Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies
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 inel...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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China Anti-Cancer Association
2021-11-01
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| Series: | Cancer Biology & Medicine |
| Subjects: | |
| Online Access: | http://www.cancerbiomed.org/index.php/cocr/article/view/1890 |
| _version_ | 1818833849304481792 |
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| author | Lu Han Jian Zhou Linlin Li Keshu Zhou Lingdi Zhao Xinghu Zhu Qingsong Yin Yufu Li Hongqin You Jishuai Zhang Yongping Song Quanli Gao |
| author_facet | Lu Han Jian Zhou Linlin Li Keshu Zhou Lingdi Zhao Xinghu Zhu Qingsong Yin Yufu Li Hongqin You Jishuai Zhang Yongping Song Quanli Gao |
| author_sort | Lu Han |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-19T02:25:27Z |
| format | Article |
| id | doaj.art-b31ae4dc1bc947628b4a687150159dd3 |
| institution | Directory Open Access Journal |
| issn | 2095-3941 |
| language | English |
| last_indexed | 2024-12-19T02:25:27Z |
| publishDate | 2021-11-01 |
| publisher | China Anti-Cancer Association |
| record_format | Article |
| series | Cancer Biology & Medicine |
| spelling | doaj.art-b31ae4dc1bc947628b4a687150159dd32022-12-21T20:39:57ZengChina Anti-Cancer AssociationCancer Biology & Medicine2095-39412021-11-011841066107910.20892/j.issn.2095-3941.2021.0040Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignanciesLu Han0Jian Zhou1Linlin Li2Keshu Zhou3Lingdi Zhao4Xinghu Zhu5Qingsong Yin6Yufu Li7Hongqin You8Jishuai Zhang9Yongping Song10Quanli Gao11Department of Immunology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Hematology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Medical Microbiology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453003, ChinaDepartment of Hematology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Immunology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Hematology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Hematology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Hematology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Immunology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaThe Shenzhen Pregene Biopharma Company, Ltd., Shenzhen 518118, ChinaDepartment of Hematology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaDepartment of Immunology, Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, ChinaObjective: 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.http://www.cancerbiomed.org/index.php/cocr/article/view/1890fewer initial lymphocytesperipheral bloodcar-t cellsb-cell malignancyacute lymphoblastic leukemia |
| spellingShingle | Lu Han Jian Zhou Linlin Li Keshu Zhou Lingdi Zhao Xinghu Zhu Qingsong Yin Yufu Li Hongqin You Jishuai Zhang Yongping Song Quanli Gao Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies Cancer Biology & Medicine fewer initial lymphocytes peripheral blood car-t cells b-cell malignancy acute lymphoblastic leukemia |
| title | Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies |
| title_full | Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies |
| title_fullStr | Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies |
| title_full_unstemmed | Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies |
| title_short | Culturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies |
| title_sort | culturing adequate car t cells from less peripheral blood to treat b cell malignancies |
| topic | fewer initial lymphocytes peripheral blood car-t cells b-cell malignancy acute lymphoblastic leukemia |
| url | http://www.cancerbiomed.org/index.php/cocr/article/view/1890 |
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