Uptake of bacteria by phagocytes is a crucial step in innate immune defence. Members of the disintegrin and metalloproteinase (ADAM) family critically control the immune response by limited proteolysis of surface expressed mediator molecules. Here, we investigated the significance of ADAM17 and its regulatory adapter molecule iRhom2 for bacterial uptake by phagocytes. Inhibition of metalloproteinase activity led to increased phagocytosis of pHrodo labelled Gram-negative and -positive bacteria (E. coli and S. aureus, respectively) by human and murine monocytic cell lines or primary phagocytes. Bone marrow-derived macrophages showed enhanced uptake of heat-inactivated and living E. coli when they lacked either ADAM17 or iRhom2 but not upon ADAM10-deficiency. In monocytic THP-1 cells, corresponding short hairpin RNA (shRNA)-mediated knockdown confirmed that ADAM17, but not ADAM10, promoted phagocytosis of E. coli. The augmented bacterial uptake occurred in a cell autonomous manner and was accompanied by increased release of the chemokine CXCL8, less TNFα release and only minimal changes in the surface expression of the receptors TNFR1, TLR6 and CD36. Inhibition experiments indicated that the enhanced bacterial phagocytosis after ADAM17 knockdown was partially dependent on TNFα-activity but not on CXCL8. This novel role of ADAM17 in bacterial uptake needs to be considered in the development of ADAM17 inhibitors as therapeutics.

Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences 25250 Vestec Czech Republic

Department of Neonatology University Children's Hospital 52074 Aachen Germany

Institute of Experimental and Clinical Pharmacology and Toxicology PZMS ZHMB Saarland University 66424 Homburg Germany

Institute of Molecular Pharmacology Medical Faculty RWTH Aachen University 52074 Aachen Germany

Zobrazit více v PubMed

Dreymueller D., Uhlig S., Ludwig A. Adam-family metalloproteinases in lung inflammation: Potential therapeutic targets. Am. J. Physiol. Lung Cell. Mol. Physiol. 2015;308:325–343. doi: 10.1152/ajplung.00294.2014. PubMed DOI

Pruessmeyer J., Ludwig A. The good, the bad and the ugly substrates for ADAM10 and ADAM17 in brain pathology, inflammation and cancer. Semin. Cell Dev. Biol. 2009;20:164–174. doi: 10.1016/j.semcdb.2008.09.005. PubMed DOI

Düsterhöft S., Babendreyer A., Giese A.A., Flasshove C., Ludwig A. Status update on iRhom and ADAM17: It’s still complicated. Biochim. Biophys. Acta-Mol. Cell Res. 2019;1866:1567–1583. doi: 10.1016/j.bbamcr.2019.06.017. PubMed DOI

Chalaris A., Adam N., Sina C., Rosenstiel P., Lehmann-Koch J., Schirmacher P., Hartmann D., Cichy J., Gavrilova O., Schreiber S., et al. Critical role of the disintegrin metalloprotease ADAM17 for intestinal inflammation and regeneration in mice. J. Exp. Med. 2010;207:1617–1624. doi: 10.1084/jem.20092366. PubMed DOI PMC

Peschon J.J., Slack J.L., Reddy P., Stocking K.L., Sunnarborg S.W., Lee D.C., Russell W.E., Castner B.J., Johnson R.S., Fitzner J.N., et al. An essential role for ectodomain shedding in mammalian development. Science (80-) 1998;282:1281–1284. doi: 10.1126/science.282.5392.1281. PubMed DOI

Adrain C., Zettl M., Christova Y., Taylor N., Freeman M. Tumor necrosis factor signaling requires iRhom2 to promote trafficking and activation of TACE. Science (80-) 2012;335:225–228. doi: 10.1126/science.1214400. PubMed DOI PMC

McIlwain D.R., Lang P.A., Maretzky T., Hamada K., Ohishi K., Maney S.K., Berger T., Murthy A., Duncan G., Xu H.C., et al. iRhom2 regulation of TACE controls TNF-mediated protection against Listeria and responses to LPS. Science (80-) 2012;335:229–232. doi: 10.1126/science.1214448. PubMed DOI PMC

Li X., Maretzky T., Weskamp G., Monette S., Qing X., Issuree P.D.A., Crawford H.C., McIlwain D.R., Mak T.W., Salmon J.E., et al. iRhoms 1 and 2 are essential upstream regulators of ADAM17-dependent EGFR signaling. Proc. Natl. Acad. Sci. USA. 2015;112:6080–6085. doi: 10.1073/pnas.1505649112. PubMed DOI PMC

Christova Y., Adrain C., Bambrough P., Ibrahim A., Freeman M. Mammalian iRhoms have distinct physiological functions including an essential role in TACE regulation. EMBO Rep. 2013;14:884–890. doi: 10.1038/embor.2013.128. PubMed DOI PMC

Issuree P.D.A., Maretzky T., McIlwain D.R., Monette S., Qing X., Lang P.A., Swendeman S.L., Park-Min K.H., Binder N., Kalliolias G.D., et al. iRHOM2 is a critical pathogenic mediator of infammatory arthritis. J. Clin. Investig. 2013;123:928–932. doi: 10.1172/JCI66168. PubMed DOI PMC

Blaydon D.C., Etheridge S.L., Risk J.M., Hennies H.C., Gay L.J., Carroll R., Plagnol V., McRonald F.E., Stevens H.P., Spurr N.K., et al. RHBDF2 mutations are associated with tylosis, a familial esophageal cancer syndrome. Am. J. Hum. Genet. 2012;90:340–346. doi: 10.1016/j.ajhg.2011.12.008. PubMed DOI PMC

Tsukerman P., Eisenstein E.M., Chavkin M., Schmiedel D., Wong E., Werner M., Yaacov B., Averbuch D., Molho-Pessach V., Stepensky P., et al. Cytokine secretion and NK cell activity in human ADAM17 deficiency. Oncotarget. 2015;6:44151. doi: 10.18632/oncotarget.6629. PubMed DOI PMC

Blaydon D.C., Biancheri P., Di W.L., Plagnol V., Cabral R.M., Brooke M.A., Van Heel D.A., Ruschendorf F., Toynbee M., Walne A., et al. Inflammatory skin and bowel disease linked to ADAM17 deletion. N. Engl. J. Med. 2011;365:1502–1508. doi: 10.1056/NEJMoa1100721. PubMed DOI

Horiuchi K., Kimura T., Miyamoto T., Takaishi H., Okada Y., Toyama Y., Blobel C.P. Cutting Edge: TNF-α-Converting Enzyme (TACE/ADAM17) Inactivation in Mouse Myeloid Cells Prevents Lethality from Endotoxin Shock. J. Immunol. 2007;179:2686–2689. doi: 10.4049/jimmunol.179.5.2686. PubMed DOI

Dreymueller D., Martin C., Kogel T., Pruessmeyer J., Hess F.M., Horiuchi K., Uhlig S., Ludwig A. Lung endothelial ADAM17 regulates the acute inflammatory response to lipopolysaccharide. EMBO Mol. Med. 2012;4:412–423. doi: 10.1002/emmm.201200217. PubMed DOI PMC

Arndt P.G., Strahan B., Wang Y., Long C., Horiuchi K., Walcheck B. Leukocyte adam17 regulates acute pulmonary inflammation. PLoS ONE. 2011;6:e19938. doi: 10.1371/journal.pone.0019938. PubMed DOI PMC

Schulz B., Pruessmeyer J., Maretzky T., Ludwig A., Blobel C.P., Saftig P., Reiss K. ADAMIO regulates endothelial permeability and T-cell transmigration by proteolysis of vascular endothelial cadherin. Circ. Res. 2008;102:1192–1201. doi: 10.1161/CIRCRESAHA.107.169805. PubMed DOI PMC

Hundhausen C., Schulte A., Schulz B., Andrzejewski M.G., Schwarz N., von Hundelshausen P., Winter U., Paliga K., Reiss K., Saftig P., et al. Regulated Shedding of Transmembrane Chemokines by the Disintegrin and Metalloproteinase 10 Facilitates Detachment of Adherent Leukocytes. J. Immunol. 2007;178:8064–8072. doi: 10.4049/jimmunol.178.12.8064. PubMed DOI

Dreymueller D., Martin C., Schumacher J., Groth E., Boehm J.K., Reiss L.K., Uhlig S., Ludwig A. Smooth Muscle Cells Relay Acute Pulmonary Inflammation via Distinct ADAM17/ErbB Axes. J. Immunol. 2014;192:722–731. doi: 10.4049/jimmunol.1302496. PubMed DOI

Bell J.H., Herrera A.H., Li Y., Walcheck B. Role of ADAM17 in the ectodomain shedding of TNF- and its receptors by neutrophils and macrophages. J. Leukoc. Biol. 2007;82:173–176. doi: 10.1189/jlb.0307193. PubMed DOI

Li Y., Brazzell J., Herrera A., Walcheck B. ADAM17 deficiency by mature neutrophils has differential effects on L-selectin shedding. Blood. 2006;108:2275–2279. doi: 10.1182/blood-2006-02-005827. PubMed DOI PMC

Etzerodt A., Maniecki M.B., Møller K., Møller H.J., Moestrup S.K. Tumor necrosis factor α-converting enzyme (TACE/ADAM17) mediates ectodomain shedding of the scavenger receptor CD163. J. Leukoc. Biol. 2010;88:1201–1205. doi: 10.1189/jlb.0410235. PubMed DOI

Kneidl J., Löffler B., Erat M.C., Kalinka J., Peters G., Roth J., Barczyk K. Soluble CD163 promotes recognition, phagocytosis and killing of Staphylococcus aureus via binding of specific fibronectin peptides. Cell. Microbiol. 2012;14:914–936. doi: 10.1111/j.1462-5822.2012.01766.x. PubMed DOI

Driscoll W.S., Vaisar T., Tang J., Wilson C.L., Raines E.W. Macrophage ADAM17 deficiency augments CD36-dependent apoptotic cell uptake and the linked anti-inflammatory phenotype. Circ. Res. 2013;113:52–61. doi: 10.1161/CIRCRESAHA.112.300683. PubMed DOI PMC

Gough P.J., Garton K.J., Wille P.T., Rychlewski M., Dempsey P.J., Raines E.W. A Disintegrin and Metalloproteinase 10-Mediated Cleavage and Shedding Regulates the Cell Surface Expression of CXC Chemokine Ligand 16. J. Immunol. 2004;172:3678–3685. doi: 10.4049/jimmunol.172.6.3678. PubMed DOI

Langjahr P., Díaz-Jiménez D., De La Fuente M., Rubio E., Golenbock D., Bronfman F.C., Quera R., Lez M.J.G., Hermoso M.A., Benjamim C.F. Metalloproteinase-dependent TLR2 ectodomain shedding is involved in soluble toll-like receptor 2 (sTLR2) production. PLoS ONE. 2014;9:e104624. doi: 10.1371/journal.pone.0104624. PubMed DOI PMC

Richmond J.M., Duffy E.R., Lee J., Kaboli K., Remick D.G., Kornfeld H., Cruikshank W.W. Mannose-capped lipoarabinomannan from Mycobacterium tuberculosis induces soluble tumor necrosis factor receptor production through tumor necrosis factor alpha-converting enzyme activation. Infect. Immun. 2012;80:3858–3868. doi: 10.1128/IAI.00060-12. PubMed DOI PMC

Long C., Wang Y., Herrera A.H., Horiuchi K., Walcheck B. In vivo role of leukocyte ADAM17 in the inflammatory and host responses during E. coli -mediated peritonitis. J. Leukoc. Biol. 2010;87:1097–1101. doi: 10.1189/jlb.1109763. PubMed DOI PMC

Long C., Hosseinkhani M.R., Wang Y., Sriramarao P., Walcheck B. ADAM17 activation in circulating neutrophils following bacterial challenge impairs their recruitment. J. Leukoc. Biol. 2012;92:667–672. doi: 10.1189/jlb.0312112. PubMed DOI PMC

Sommer D., Corstjens I., Sanchez S., Dooley D., Lemmens S., Van Broeckhoven J., Bogie J., Vanmierlo T., Vidal P.M., Rose-John S., et al. ADAM17-deficiency on microglia but not on macrophages promotes phagocytosis and functional recovery after spinal cord injury. Brain. Behav. Immun. 2019;80:129–145. doi: 10.1016/j.bbi.2019.02.032. PubMed DOI

Black R.A., Rauch C.T., Kozlosky C.J., Peschon J.J., Slack J.L., Wolfson M.F., Castner B.J., Stocking K.L., Reddy P., Srinivasan S., et al. A metalloproteinase disintegrin that releases tumour-necrosis factor-∅ from cells. Nature. 1997;385:729–733. doi: 10.1038/385729a0. PubMed DOI

Schwarz M., Taubitz A., Eltrich N., Mulay S.R., Allam R., Vielhauer V. Analysis of TNF-mediated recruitment and activation of glomerular dendritic cells in mouse kidneys by compartment-specific flow cytometry. Kidney Int. 2013;84:116–129. doi: 10.1038/ki.2013.46. PubMed DOI

Matthews V., Schuster B., Schütze S., Bussmeyer I., Ludwig A., Hundhausen C., Sadowski T., Saftig P., Hartmann D., Kallen K.J., et al. Cellular cholesterol depletion triggers shedding of the human interleukin-6 receptor by ADAM10 and ADAM17 (TACE) J. Biol. Chem. 2003;278:38829–38839. doi: 10.1074/jbc.M210584200. PubMed DOI

Chalaris A., Gewiese J., Paliga K., Fleig L., Schneede A., Krieger K., Rose-John S., Scheller J. ADAM17-mediated shedding of the IL6R induces cleavage of the membrane stub by γ-secretase. Biochim. Biophys. Acta-Mol. Cell Res. 2010;1803:234–245. doi: 10.1016/j.bbamcr.2009.12.001. PubMed DOI

Friedland J.S., Constantin D., Shaw T.C., Stylianou E. Regulation of interleukin-8 gene expression after phagocytosis of zymosan by human monocytic cells. J. Leukoc. Biol. 2001;70:447–454. doi: 10.1189/jlb.70.3.447. PubMed DOI

Friedland J.S., Remick D.G., Shattock R., Griffin G.E. Secretion of interleukin-8 following phagocytosis of Mycobacterium tuberculosis by human monocyte cell lines. Eur. J. Immunol. 1992;22:1373–1378. doi: 10.1002/eji.1830220607. PubMed DOI

Kang H.J., Ha J.M., Kim H.S., Lee H., Kurokawa K., Lee B.L. The role of phagocytosis in IL-8 production by human monocytes in response to lipoproteins on Staphylococcus aureus. Biochem. Biophys. Res. Commun. 2011;406:449–453. doi: 10.1016/j.bbrc.2011.02.069. PubMed DOI

Zhang J., Li H., Wang J., Dong Z., Mian S., Yu F.S.X. Role of EGFR transactivation in preventing apoptosis in Pseudomonas aeruginosa-infected human corneal epithelial cells. Investig. Ophthalmol. Vis. Sci. 2004;45:2569–2576. doi: 10.1167/iovs.03-1323. PubMed DOI PMC

Yang W.S., Kim J.J., Lee M.J., Lee E.K., Park S.K. ADAM17-Mediated Ectodomain Shedding of Toll-Like Receptor 4 as a Negative Feedback Regulation in Lipopolysaccharide-Activated Aortic Endothelial Cells. Cell. Physiol. Biochem. 2018;45:1851–1862. doi: 10.1159/000487876. PubMed DOI

Xu P., Liu J., Sakaki-Yumoto M., Derynck R. TACE activation by MAPK-mediated regulation of cell surface dimerization and TIMP3 association. Sci. Signal. 2012;5:ra34. doi: 10.1126/scisignal.2002689. PubMed DOI PMC

Yoda M., Kimura T., Tohmonda T., Morioka H., Matsumoto M., Okada Y., Toyama Y., Horiuchi K. Systemic Overexpression of TNFα-converting Enzyme Does Not Lead to Enhanced Shedding Activity In Vivo. PLoS ONE. 2013;8:e54412. doi: 10.1371/journal.pone.0054412. PubMed DOI PMC

Düsterhöft S., Michalek M., Kordowski F., Oldefest M., Sommer A., Röseler J., Reiss K., Grötzinger J., Lorenzen I. Extracellular Juxtamembrane Segment of ADAM17 Interacts with Membranes and Is Essential for Its Shedding Activity. Biochemistry. 2015;54:5791–5801. doi: 10.1021/acs.biochem.5b00497. PubMed DOI

Pruessmeyer J., Hess F.M., Alert H., Groth E., Pasqualon T., Schwarz N., Nyamoya S., Kollert J., Van Der Vorst E., Donners M., et al. Leukocytes require ADAM10 but not ADAM17 for their migration and inflammatory recruitment into the alveolar space. Blood. 2014;123:4077–4088. doi: 10.1182/blood-2013-09-511543. PubMed DOI

Koenen R.R., Pruessmeyer J., Soehnlein O., Fraemohs L., Zernecke A., Schwarz N., Reiss K., Sarabi A., Lindbom L., Hackeng T.M., et al. Regulated release and functional modulation of junctional adhesion molecule A by disintegrin metalloproteinases. Blood. 2009;113:4799–4809. doi: 10.1182/blood-2008-04-152330. PubMed DOI

Pruessmeyer J., Martin C., Hess F.M., Schwarz N., Schmidt S., Kogel T., Hoettecke N., Schmidt B., Sechi A., Uhlig S., et al. A Disintegrin and metalloproteinase 17 (ADAM17) mediates inflammation-induced shedding of syndecan-1 and -4 by lung epithelial cells. J. Biol. Chem. 2010;285:555–564. doi: 10.1074/jbc.M109.059394. PubMed DOI PMC

Babendreyer A., Molls L., Simons I.M., Dreymueller D., Biller K., Jahr H., Denecke B., Boon R.A., Bette S., Schnakenberg U., et al. The metalloproteinase ADAM15 is upregulated by shear stress and promotes survival of endothelial cells. J. Mol. Cell. Cardiol. 2019;134:51–61. doi: 10.1016/j.yjmcc.2019.06.017. PubMed DOI

A structural model of the iRhom-ADAM17 sheddase complex reveals functional insights into its trafficking and activity

The iRhom homology domain is indispensable for ADAM17-mediated TNFα and EGF receptor ligand release