{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T20:25:10Z","timestamp":1791923110301,"version":"4.3.4"},"reference-count":31,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2023,9,15]],"date-time":"2023-09-15T00:00:00Z","timestamp":1694736000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["European J of Haematology"],"published-print":{"date-parts":[[2024,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Objectives<\/jats:title><jats:p>The treatment of refractory and recurrent acute myeloid leukaemia (AML) is still a challenge with poor response rates and short survival times. In an attempt to solve this problem, we constructed a tandem bispecific chimeric antigen receptor (CAR) targeting CD123 and C\u2010type lectin\u2010like molecule 1 (CLL\u20101), two different AML antigens, and verified its cytotoxic effects in vitro.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We established and cultured K562 cell lines expressing both CD123 and CLL1 antigens. Single\u2010target CAR\u2010T cells specific to CD123 and CLL1 were engineered, alongside tandem CD123\/CLL1 bispecific CAR\u2010T cells. Flow cytometry was used to determine cell phenotypes, transfection efficiencies, cytokine release, and CAR\u2010T\u2010cell proliferation, and an lactate dehydrogenase assay was used to detect the cytotoxicity of CD123\/CLL\u20101 bispecific tandem CAR\u2010T cells in vitro.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Two types of tandem CAR\u2010T cells exhibited significant killing effects on CLL\u20101\u2009+\u2009CD123+ leukaemia cell lines and primary AML tumour cells. The killing efficiency of tandem CAR\u2010T cells in the case of single antigen expression is comparable to that of single target CAR\u2010T cells. When faced with dual target tumour cells, dual target CAR\u2010T cells significantly surpass single target CAR\u2010T cells. CD123\/CLL\u20101 CAR\u2010T cells in tandem targeted and killed CD123\u2010 and CLL\u20101\u2010positive leukaemia cell lines and released a large number of cytokines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>CD123\/CLL\u20101 CAR\u2010T cells in tandem can simultaneously target CD123 and CLL\u20101 on AML cells, demonstrating a significant ability to kill single antigens and multi\u2010target tumour cells. This suggests that CD123\/CLL\u20101 CAR\u2010T cells exhibit significant advantages in the expression of multiple antigens in a wide range of target cells, which may help overcome the challenges posed by tumour heterogeneity and evasion mechanisms.<\/jats:p><\/jats:sec>","DOI":"10.1111\/ejh.14104","type":"journal-article","created":{"date-parts":[[2023,9,15]],"date-time":"2023-09-15T12:33:19Z","timestamp":1694781199000},"page":"83-93","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Tandem bispecific <scp>CD123<\/scp>\/<scp>CLL<\/scp>\u20101 <scp>CAR\u2010T<\/scp> cells exhibit specific cytolytic effector functions against human acute myeloid leukaemia"],"prefix":"10.1111","volume":"112","author":[{"given":"Xiang\u2010Yu","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Hematology Huai'an Hospital Affiliated to Xuzhou Medical University, Huai'an Second People's Hospital  Huai'an 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