RT Journal Article
SR Electronic
T1 Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies
JF Cancer Biology & Medicine
JO Cancer Biol Med
FD China Anti-Cancer Association
SP 1066
OP 1079
DO 10.20892/j.issn.2095-3941.2021.0040
VO 18
IS 4
A1 Lu Han
A1 Jian Zhou
A1 Linlin Li
A1 Keshu Zhou
A1 Lingdi Zhao
A1 Xinghu Zhu
A1 Qingsong Yin
A1 Yufu Li
A1 Hongqin You
A1 Jishuai Zhang
A1 Yongping Song
A1 Quanli Gao
YR 2021
UL http://www.cancerbiomed.org/content/18/4/1066.abstract
AB Objective: Chimeric antigen receptor-modified T (CAR-T) cells have shown impressive results against relapsed/refractory B cell malignancies. However, 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 procedure.Methods: We developed a simple method for CAR-T cell preparation requiring small volumes of peripheral blood. First, 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 vector. After 4 d, the T cells were transferred to culture bags for large-scale CAR-T cell expansion. In vitro and animal experiments were performed to evaluate the activity of the manufactured CAR-T cells. Finally, 29 patients with B-cell acute lymphoblastic leukemia (B-ALL) and 9 patients with B-cell lymphoma were treated with the CAR-T cells.Results: The CAR-T cells were expanded to 1–3 × 108 cells in 8–10 d and successfully killed B cell-derived malignant tumor cells in vitro and in vivo. For 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%, respectively. For 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%, respectively. Cytokine-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 lymphoma.Conclusions: 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.