{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T17:47:48Z","timestamp":1783964868530,"version":"3.55.0"},"reference-count":52,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2025,5,24]],"date-time":"2025-05-24T00:00:00Z","timestamp":1748044800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100021778","name":"Agencia Nacional de Promoci\u00f3n de la Investigaci\u00f3n, el Desarrollo Tecnol\u00f3gico y la Innovaci\u00f3n","doi-asserted-by":"publisher","award":["PICT 2020\u20103047"],"award-info":[{"award-number":["PICT 2020\u20103047"]}],"id":[{"id":"10.13039\/501100021778","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2025,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Shiga toxin\u2010producing\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    \u2010associated hemolytic uremic syndrome (STEC\u2010HUS) is a serious disease that causes renal failure predominantly in children. Despite its significant impact, there are currently no licensed vaccines or effective therapies available. The B subunits of Shiga toxins 1 and 2 (Stx1B and Stx2B) are suitable targets for developing neutralizing antibodies, but their pentameric assembly is unstable when isolated from the whole toxin. Taking advantage of the oligomeric symmetry shared between Stx1B and Stx2B with the lumazine synthase from\n                    <jats:italic>Brucella<\/jats:italic>\n                    spp. (BLS), we have previously engineered the chimeric toxoids BLS\u2010Stx1B and BLS\u2010Stx2B as immunogens to generate therapeutic equine polyclonal antibodies. The resulting product (INM004) has successfully passed Phases 1 and 2 clinical trials, and a Phase 3 has been launched in Argentina and seven European countries. In this work, we present the cryo\u2010electron microscopy structures of BLS\u2010Stx1B and BLS\u2010Stx2B, which confirm that these engineered immunogens effectively stabilize the StxB pentamers. Moreover, our results reveal that both chimeric constructs present high flexibility at their extremes, corresponding to motions of the StxBs with respect to the BLS core. Additionally, we present structural evidence of the interaction between the chimeras and polyclonal Fab (pFab) fragments derived from INM004, demonstrating that the elicited neutralizing antibodies block most of the interaction surface of the toxins with their cellular receptors. These findings further validate this promising antibody\u2010based therapy for mitigating STEC\u2010HUS and demonstrate that the BLS\u2010Stx1B and BLS\u2010Stx2B chimeras are potential candidates for developing a human vaccine.\n                  <\/jats:p>","DOI":"10.1002\/pro.70178","type":"journal-article","created":{"date-parts":[[2025,5,24]],"date-time":"2025-05-24T08:20:32Z","timestamp":1748074832000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Cryo\u2010\n                    <scp>EM<\/scp>\n                    structures of engineered Shiga toxin\u2010based immunogens capable of eliciting neutralizing antibodies with therapeutic potential against hemolytic uremic syndrome"],"prefix":"10.1002","volume":"34","author":[{"given":"Alejandro Ezequiel","family":"Crist\u00f3falo","sequence":"first","affiliation":[{"name":"Centro de Redise\u00f1o e Ingenier\u00eda de Prote\u00ednas (CRIP), Escuela de Bio y Nanotecnolog\u00edas (EByN), Universidad Nacional de San Mart\u00edn (UNSAM), Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET)  Gral. San Mart\u00edn Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arvind","family":"Sharma","sequence":"additional","affiliation":[{"name":"Genome Integrity and Structural Biology Laboratory National Institute of Environmental Health Sciences (NIEHS), National Institute of Health (NIH)  Research Triangle Park North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda Laura","family":"Cerutti","sequence":"additional","affiliation":[{"name":"Centro de Redise\u00f1o e Ingenier\u00eda de Prote\u00ednas (CRIP), Escuela de Bio y Nanotecnolog\u00edas (EByN), Universidad Nacional de San Mart\u00edn (UNSAM), Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET)  Gral. San Mart\u00edn Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kedar","family":"Sharma","sequence":"additional","affiliation":[{"name":"Genome Integrity and Structural Biology Laboratory National Institute of Environmental Health Sciences (NIEHS), National Institute of Health (NIH)  Research Triangle Park North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto","family":"Melero","sequence":"additional","affiliation":[{"name":"Centro Nacional de Biotecnolog\u00eda (CNB), Consejo Superior de Investigaciones Cient\u00edficas (CSIC)  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Romina","family":"Pardo","sequence":"additional","affiliation":[{"name":"Inmunova S.A  Gral. San Mart\u00edn Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando Alberto","family":"Goldbaum","sequence":"additional","affiliation":[{"name":"Centro de Redise\u00f1o e Ingenier\u00eda de Prote\u00ednas (CRIP), Escuela de Bio y Nanotecnolog\u00edas (EByN), Universidad Nacional de San Mart\u00edn (UNSAM), Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET)  Gral. San Mart\u00edn Buenos Aires Argentina"},{"name":"Inmunova S.A  Gral. San Mart\u00edn Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mario","family":"Borgnia","sequence":"additional","affiliation":[{"name":"Genome Integrity and Structural Biology Laboratory National Institute of Environmental Health Sciences (NIEHS), National Institute of Health (NIH)  Research Triangle Park North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanesa","family":"Zylberman","sequence":"additional","affiliation":[{"name":"Inmunova S.A  Gral. San Mart\u00edn Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5448-5483","authenticated-orcid":false,"given":"Lisandro Horacio","family":"Otero","sequence":"additional","affiliation":[{"name":"Centro de Redise\u00f1o e Ingenier\u00eda de Prote\u00ednas (CRIP), Escuela de Bio y Nanotecnolog\u00edas (EByN), Universidad Nacional de San Mart\u00edn (UNSAM), Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET)  Gral. San Mart\u00edn Buenos Aires Argentina"},{"name":"Departamento de Biolog\u00eda Molecular, Facultad de Ciencias Exactas, F\u00edsico\u2010Qu\u00edmicas y Naturales Instituto de Biotecnolog\u00eda Ambiental y Salud (INBIAS), Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET), Universidad Nacional de R\u00edo Cuarto (UNRC)  R\u00edo Cuarto C\u00f3rdoba Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,5,24]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909052925"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-25087-4"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.5546\/AAP.2021.238"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00467-023-06104-8"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abn2911"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0575-8"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2018.07.009"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pcl.2022.07.006"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.2000.3640"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.medmal.2017.09.012"},{"key":"e_1_2_9_12_1","first-page":"1","volume-title":"Methods in Enzymology","author":"Ceballos\u2010Alcantarilla E","year":"2021"},{"key":"e_1_2_9_13_1","unstructured":"ClinicalTrails.gov.A phase III study to evaluate the efficacy of INM004 (Shiga antitoxin) in pediatric patients with Shiga toxin\u2010producing Escherichia coli\u2010associated hemolytic uremic syndrome. 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