PMID- 25879549 OWN - NLM STAT- MEDLINE DCOM- 20160126 LR - 20181113 IS - 1756-8722 (Electronic) IS - 1756-8722 (Linking) VI - 8 DP - 2015 Feb 28 TI - AntiCD3Fv fused to human interleukin-3 deletion variant redirected T cells against human acute myeloid leukemic stem cells. PG - 18 LID - 10.1186/s13045-015-0109-5 [doi] LID - 18 AB - BACKGROUND: Leukemic stem cells (LSCs) are frequently seen as a cause of treatment failure and relapse in patients with acute myeloid leukemia (AML). Thus, successful new therapeutic strategies for the treatment of AML should aim at eradicating LSCs. The identification of targets on the cell surface of LSCs is getting more and more attention. Among these, CD123, also known as the interleukin-3 (IL3)-receptor alpha chain, has been identified as a potential immunotherapeutic target due to its overexpression on LSCs in AML as well as on AML blasts, rather than normal hematopoietic stem cells. METHODS: We constructed a CD123-targeted fusion protein antiCD3Fv-⊿IL3, with one binding site for T cell antigen receptor (TCRCD3) and the other for CD123, by recombinant gene-engineering technology. Cysteine residues were introduced into the V domains of the antiCD3Fv segment to enhance its stability by locking the two chains of Fv together with disulfide covalent bonds. The stability and cytotoxicity of the two fusion proteins were detected in vitro and in vivo. RESULTS: Both fusion proteins were produced and purified from Escherichia coli 16C9 cells with excellent yields in fully active forms. High-binding capability was observed between these two fusion proteins and human IL3R, leading to the specific lysis of CD123-expressing cell lines KG1a; also, mononuclear cells from primary AML patients were inhibited in a colony forming assay in vitro, presumably by redirecting T lymphocytes in vitro. In addition, they displayed an antileukemic activity against KG1a xenografts in non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice, especially disulfide-stabilized (ds)-antiCD3Fv-⊿IL3 for its improved stability. CONCLUSIONS: These results suggest that both fusion proteins display the antileukemic activity against CD123-expressing cell lines as well as leukemic progenitors in vitro and in vivo, especially ds-antiCD3Fv-⊿IL3. They could be the promising candidates for future immunotherapy of AML. FAU - Fan, Dongmei AU - Fan D AD - State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China. fdm19691217@163.com. FAU - Li, Zhenzhen AU - Li Z AD - State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China. lizhenvvv@163.com. FAU - Zhang, Xiaolong AU - Zhang X AD - State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China. zhxlongmu@126.com. FAU - Yang, Yuqi AU - Yang Y AD - School of Pharmacy, Tianjin Medical University, Tianjin, 300070, People's Republic of China. yangyvjian@gmail.com. FAU - Yuan, Xiangfei AU - Yuan X AD - State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China. xfyuanca@sohu.com. FAU - Zhang, Xiuli AU - Zhang X AD - State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China. zxlics@yeah.net. FAU - Yang, Ming AU - Yang M AD - State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China. my1970@sina.com. FAU - Zhang, Yizhi AU - Zhang Y AD - Central Hospital of Karamay, Karamay, Xinjiang, 834000, China. yz5168813@sohu.com. FAU - Xiong, Dongsheng AU - Xiong D AD - State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China. dsxiong@ihcams.ac.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150228 PL - England TA - J Hematol Oncol JT - Journal of hematology & oncology JID - 101468937 RN - 0 (CD3 Complex) RN - 0 (IL3RA protein, human) RN - 0 (Interleukin-3 Receptor alpha Subunit) RN - 0 (Recombinant Fusion Proteins) SB - IM MH - Animals MH - CD3 Complex/*immunology MH - Cell Line, Tumor MH - Female MH - Flow Cytometry MH - Humans MH - Immunoblotting MH - Immunotherapy/*methods MH - Interleukin-3 Receptor alpha Subunit/*immunology MH - Leukemia, Myeloid, Acute/*immunology/pathology MH - Mice MH - Mice, Inbred NOD MH - Mice, SCID MH - Neoplastic Stem Cells/*immunology/pathology MH - Recombinant Fusion Proteins/chemical synthesis/*pharmacology MH - T-Lymphocytes/immunology MH - Xenograft Model Antitumor Assays PMC - PMC4389834 EDAT- 2015/04/17 06:00 MHDA- 2016/01/27 06:00 PMCR- 2015/02/28 CRDT- 2015/04/17 06:00 PHST- 2014/10/02 00:00 [received] PHST- 2015/01/13 00:00 [accepted] PHST- 2015/04/17 06:00 [entrez] PHST- 2015/04/17 06:00 [pubmed] PHST- 2016/01/27 06:00 [medline] PHST- 2015/02/28 00:00 [pmc-release] AID - 10.1186/s13045-015-0109-5 [pii] AID - 109 [pii] AID - 10.1186/s13045-015-0109-5 [doi] PST - epublish SO - J Hematol Oncol. 2015 Feb 28;8:18. doi: 10.1186/s13045-015-0109-5.