{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,25]],"date-time":"2026-10-25T12:39:26Z","timestamp":1792931966260,"version":"4.5.0"},"reference-count":18,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2024,5,2]],"date-time":"2024-05-02T00:00:00Z","timestamp":1714608000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["R35 CA210084 (JFD), NIH\/NCI: P50 CA171963 NCI (JFD) and NIH\/NCI R50 CA211466-06 (MPR)."],"award-info":[{"award-number":["R35 CA210084 (JFD), NIH\/NCI: P50 CA171963 NCI (JFD) and NIH\/NCI R50 CA211466-06 (MPR)."]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Off-the-shelf immunotherapeutics that suppress tumor growth and provide durable protection against relapse could enhance cancer treatment. We report preclinical studies on a CD33 x CD3 bivalent bispecific diabody, AMV564, that not only suppresses tumor growth, but also facilitates memory responses in a mouse model of acute myelogenous leukemia (AML). Mechanistically, a single 5-day treatment with AMV564 seems to reduce tumor burden by redirection of T cells, providing a time window for allogeneic or other T cells that innately recognize tumor antigens to become activated and proliferate. When the concentration of bispecific becomes negligible, the effector: target ratio has also shifted, and these activated T cells mediate long-term tumor control. To test the efficacy of AMV564 <jats:italic>in vivo<\/jats:italic>, we generated a CD33<jats:sup>+<\/jats:sup> MOLM13<jats:sup>CG<\/jats:sup> bioluminescent human cell line and optimized conditions needed to control these cells for 62 days <jats:italic>in vivo<\/jats:italic> in NSG mice. Of note, not only did MOLM13<jats:sup>CG<\/jats:sup> become undetectable by bioluminescence imaging in response to infusion of human T cells plus AMV564, but also NSG mice that had cleared the tumor also resisted rechallenge with MOLM13<jats:sup>CG<\/jats:sup> in spite of no additional AMV564 treatment. In these mice, we identified effector and effector memory human CD4<jats:sup>+<\/jats:sup> and CD8<jats:sup>+<\/jats:sup> T cells in the peripheral blood immediately prior to rechallenge that expanded significantly during the subsequent 18 days. In addition to the anti-tumor effects of AMV564 on the clearance of MOLM13<jats:sup>CG<\/jats:sup> cells <jats:italic>in vivo<\/jats:italic>, similar effects were seen when primary CD33<jats:sup>+<\/jats:sup> human AML cells were engrafted in NSG mice even when the human T cells made up only 2% of the peripheral blood cells and AML cells made up 98%. These studies suggest that AMV564 is a novel and effective bispecific diabody for the targeting of CD33<jats:sup>+<\/jats:sup> AML that may provide long-term survival advantages in the clinic.<\/jats:p>","DOI":"10.1371\/journal.pone.0300174","type":"journal-article","created":{"date-parts":[[2024,5,2]],"date-time":"2024-05-02T17:26:49Z","timestamp":1714670809000},"page":"e0300174","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":7,"title":["Control of acute myeloid leukemia and generation of immune memory in vivo using AMV564, a bivalent bispecific CD33 x CD3 T cell engager"],"prefix":"10.1371","volume":"19","author":[{"given":"Linda G.","family":"Eissenberg","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie K.","family":"Ritchey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael P.","family":"Rettig","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0765-4990","authenticated-orcid":true,"given":"Dilan A.","family":"Patel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kiran","family":"Vij","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Smith","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tae H.","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John F.","family":"DiPersio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,5,2]]},"reference":[{"issue":"6","key":"pone.0300174.ref001","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1007\/s11899-018-0472-8","article-title":"Bispecific Antibodies for the Treatment of Acute Myeloid Leukemia","volume":"13","author":"DG Guy","year":"2018","journal-title":"Curr Hematol Malig Rep"},{"issue":"23","key":"pone.0300174.ref002","doi-asserted-by":"crossref","first-page":"5829","DOI":"10.1158\/1078-0432.CCR-16-0350","article-title":"Characterization of CD33\/CD3 Tetravalent Bispecific Tandem Diabodies (TandAbs) for the Treatment of Acute Myeloid Leukemia","volume":"22","author":"U Reusch","year":"2016","journal-title":"Clin Cancer Res"},{"issue":"3","key":"pone.0300174.ref003","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1080\/19420862.2015.1029216","article-title":"A tetravalent bispecific TandAb (CD19\/CD3), AFM11, efficiently recruits T cells for the potent lysis of CD19(+) tumor cells","volume":"7","author":"U Reusch","year":"2015","journal-title":"MAbs"},{"issue":"supplement 1","key":"pone.0300174.ref004","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1182\/blood-2018-99-111529","article-title":"Phase 1 First-in-Human Trial of AMV64, a bivalent bisepcific (2x2) CD33\/CD3 T-Cell Engager, in patients with relapsed\/refractory Acute Myeloid Leukemia (AML)","volume":"132","author":"P Westervelt","year":"2018","journal-title":"Blood"},{"key":"pone.0300174.ref005","doi-asserted-by":"crossref","unstructured":"Westervelt P, Roboz G, Cortes J, Altman J, Oehler V, Long M, et al. Safety and clinical activity of AMV564, a CD33\/CD3 T-cell engager, in patients with relapsed\/refractory acute myeloiid leukemia (AML): updated results from the phase 1 first-in-human trial. 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Optimized twenty-one color panel design using the ZE5 cell analyzer for quantification of T cell subsets in stem cell transplant patients. 2017."},{"issue":"29","key":"pone.0300174.ref010","doi-asserted-by":"crossref","first-page":"3544","DOI":"10.1200\/JCO.2016.67.5892","article-title":"Extramedullary Disease in Adult Acute Myeloid Leukemia Is Common but Lacks Independent Significance: Analysis of Patients in ECOG-ACRIN Cancer Research Group Trials, 1980\u20132008","volume":"34","author":"C Ganzel","year":"2016","journal-title":"J Clin Oncol"},{"key":"pone.0300174.ref011","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1182\/blood-2020-133891","article-title":"Flotetuzumab and Other Cellular Immunotherapies Upregulate MHC Class II Expression on Acute Myeloid Leukemia Cells in Vitro and In Vivo","volume":"136","author":"J Rimando","year":"2020","journal-title":"Blood"},{"issue":"10","key":"pone.0300174.ref012","doi-asserted-by":"crossref","first-page":"1079","DOI":"10.1038\/nbt.3349","article-title":"Therapeutic bispecific T-cell engager antibody targeting the intracellular oncoprotein WT1","volume":"33","author":"T Dao","year":"2015","journal-title":"Nature biotechnology"},{"issue":"6","key":"pone.0300174.ref013","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1158\/1535-7163.MCT-13-0956","article-title":"Preclinical characterization of AMG 330, a CD3\/CD33-bispecific T-cell-engaging antibody with potential for treatment of acute myelogenous leukemia","volume":"13","author":"M Friedrich","year":"2014","journal-title":"Mol Cancer Ther"},{"issue":"23","key":"pone.0300174.ref014","doi-asserted-by":"crossref","first-page":"2484","DOI":"10.1182\/blood-2018-05-849802","article-title":"Bifunctional PD-1 x alphaCD3 x alphaCD33 fusion protein reverses adaptive immune escape in acute myeloid leukemia","volume":"132","author":"M Herrmann","year":"2018","journal-title":"Blood"},{"issue":"11","key":"pone.0300174.ref015","doi-asserted-by":"crossref","first-page":"1250","DOI":"10.1182\/bloodadvances.2017014373","article-title":"A potent tetravalent T-cell-engaging bispecific antibody against CD33 in acute myeloid leukemia","volume":"2","author":"SS Hoseini","year":"2018","journal-title":"Blood Adv"},{"issue":"5","key":"pone.0300174.ref016","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1038\/leu.2012.341","article-title":"T lymphocytes can be effectively recruited for ex vivo and in vivo lysis of AML blasts by a novel CD33\/CD3-bispecific BiTE antibody construct","volume":"27","author":"M Aigner","year":"2013","journal-title":"Leukemia"},{"issue":"1","key":"pone.0300174.ref017","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1002\/(SICI)1097-0215(19990105)80:1<138::AID-IJC25>3.0.CO;2-J","article-title":"Tumor-growth inhibition with bispecific antibody fragments in a syngeneic mouse melanoma model: the role of targeted T-cell co-stimulation via CD28","volume":"80","author":"L Grosse-Hovest","year":"1999","journal-title":"Int J Cancer"},{"issue":"6","key":"pone.0300174.ref018","doi-asserted-by":"crossref","first-page":"2066","DOI":"10.1158\/0008-5472.CAN-10-3200","article-title":"Cancer immunotherapy using a bispecific NK receptor fusion protein that engages both T cells and tumor cells","volume":"71","author":"T Zhang","year":"2011","journal-title":"Cancer research"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0300174","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,2]],"date-time":"2024-05-02T17:27:03Z","timestamp":1714670823000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0300174"}},"subtitle":[],"editor":[{"given":"Anilkumar","family":"Gopalakrishnapillai","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,5,2]]},"references-count":18,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5,2]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0300174","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,2]]}}}