263020"},{"key":"2023100513550562000_2023.03.30.535001v3.44","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.112.300683"},{"key":"2023100513550562000_2023.03.30.535001v3.45","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.488478"},{"issue":"2","key":"2023100513550562000_2023.03.30.535001v3.46","doi-asserted-by":"crossref","first-page":"919","DOI":"10.4049\/jimmunol.159.2.919","article-title":"CD44 regulates phagocytosis of apoptotic neutrophil granulocytes, but not apoptotic lymphocytes, by human macrophages","volume":"159","year":"1997","journal-title":"J Immunol"},{"issue":"4","key":"2023100513550562000_2023.03.30.535001v3.47","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1016\/j.ajpath.2019.12.006","article-title":"Intercellular Adhesion Molecule 1 Functions as an Efferocytosis Receptor in Inflammatory Macrophages","volume":"190","year":"2020","journal-title":"Am J Pathol"},{"key":"2023100513550562000_2023.03.30.535001v3.48","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20051166"},{"key":"2023100513550562000_2023.03.30.535001v3.49","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1004134"},{"key":"2023100513550562000_2023.03.30.535001v3.50","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.02357"},{"key":"2023100513550562000_2023.03.30.535001v3.51","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2019.06.017"},{"key":"2023100513550562000_2023.03.30.535001v3.52","doi-asserted-by":"publisher","DOI":"10.1038\/srep35067"},{"key":"2023100513550562000_2023.03.30.535001v3.53","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2014.08.008"},{"key":"2023100513550562000_2023.03.30.535001v3.54","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jit247"},{"issue":"7","key":"2023100513550562000_2023.03.30.535001v3.55","doi-asserted-by":"crossref","first-page":"1406","DOI":"10.21037\/jtd.2016.05.22","article-title":"The role of danger-associated molecular patterns (DAMPs) in trauma and infections","volume":"8","year":"2016","journal-title":"J Thorac Dis"},{"key":"2023100513550562000_2023.03.30.535001v3.56","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1601520"},{"key":"2023100513550562000_2023.03.30.535001v3.57","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1201302"},{"key":"2023100513550562000_2023.03.30.535001v3.58","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00170-10"},{"key":"2023100513550562000_2023.03.30.535001v3.59","doi-asserted-by":"publisher","DOI":"10.1038\/nature06329"},{"key":"2023100513550562000_2023.03.30.535001v3.60","doi-asserted-by":"crossref","unstructured":"Hsiao, C.-C. , et al., Macrophages Do Not Express the Phagocytic Receptor BAI1\/ADGRB1. Frontiers in Immunology, 2019. 10(962).","DOI":"10.3389\/fimmu.2019.00962"},{"key":"2023100513550562000_2023.03.30.535001v3.61","doi-asserted-by":"publisher","DOI":"10.1096\/fj.08-121228"},{"key":"2023100513550562000_2023.03.30.535001v3.62","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1302553110"},{"key":"2023100513550562000_2023.03.30.535001v3.63","doi-asserted-by":"crossref","unstructured":"Zhao, Y. , et al., Identification of Molecular Determinants in iRhoms1 and 2 That Contribute to the Substrate Selectivity of Stimulated ADAM17. Int J Mol Sci, 2022. 23(21).","DOI":"10.3390\/ijms232112796"},{"key":"2023100513550562000_2023.03.30.535001v3.64","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E17-04-0261"},{"key":"2023100513550562000_2023.03.30.535001v3.65","doi-asserted-by":"crossref","unstructured":"Zhu, H. , et al., The expression and clinical significance of different forms of Mer receptor tyrosine kinase in systemic lupus erythematosus. J Immunol Res, 2014. 2014: p. 431896.","DOI":"10.1155\/2014\/431896"},{"key":"2023100513550562000_2023.03.30.535001v3.66","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.2007.00094.x"},{"key":"2023100513550562000_2023.03.30.535001v3.67","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01597-06"},{"issue":"4","key":"2023100513550562000_2023.03.30.535001v3.68","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1021\/acscentsci.1c00200","article-title":"Bacterial Peptidoglycan Fragments Differentially Regulate Innate Immune Signaling","volume":"7","year":"2021","journal-title":"ACS Cent Sci"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2023.03.30.535001","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T04:29:47Z","timestamp":1768451387000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2023.03.30.535001"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,3]]},"references-count":68,"URL":"https:\/\/doi.org\/10.1101\/2023.03.30.535001","relation":{},"subject":[],"published":{"date-parts":[[2023,4,3]]},"subtype":"preprint"}}