嵌合抗原受体
CD19
流式细胞术
免疫系统
细胞毒性T细胞
颗粒酶B
B细胞
医学
免疫疗法
癌症研究
T细胞
抗体
免疫学
生物
体外
生物化学
作者
Ulrich Blache,Reinhold Weiß,Andreas Boldt,Michael Kapinsky,André‐René Blaudszun,Andrea Quaiser,Annabelle Pohl,Tewfik Miloud,Mégane Burgaud,Vladan Vučinić,Uwe Platzbecker,Ulrich Sack,Frank Stephan,Ulrike Koehl
标识
DOI:10.3389/fimmu.2021.658314
摘要
Adoptive immunotherapy using chimeric antigen receptor (CAR)-T cells has achieved successful remissions in refractory B-cell leukemia and B-cell lymphomas. In order to estimate both success and severe side effects of CAR-T cell therapies, longitudinal monitoring of the patient’s immune system including CAR-T cells is desirable to accompany clinical staging. To conduct research on the fate and immunological impact of infused CAR-T cells, we established standardized 13-colour/15-parameter flow cytometry assays that are suitable to characterize immune cell subpopulations in the peripheral blood during CAR-T cell treatment. The respective staining technology is based on pre-formulated dry antibody panels in a uniform format. Additionally, further antibodies of choice can be added to address specific clinical or research questions. We designed panels for the anti-CD19 CAR-T therapy and, as a proof of concept, we assessed a healthy individual and three B-cell lymphoma patients treated with anti-CD19 CAR-T cells. We analyzed the presence of anti-CD19 CAR-T cells as well as residual CD19+ B cells, the activation status of the T-cell compartment, the expression of co-stimulatory signaling molecules and cytotoxic agents such as perforin and granzyme B. In summary, this work introduces standardized and modular flow cytometry assays for CAR-T cell clinical research, which could also be adapted in the future as quality controls during the CAR-T cell manufacturing process.
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