{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T13:43:15Z","timestamp":1787578995319,"version":"build-2736575974"},"reference-count":49,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T00:00:00Z","timestamp":1670889600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>T cells expressing a simian immunodeficiency (SIV)-specific chimeric antigen receptor (CAR) and the follicular homing molecule, CXCR5, were infused into antiretroviral therapy (ART) suppressed, SIV-infected rhesus macaques to assess their ability to localize to the lymphoid follicle and control the virus upon ART interruption. While the cells showed evidence of functionality, they failed to persist in the animals beyond 28 days. Development of anti-CAR antibodies could be responsible for the lack of persistence. Potential antigenic sites on the anti-SIV CAR used in these studies included domains 1 and 2 of CD4, the carbohydrate recognition domain (CRD) of mannose-binding lectin (MBL), and an extracellular domain of the costimulatory molecule, CD28, along with short linker sequences. Using a flow cytometry based assay and target cells expressing the CAR\/CXCR5 construct, we examined the serum of the CD4-MBL CAR\/CXCR5-T cell treated animals to determine that the animals had developed an anti-CAR antibody response after infusion. Binding sites for the anti-CAR antibodies were identified by using alternative CARs transduced into target cells and by preincubation of the target cells with a CD4 blocking antibody. All of the treated animals developed antibodies in their serum that bound to CD4-MBL CAR\/CXCR5 T cells and the majority were capable of inducing an ADCC response. The CD4 antibody-blocking assay suggests that the dominant immunogenic components of this CAR are the CD4 domains with a possible additional site of the CD28 domain with its linker. This study shows that an anti-drug antibody (ADA) response can occur even when using self-proteins, likely due to novel epitopes created by abridged self-proteins and\/or the self-domain of the CAR connection to a small non-self linker. While in our study, there was no statistically significant correlation between the ADA response and the persistence of the CD4-MBL CAR\/CXCR5-T cells in rhesus macaques, these findings suggest that the development of an ADA response could impact the long-term persistence of self-based CAR immunotherapies.<\/jats:p>","DOI":"10.3389\/fimmu.2022.1032537","type":"journal-article","created":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T12:39:17Z","timestamp":1670935157000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Development of an anti-CAR antibody response in SIV-infected rhesus macaques treated with CD4-MBL CAR\/CXCR5 T cells"],"prefix":"10.3389","volume":"13","author":[{"given":"Brianna C.","family":"Davey","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary S.","family":"Pampusch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily K.","family":"Cartwright","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hadia M.","family":"Abdelaal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva G.","family":"Rakasz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aaron","family":"Rendahl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward A.","family":"Berger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pamela J.","family":"Skinner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,12,13]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1111\/imr.12243","article-title":"Going viral: chimeric antigen receptor T-cell therapy for hematological malignancies","volume":"263","author":"Gill","year":"2015","journal-title":"Immunol Rev"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1707447","article-title":"Axicabtagene ciloleucel CAR T-cell therapy in refractory Large b-cell lymphoma","volume":"377","author":"Neelapu","year":"2017","journal-title":"N Engl J Med"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1709866","article-title":"Tisagenlecleucel in children and young adults with b-cell lymphoblastic leukemia","volume":"378","author":"Maude","year":"2018","journal-title":"N Engl J Med"},{"key":"B4","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1056\/NEJMoa1804980","article-title":"Tisagenlecleucel in adult relapsed or refractory diffuse Large b-cell lymphoma","volume":"380","author":"Schuster","year":"2019","journal-title":"N Engl J Med"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.aav5685","article-title":"Multispecific anti-HIV duoCAR-T cells display broad in vitro antiviral activity and potent in vivo elimination of HIV-infected cells in a humanized mouse model","volume":"11","author":"Anthony-Gonda","year":"2019","journal-title":"Sci Transl Med"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00852-16","article-title":"Chimeric antigen receptor T cells guided by the single-chain fv of a broadly neutralizing antibody specifically and effectively eradicate virus reactivated from latency in CD4 T lymphocytes isolated from HIV-1-Infected individuals receiving suppressive combined antiretroviral therapy","volume":"90","author":"Liu","year":"2016","journal-title":"J Virol"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1172\/JCI144486","article-title":"CCR5-edited CD4+ T cells augment HIV-specific immunity to enable post-rebound control of HIV replication","volume":"131","author":"Tebas","year":"2021","journal-title":"J Clin Invest"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006613","article-title":"Supraphysiologic control over HIV-1 replication mediated by CD8 T cells expressing a re-engineered CD4-based chimeric antigen receptor","volume":"13","author":"Leibman","year":"2017","journal-title":"PloS Pathog"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcyt.2017.11.001","article-title":"Bispecific chimeric antigen receptors targeting the CD4 binding site and high-mannose glycans of gp120 optimized for anti-human immunodeficiency virus potency and breadth with minimal immunogenicity","volume":"20","author":"Ghanem","year":"2018","journal-title":"Cytotherapy"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.178.11.6975","article-title":"CTL fail to accumulate at sites of HIV-1 replication in lymphoid tissue","volume":"178","author":"Connick","year":"2007","journal-title":"J Immunol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1401161","article-title":"Compartmentalization of simian immunodeficiency virus replication within secondary lymphoid tissues of rhesus macaques is linked to disease stage and inversely related to localization of virus-specific CTL","volume":"193","author":"Connick","year":"2014","journal-title":"J Immunol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007311","article-title":"Low levels of SIV-specific CD8+ T cells in germinal centers characterizes acute SIV infection","volume":"15","author":"Li","year":"2019","journal-title":"PloS Pathog"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01332-16","article-title":"Simian immunodeficiency virus-producing cells in follicles are partially suppressed by CD8+ cells in vivo","volume":"90","author":"Li","year":"2016","journal-title":"J Virol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1186\/1742-4690-7-12","article-title":"In situ detection of gag-specific CD8+ cells in the GI tract of SIV infected rhesus macaques","volume":"7","author":"Tjernlund","year":"2010","journal-title":"Retrovirology"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1089\/aid.2005.21.363","article-title":"Lymphoid follicles are sites of heightened human immunodeficiency virus type 1 (HIV-1) replication and reduced antiretroviral effector mechanisms","volume":"21","author":"Folkvord","year":"2005","journal-title":"AIDS Res Hum Retroviruses"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1097\/00002030-199707000-00003","article-title":"Germinal centre CD4+ T cells are an important site of HIV replication in vivo","volume":"11","author":"Hufert","year":"1997","journal-title":"AIDS"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1084\/jem.187.6.949","article-title":"The unenlarged lymph nodes of HIV-1-infected, asymptomatic patients with high CD4 T cell counts are sites for virus replication and CD4 T cell proliferation. the impact of highly active antiretroviral therapy","volume":"187","author":"Tenner-Racz","year":"1998","journal-title":"J Exp Med"},{"key":"B18","article-title":"HTLV-III expression in infected lymph nodes and relevance to pathogenesis of lymphadenopathy","volume":"125","author":"Biberfeld","year":"1986","journal-title":"Am J Pathol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20121932","article-title":"Follicular helper T cells serve as the major CD4 T cell compartment for HIV-1 infection, replication, and production","volume":"210","author":"Perreau","year":"2013","journal-title":"J Exp Med"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2012-06-437608","article-title":"Differential infection patterns of CD4+ T cells and lymphoid tissue viral burden distinguish progressive and nonprogressive lentiviral infections","volume":"120","author":"Brenchley","year":"2012","journal-title":"Blood"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.00492","article-title":"Simian immunodeficiency virus (SIV)-specific chimeric antigen receptor-T cells engineered to target b cell follicles and suppress SIV replication","volume":"9","author":"Haran","year":"2018","journal-title":"Front Immunol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02507-16","article-title":"CXCR5-dependent entry of CD8 T cells into rhesus macaque b-cell follicles achieved through T-cell engineering","volume":"91","author":"Ayala","year":"2017","journal-title":"J Virol"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009831","article-title":"CAR\/CXCR5-T cell immunotherapy is safe and potentially efficacious in promoting sustained remission of SIV infection","volume":"18","author":"Pampusch","year":"2022","journal-title":"PloS Pathog"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006753","article-title":"Long-term persistence and function of hematopoietic stem cell-derived chimeric antigen receptor T cells in a nonhuman primate model of HIV\/AIDS","volume":"13","author":"Zhen","year":"2017","journal-title":"PloS Pathog"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-04390-6","article-title":"Decade-long leukaemia remissions with persistence of CD4+ CAR T cells","volume":"602","author":"Melenhorst","year":"2022","journal-title":"Nature"},{"key":"B26","doi-asserted-by":"publisher","first-page":"132ra53","DOI":"10.1126\/scitranslmed.3003761","article-title":"Decade-long safety and function of retroviral-modified chimeric antigen receptor T cells","volume":"4","author":"Scholler","year":"2012","journal-title":"Sci Transl Med"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.873701","article-title":"Evolving strategies to eliminate the CD4 T cells HIV viral reservoir via CAR T cell immunotherapy","volume":"13","author":"York","year":"2022","journal-title":"Front Immunol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1007\/s40259-019-00354-5","article-title":"Immunogenicity of chimeric antigen receptor T-cell therapeutics","volume":"33","author":"Gorovits","year":"2019","journal-title":"BioDrugs"},{"key":"B29","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1158\/2326-6066.cir-13-0006","article-title":"June CH. T cells expressing chimeric antigen receptors can cause anaphylaxis in humans","volume":"1","author":"Maus","year":"2013","journal-title":"Cancer Immunol Res"},{"key":"B30","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1182\/blood-2010-07-294520","article-title":"Immune responses to transgene and retroviral vector in patients treated with ex vivo\u2013engineered T cells","volume":"117","author":"Lamers","year":"2011","journal-title":"Blood"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2021.06.008","article-title":"Immune inactivation of anti-simian immunodeficiency virus chimeric antigen receptor T cells in rhesus macaques","volume":"22","author":"Haeseleer","year":"2021","journal-title":"Mol Ther Methods Clin Dev"},{"key":"B32","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.omtm.2019.09.007","article-title":"Rapid transduction and expansion of transduced T cells with maintenance of central memory populations","volume":"16","author":"Pampusch","year":"2020","journal-title":"Mol Ther - Methods Clin Dev"},{"key":"B33","first-page":"20130287748A1","article-title":"Use of chimeric antigen receptor-modified T cells to treat cancer","author":"June","year":"2013","journal-title":"US Patent"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2019.112692","article-title":"A cell-based immunogenicity assay to detect antibodies against chimeric antigen receptor expressed by tisagenlecleucel","volume":"476","author":"Potthoff","year":"2020","journal-title":"J Immunol Methods"},{"key":"B35","unstructured":"R: A language and environment for statistical computing2021"},{"key":"B36","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1007\/978-1-59745-450-6_1","article-title":"What is a b-cell epitope","volume":"524","author":"Van Regenmortel","year":"2009","journal-title":"Methods Mol Biol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1110\/ps.073347208","article-title":"The epitope space of the human proteome","volume":"17","author":"Berglund","year":"2008","journal-title":"Protein Sci"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1110\/ps.062405906","article-title":"Prediction of residues in discontinuous b-cell epitopes using protein 3D structures","volume":"15","author":"Andersen","year":"2006","journal-title":"Protein Sci"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00474-15","article-title":"Novel CD4-based bispecific chimeric antigen receptor designed for enhanced anti-HIV potency and absence of HIV entry receptor activity","volume":"89","author":"Liu","year":"2015","journal-title":"J Virol"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1003040107","article-title":"3D visualization of HIV transfer at the virological synapse between dendritic cells and T cells","volume":"107","author":"Felts","year":"2010","journal-title":"Proc Natl Acad Sci U.S.A."},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-020-1039-5","article-title":"Dual CD4-based CAR T cells with distinct costimulatory domains mitigate HIV pathogenesis in vivo","volume":"26","author":"Maldini","year":"2020","journal-title":"Nat Med"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1038\/sj.leu.2404737","article-title":"The dynamics of the b follicle: understanding the normal counterpart of b-cell-derived malignancies","volume":"21","author":"Sagaert","year":"2007","journal-title":"Leukemia"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.886546","article-title":"Immunogenicity of CAR-T cell therapeutics: Evidence, mechanism and mitigation","volume":"13","author":"Khan","year":"2022","journal-title":"Front Immunol"},{"key":"B44","unstructured":""},{"key":"B45","unstructured":""},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1016\/j.xphs.2018.12.014","article-title":"Immunogenicity of protein pharmaceuticals","volume":"108","author":"Dingman","year":"2019","journal-title":"J Pharm Sci"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1097\/CJI.0b013e31827806e6","article-title":"Sorting through subsets: which T-cell populations mediate highly effective adoptive immunotherapy","volume":"35","author":"Klebanoff","year":"2012","journal-title":"J Immunother"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1097\/COH.0000000000000485","article-title":"Chimeric antigen receptor T-cell approaches to HIV cure","volume":"13","author":"Kuhlmann","year":"2018","journal-title":"Curr Opin HIV AIDS"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2020006372","article-title":"Robust expansion of HIV CAR T cells following antigen boosting in ART-suppressed nonhuman primates","volume":"136","author":"Rust","year":"2020","journal-title":"Blood"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.1032537\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T12:39:23Z","timestamp":1670935163000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.1032537\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,13]]},"references-count":49,"alternative-id":["10.3389\/fimmu.2022.1032537"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2022.1032537","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,13]]},"article-number":"1032537"}}