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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Lu Han, Jian Zhou, Linlin Li, Keshu Zhou, Lingdi Zhao, Xinghu Zhu, Qingsong Yin, Yufu Li, Hongqin You, Jishuai Zhang, Yongping Song, Quanli Gao
Formato:	article
Lenguaje:	EN
Publicado:	China Anti-Cancer Association 2021
Materias:	fewer initial lymphocytes peripheral blood car-t cells b-cell malignancy acute lymphoblastic leukemia Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
Acceso en línea:	https://doaj.org/article/b31ae4dc1bc947628b4a687150159dd3
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:b31ae4dc1bc947628b4a687150159dd3
record_format	dspace
spelling	oai:doaj.org-article:b31ae4dc1bc947628b4a687150159dd32021-11-30T11:27:44ZCulturing adequate CAR-T cells from less peripheral blood to treat B-cell malignancies2095-394110.20892/j.issn.2095-3941.2021.0040https://doaj.org/article/b31ae4dc1bc947628b4a687150159dd32021-11-01T00:00:00Zhttp://www.cancerbiomed.org/index.php/cocr/article/view/1890https://doaj.org/toc/2095-3941Objective: 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.Lu HanJian ZhouLinlin LiKeshu ZhouLingdi ZhaoXinghu ZhuQingsong YinYufu LiHongqin YouJishuai ZhangYongping SongQuanli GaoChina Anti-Cancer Associationarticlefewer initial lymphocytesperipheral bloodcar-t cellsb-cell malignancyacute lymphoblastic leukemiaNeoplasms. Tumors. Oncology. Including cancer and carcinogensRC254-282ENCancer Biology & Medicine, Vol 18, Iss 4, Pp 1066-1079 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	fewer initial lymphocytes peripheral blood car-t cells b-cell malignancy acute lymphoblastic leukemia Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
spellingShingle	fewer initial lymphocytes peripheral blood car-t cells b-cell malignancy acute lymphoblastic leukemia Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282 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
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.
format	article
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
title	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_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_sort	culturing adequate car-t cells from less peripheral blood to treat b-cell malignancies
publisher	China Anti-Cancer Association
publishDate	2021
url	https://doaj.org/article/b31ae4dc1bc947628b4a687150159dd3
work_keys_str_mv	AT luhan culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT jianzhou culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT linlinli culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT keshuzhou culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT lingdizhao culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT xinghuzhu culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT qingsongyin culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT yufuli culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT hongqinyou culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT jishuaizhang culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT yongpingsong culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies AT quanligao culturingadequatecartcellsfromlessperipheralbloodtotreatbcellmalignancies
_version_	1718406645029011456

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

Ejemplares similares