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Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies

Cancer Biology & Medicine 2021年第4期18卷 1066-1079页

作者： Lu Han Jian Zhou Linlin Li Keshu Zhou Lingdi Zhao Xinghu Zhu Qingsong Yin Yufu Li Hongqin You Jishuai Zhang Yongping Song Quanli Gao Department of Immunology Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer HospitalZhengzhou 450008China Department of Hematology Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer HospitalZhengzhou 450008China Department of Medical Microbiology School of Basic Medical SciencesXinxiang Medical UniversityXinxiang 453003China The Shenzhen Pregene Biopharma Company Ltd.Shenzhen 518118China

Objective:Chimeric antigen receptor-modified T(CAR-T)cells have shown impressive results against relapsed/refractory B cell ***,the traditional manufacture of CAR-T cells requires leukapheresis to isolate large amounts of peripheral blood T cells,thus making some patients ineligible for the ***:We developed a simple method for CAR-T cell preparation requiring small volumes of peripheral ***,CD3+T cells isolated from 50 mL peripheral blood from patients(B-cell malignancies)were stimulated with immobilized anti-CD3/RetroNectin in 6-well plates and then transduced with CAR-expressing lentiviral *** 4 d,the T cells were transferred to culture bags for large-scale CAR-T cell *** vitro and animal experiments were performed to evaluate the activity of the manufactured CAR-T ***,29 patients with B-cell acute lymphoblastic leukemia(B-ALL)and 9 patients with B-cell lymphoma were treated with the CAR-T ***:The CAR-T cells were expanded to 1–3×10^(8) cells in 8–10 d and successfully killed B cell-derived malignant tumor cells in vitro and in *** patients with B-ALL,the complete remission rate was 93%1 month after CAR-T cell infusion;after 12 months,the overall survival(OS)and leukemia-free survival rates were 69%and 31%,*** patients with lymphoma,the objective response rate(including complete and partial remission)was 78%2 months after CAR-T cell infusion,and after 12 months,the OS and progression-free survival rates were 71%and 43%,***-release syndrome(CRS)occurred in 65.51%and 55.56%of patients with B-ALL and B-cell lymphoma,respectively;severe CRS developed in 20.69%of patients with B-ALL and in no patients with ***:Our novel method can generate sufficient numbers of CAR-T cells for clinical use from 50–100 mL peripheral blood,thus providing an alternative means of CAR-T cell generation for patients ineligible for leukapheresis.

关键词： fewer initial lymphocytes peripheral blood CAR-T cells B-cell malignancy acute lymphoblastic leukemia

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