{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T13:56:11Z","timestamp":1793109371412,"version":"4.5.0"},"reference-count":56,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,5,30]],"date-time":"2023-05-30T00:00:00Z","timestamp":1685404800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001711","name":"Schweizerischer Nationalfonds zur F\u00f6rderung der Wissenschaftlichen Forschung","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005688","name":"Gottfried und Julia Bangerter-Rhyner-Stiftung","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005688","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p>The larval stage of the cestode <jats:italic>Echinococcus multilocularis<\/jats:italic> is the causative agent of alveolar echinococcosis. To investigate the biology of these stages and to test novel compounds, metacestode cultures represent a suitable <jats:italic>in vitro<\/jats:italic> model system. These metacestodes are vesicles surrounded by an envelope formed by the vesicle tissue (VT), which is formed by the laminated and germinal layer, and filled with vesicle fluid (VF). We analyzed the proteome of VF and VT by liquid chromatography tandem mass spectrometry (LC-MS\/MS) and identified a total of 2,954 parasite proteins. The most abundant protein in VT was the expressed conserved protein encoded by EmuJ_000412500, followed by the antigen B subunit AgB8\/3a encoded by EmuJ_000381500 and Endophilin B1 (protein p29). In VF, the pattern was different and dominated by AgB subunits. The most abundant protein was the AgB8\/3a subunit followed by three other AgB subunits. In total, the AgB subunits detected in VF represented 62.1% of the parasite proteins. In culture media (CM), 63 <jats:italic>E. multilocularis<\/jats:italic> proteins were detected, of which AgB subunits made up 93.7% of the detected parasite proteins. All AgB subunits detected in VF (encoded by EmuJ_000381100\u2013700, corresponding to AgB8\/2, AgB8\/1, AgB8\/4, AgB8\/3a, AgB8\/3b, and AgB8\/3c) were also found in CM, except the subunit encoded by EmuJ_000381800 (AgB8\/5) that was very rare in VF and not detected in CM. The relative abundance of the AgB subunits in VF and CM followed the same pattern. In VT, only the subunits EmuJ_000381500 (AgB8\/3a) and EmuJ_000381200 (AgB8\/1) were detected among the 20 most abundant proteins. To see whether this pattern was specific to VF from <jats:italic>in vitro<\/jats:italic> cultured metacestodes, we analyzed the proteome of VF from metacestodes grown in a mouse model. Here, the AgB subunits encoded by EmuJ_000381100\u2013700 constituted the most abundant proteins, namely, 81.9% of total protein, with the same order of abundance as <jats:italic>in vitro<\/jats:italic>. Immunofluorescence on metacestodes showed that AgB is co-localized to calcareous corpuscles of <jats:italic>E. multilocularis<\/jats:italic>. Using targeted proteomics with HA-tagged EmuJ_000381200 (AgB8\/1) and EmuJ_000381100 (AgB8\/2), we could show that uptake of AgB subunits from CM into VF occurs within hours.<\/jats:p>","DOI":"10.3389\/fcimb.2023.1170763","type":"journal-article","created":{"date-parts":[[2023,5,31]],"date-time":"2023-05-31T13:07:25Z","timestamp":1685538445000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["Targeted and non-targeted proteomics to characterize the parasite proteins of Echinococcus multilocularis metacestodes"],"prefix":"10.3389","volume":"13","author":[{"given":"Joachim","family":"M\u00fcller","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mat\u00edas","family":"Preza","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc","family":"Kaethner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reto","family":"Rufener","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Braga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne-Christine","family":"Uldry","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manfred","family":"Heller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Britta","family":"Lundstr\u00f6m-Stadelmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,5,30]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"729","DOI":"10.1016\/0076-6879(91)94054-g","article-title":"Staining of actin with fluorochrome-conjugated phalloidin","volume":"194","author":"Adams","year":"1991","journal-title":"Methods Enzymol."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1186\/s13071-014-0610-7","article-title":"Alteration of immunoproteome profile of Echinococcus granulosus hydatid fluid with progression of cystic echinococcosis","volume":"8","author":"Ahn","year":"2015","journal-title":"Parasit. Vectors"},{"key":"B3","doi-asserted-by":"publisher","first-page":"438","DOI":"10.1186\/s13071-017-2359-2","article-title":"Fasciclin-calcareous corpuscle binary complex mediated protein-protein interactions in Taenia solium metacestode","volume":"10","author":"Ahn","year":"2017","journal-title":"Parasit. Vectors"},{"key":"B4","doi-asserted-by":"publisher","first-page":"806","DOI":"10.1021\/acs.jproteome.6b00799","article-title":"Biochemical characterization of Echinococcus multilocularis antigen B3 reveals insight into adaptation and maintenance of parasitic homeostasis at the host-parasite interface","volume":"16","author":"Ahn","year":"2017","journal-title":"J. Proteome Res."},{"key":"B5","doi-asserted-by":"publisher","first-page":"97009","DOI":"10.18632\/oncotarget.20761","article-title":"Comparison of Echinococcus multilocularis and Echinococcus granulosus hydatid fluid proteome provides molecular strategies for specialized host-parasite interactions","volume":"8","author":"Ahn","year":"2017","journal-title":"Oncotarget"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1560","DOI":"10.1016\/j.jprot.2011.02.021","article-title":"Proteomic characterisation of Echinococcus granulosus hydatid cyst fluid from sheep, cattle and humans","volume":"74","author":"Aziz","year":"2011","journal-title":"J. Proteomics"},{"key":"B7","doi-asserted-by":"publisher","first-page":"5564","DOI":"10.1128\/IAI.00653-09","article-title":"Molecular characterization of EmABP, an apolipoprotein a-I binding protein secreted by the Echinococcus multilocularis metacestode","volume":"77","author":"Bernthaler","year":"2009","journal-title":"Infect. Immun."},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES.2018.23.9.17-00161","article-title":"Prioritisation of food-borne parasites in europ","volume":"23","author":"Bouwknegt","year":"2018","journal-title":"Euro. Surveill."},{"key":"B9","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1007\/s00430-010-0154-1","article-title":"The role of evolutionarily conserved signalling systems in echinococcus multilocularis development and host-parasite interaction","volume":"199","author":"Brehm","year":"2010","journal-title":"Med. Microbiol. Immunol."},{"key":"B10","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1016\/bs.apar.2016.09.001","article-title":"Echinococcus-host interactions at cellular and molecular levels","volume":"95","author":"Brehm","year":"2017","journal-title":"Adv. Parasitol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1186\/s13071-018-2764-1","article-title":"Environmental risk factors and changing spatial patterns of human seropositivity for echinococcus spp. in xiji county, ningxia hui autonomous region, China","volume":"11","author":"Cadavid Restrepo","year":"2018","journal-title":"Parasit. Vectors"},{"key":"B12","doi-asserted-by":"publisher","first-page":"2327","DOI":"10.1042\/BCJ20190342","article-title":"Molecular mechanism of fast endophilin-mediated endocytosis","volume":"477","author":"Casamento","year":"2020","journal-title":"Biochem. J."},{"key":"B13","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1016\/j.pt.2019.05.005","article-title":"Echinococcus multilocularis","volume":"35","author":"Casulli","year":"2019","journal-title":"Trends Parasitol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"230","DOI":"10.3201\/eid2402.161126","article-title":"Echinococcus spp. tapeworms in north America","volume":"24","author":"Cerda","year":"2018","journal-title":"Emerg. Infect. Dis."},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot4947","article-title":"Labeling cytoskeletal f-actin with rhodamine phalloidin or fluorescein phalloidin for imaging","volume":"2010","author":"Chazotte","year":"2010","journal-title":"Cold Spring Harb. Protoc."},{"key":"B16","doi-asserted-by":"publisher","first-page":"315","DOI":"10.1016\/bs.apar.2016.11.001","article-title":"Global distribution of alveolar and cystic echinococcosis","volume":"95","author":"Deplazes","year":"2017","journal-title":"Adv. Parasitol."},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0005250","article-title":"Characterisation of antigen b protein species present in the hydatid cyst fluid of echinococcus canadensis G7 genotype","volume":"11","author":"Folle","year":"2017","journal-title":"PloS Negl. Trop. Dis."},{"key":"B18","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1016\/j.exppara.2008.03.002","article-title":"Echinococcus multilocularis: the parasite-host interplay","volume":"119","author":"Gottstein","year":"2008","journal-title":"Exp. Parasitol."},{"key":"B1000","doi-asserted-by":"publisher","first-page":"556","DOI":"10.1186\/1471-2164-13-556","article-title":"Proteome remodelling during development from blood to insect-form Trypanosoma brucei quantified by SILAC and mass spectrometry","volume":"13","author":"Gunasekera","year":"2012","journal-title":"BMC Genomics"},{"key":"B19","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1017\/S003118202100113X","article-title":"Echinococcus - the model cestode parasite","volume":"148","author":"Hemphill","year":"2021","journal-title":"Parasitology"},{"key":"B20","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1017\/S0022149X0928936X","article-title":"Alveolar and cystic echinococcosis: towards novel chemotherapeutical treatment options","volume":"83","author":"Hemphill","year":"2009","journal-title":"J. Helminthol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1051\/parasite\/2014073","article-title":"Treatment of echinococcosis: albendazole and mebendazole\u2013what else","volume":"21","author":"Hemphill","year":"2014","journal-title":"Parasite"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0004634","article-title":"Analysis on gene expression profile in oncospheres and early stage metacestodes from Echinococcus multilocularis","volume":"10","author":"Huang","year":"2016","journal-title":"PloS Negl. Trop. Dis."},{"key":"B23","doi-asserted-by":"publisher","first-page":"605","DOI":"10.1590\/0037-8682-0154-2013","article-title":"Analysis of the chemical components of hydatid fluid from Echinococcus granulosus","volume":"46","author":"Juyi","year":"2013","journal-title":"Rev. Soc. Bras. Med. Trop."},{"key":"B24","doi-asserted-by":"publisher","first-page":"259","DOI":"10.1016\/bs.apar.2016.09.006","article-title":"The echinococcoses: diagnosis, clinical management and burden of disease","volume":"96","author":"Kern","year":"2017","journal-title":"Adv. Parasitol."},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1186\/1742-9994-10-24","article-title":"Anatomy and development of the larval nervous system in Echinococcus multilocularis","volume":"10","author":"Koziol","year":"2013","journal-title":"Front. Zool."},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1186\/2041-9139-5-10","article-title":"The unique stem cell system of the immortal larva of the human parasite Echinococcus multilocularis","volume":"5","author":"Koziol","year":"2014","journal-title":"Evodevo"},{"key":"B27","doi-asserted-by":"publisher","first-page":"4016","DOI":"10.1002\/pmic.200700332","article-title":"A hydrophobic ligand-binding protein of the Taenia solium metacestode mediates uptake of the host lipid: implication for the maintenance of parasitic cellular homeostasis","volume":"7","author":"Lee","year":"2007","journal-title":"Proteomics"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1016\/j.fawpar.2019.e00040","article-title":"The importance of being parasiticidal. an update on drug development for the treatment of alveolar echinococcosis","volume":"15","author":"Lundstr\u00f6m-Stadelmann","year":"2019","journal-title":"Food Waterborne Parasitol."},{"key":"B1002","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/j.exppara.2005.12.007","article-title":"Echinococcus multilocularis: developmental stage-specific expression of Antigen B 8-kDa-subunits","volume":"113","author":"Mamuti","year":"2006","journal-title":"Exp. Parasitol."},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001551","article-title":"Echinococcus granulosus antigen b structure: subunit composition and oligomeric states","volume":"6","author":"Monteiro","year":"2012","journal-title":"PLoS Negl. Trop. Dis."},{"key":"B30","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1016\/j.jprot.2017.04.009","article-title":"Comparative proteomics of hydatid fluids from two echinococcus multilocularis isolates","volume":"162","author":"Monteiro","year":"2017","journal-title":"J. Proteomics"},{"key":"B31","doi-asserted-by":"publisher","first-page":"156","DOI":"10.1016\/j.exppara.2007.07.004","article-title":"Echinococcus granulosus antigen b gene family: further studies of strain polymorphism at the genomic and transcriptional levels","volume":"118","author":"Muzulin","year":"2008","journal-title":"Exp. Parasitol."},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001642","article-title":"Characterisation of the native lipid moiety of echinococcus granulosus antigen b","volume":"6","author":"Obal","year":"2012","journal-title":"PLoS Negl. Trop. Dis."},{"key":"B33","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1186\/s13071-016-1746-4","article-title":"The geographical distribution and prevalence of echinococcus multilocularis in animals in the European union and adjacent countries: a systematic review and meta-analysis","volume":"9","author":"Oksanen","year":"2016","journal-title":"Parasit. Vectors"},{"key":"B34","doi-asserted-by":"publisher","first-page":"130","DOI":"10.1111\/j.1365-3024.2011.01319.x","article-title":"Cestode genomics - progress and prospects for advancing basic and applied aspects of flatworm biology","volume":"34","author":"Olson","year":"2012","journal-title":"Parasite Immunol."},{"key":"B1001","doi-asserted-by":"publisher","first-page":"D543","DOI":"10.1093\/nar\/gkab1038","article-title":"The PRIDE database resources in 2022: a hub for mass spectrometry-based proteomics evidences","volume":"50","author":"Perez-Riverol","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"B35","doi-asserted-by":"publisher","first-page":"1665","DOI":"10.1002\/jcp.24266","article-title":"Alpha-2-Macroglobulin: a physiological guardian","volume":"228","author":"Rehman","year":"2013","journal-title":"J. Cell Physiol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1017\/s0022149x00007719","article-title":"Immunohistological localisation of two hydatid antigens (antigen 5 and antigen b) in the cyst wall, brood capsules and protoscoleces of Echinococcus granulosus (ovine and equine) and E. multilocularis using immunoperoxidase methods","volume":"51","author":"Rickard","year":"1977","journal-title":"J. Helminthol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"288","DOI":"10.1128\/IAI.69.1.288-296.2001","article-title":"Modulation of human immune response by Echinococcus granulosus antigen b and its possible role in evading host defenses","volume":"69","author":"Rigano","year":"2001","journal-title":"Infect. Immun."},{"key":"B38","doi-asserted-by":"publisher","first-page":"19438","DOI":"10.1038\/s41598-019-56073-y","article-title":"In vitro metabolomic footprint of the Echinococcus multilocularis metacestode","volume":"9","author":"Ritler","year":"2019","journal-title":"Sci. Rep."},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0009428","article-title":"Deep amplicon sequencing highlights low intra-host genetic variability of echinococcus multilocularis and high prevalence of the European-type haplotypes in coyotes and red foxes in Alberta, Canada","volume":"15","author":"Santa","year":"2021","journal-title":"PloS Negl. Trop. Dis."},{"key":"B40","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1016\/j.ijpara.2016.07.009","article-title":"Excretory\/secretory products in the Echinococcus granulosus metacestode: is the intermediate host complacent with infection caused by the larval form of the parasite","volume":"46","author":"Santos","year":"2016","journal-title":"Int. J. Parasitol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"B42","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1016\/bs.apar.2016.09.003","article-title":"Laboratory diagnosis of echinococcus spp. in human patients and infected animals","volume":"96","author":"Siles-Lucas","year":"2017","journal-title":"Adv. Parasitol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1186\/s13071-016-1350-7","article-title":"Echinococcus granulosus antigen b binds to monocytes and macrophages modulating cell response to inflammation","volume":"9","author":"Silva-Alvarez","year":"2016","journal-title":"Parasit. Vectors"},{"key":"B44","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1016\/j.plefa.2014.09.008","article-title":"Echinococcus granulosus antigen b: a hydrophobic ligand binding protein at the host-parasite interface","volume":"93","author":"Silva-Alvarez","year":"2015","journal-title":"Prostaglandins Leukot. Essent. Fatty Acids"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003552","article-title":"Lipid-free antigen b subunits from echinococcus granulosus: oligomerization, ligand binding, and membrane interaction properties","volume":"9","author":"Silva-Alvarez","year":"2015","journal-title":"PLoS Negl. Trop. Dis."},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1111\/pim.12575","article-title":"Antigen b from echinococcus granulosus is a novel ligand for c-reactive protein","volume":"40","author":"Silva-Alvarez","year":"2018","journal-title":"Parasite Immunol."},{"key":"B47","doi-asserted-by":"publisher","first-page":"1371","DOI":"10.1016\/j.ijpara.2008.06.003","article-title":"Molecular cross-talk in host-parasite relationships: the intriguing immunomodulatory role of echinococcus antigen b in cystic echinococcosis","volume":"38","author":"Siracusano","year":"2008","journal-title":"Int. J. Parasitol."},{"key":"B48","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1016\/j.exppara.2008.01.016","article-title":"Immunomodulatory mechanisms during Echinococcus granulosus infection","volume":"119","author":"Siracusano","year":"2008","journal-title":"Exp. Parasitol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"947","DOI":"10.1038\/cdd.2009.19","article-title":"Bif-1\/endophilin B1: a candidate for crescent driving force in autophagy","volume":"16","author":"Takahashi","year":"2009","journal-title":"Cell Death Differ"},{"key":"B50","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1016\/bs.apar.2016.07.001","article-title":"Biology and systematics of Echinococcus","volume":"95","author":"Thompson","year":"2017","journal-title":"Adv. Parasitol."},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1002\/prca.201700010","article-title":"Screening of antigenic vesicular fluid proteins of echinococcus multilocularis as potential viability biomarkers to monitor drug response in alveolar echinococcosis patients","volume":"11","author":"Valot","year":"2017","journal-title":"Proteomics Clin. Appl."},{"key":"B52","doi-asserted-by":"publisher","first-page":"517","DOI":"10.1016\/j.rvsc.2020.11.010","article-title":"Echinococcus granulosus sensu lato and Echinococcus multilocularis: a review","volume":"135","author":"Woolsey","year":"2021","journal-title":"Res. Vet. Sci."},{"key":"B53","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.vetpar.2017.01.012","article-title":"Regulatory effects of echinococcus multilocularis extracellular vesicles on RAW264.7 macrophages","volume":"235","author":"Zheng","year":"2017","journal-title":"Vet. Parasitol."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1170763\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,31]],"date-time":"2023-05-31T13:07:33Z","timestamp":1685538453000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1170763\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,5,30]]},"references-count":56,"alternative-id":["10.3389\/fcimb.2023.1170763"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2023.1170763","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,5,30]]},"article-number":"1170763"}}