CAR-T细胞疗法的自动化设备及展望
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摘要
嵌合抗原受体(chimeric antigen receptor,CAR)T细胞疗法是近年来发展非常迅速的一种细胞治疗技术。通过从患者或者捐献者外周血分离获得T细胞,并进一步激活、基因修饰、扩增从而获得CAR-T细胞,洗涤浓缩后回输患者体内进行治疗。修饰后的T细胞肿瘤靶向性、杀伤活性更强,在血液肿瘤疾病临床试验中疗效明显,在实体瘤的临床试验中也崭露头角。CAR-T细胞疗法整个过程的安全性、无菌性、纯度等质量控制极其重要。确保CAR-T细胞疗法生产过程的标准化、封闭化与自动化,做到药效和风险可控是首要任务。对CAR-T细胞疗法工艺与自动化设备以及面临的工程化问题进行综述,以期为CAR-T细胞疗法工程化提供参考。
Chimeric antigen receptor(CAR)-T cell therapy has been a rapidly developing cell therapy technique in recent years. CAR-T cells are generated from the T cells of patients' or donations' peripheral blood, followed with activation, genetically modification and expansion. CAR-T cells are washed and concentrated prior to reinfuse into the patients. The modifi ed T cell with stronger tumor-orientated targeting ability and killing activity has achieved a remarkable effect on the treatment of hematologic tumors and clinical trials of solid tumors. Quality control on the safety, sterility and purity throughout the entire protocol is absolutely crucial. Therefore, the primary task is to ensure the standardization, closing and automation of CAR-T product process. This paper mainly reviewed the process and automatic devices for CAR-T and the problem for engineering, which could provide a reference for the CAR-T cell therapy engineering.
Chimeric antigen receptor(CAR)-T cell therapy has been a rapidly developing cell therapy technique in recent years. CAR-T cells are generated from the T cells of patients' or donations' peripheral blood, followed with activation, genetically modification and expansion. CAR-T cells are washed and concentrated prior to reinfuse into the patients. The modifi ed T cell with stronger tumor-orientated targeting ability and killing activity has achieved a remarkable effect on the treatment of hematologic tumors and clinical trials of solid tumors. Quality control on the safety, sterility and purity throughout the entire protocol is absolutely crucial. Therefore, the primary task is to ensure the standardization, closing and automation of CAR-T product process. This paper mainly reviewed the process and automatic devices for CAR-T and the problem for engineering, which could provide a reference for the CAR-T cell therapy engineering.
引文
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[4]李欢欢,朱平,伍学强,等.应用CD19修饰的嵌合抗原受体T细胞治疗淋巴细胞白血病[J].中国实验血液学杂志,2014,22(6):1753-1756.
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[13]刘炳峰.基于VRC01广谱中和抗体的CAR-T细胞特异性清除再激活的HIV-1潜伏感染细胞[D].广州:中山大学,2016.
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[15]骆群,黎燕.CD3、CD28抗体增强淋巴细胞转化功能的研究[J].军医进修学院学报,2005,26(6):479-480.
[16]LEVINE B.Personalized cell-based medicine:activated and expanded T cells for adoptive immunotherapy[J].Bio Processing,2007,6:14-19.
[17]KAARTINEN T,LUOSTARINEN A,MALINIEMI P,et al.Low interleukin-2 concentration favors generation of early memory T cells over effector phenotypes during chimeric antigen receptor T-cell expansion[J].Cytotherapy,2017,19(6):689-702.
[18]LEVINE B L,MISKIN J,WONNACOTT K,et al.Global manufact uring of CAR T cell therapy[J].Molecular Therapy:Methods&Clinical Development,2017,4:92-101.
[19]ZHANG C,LIU J,ZHONG J F,et al.Engineering CAR-T cells[J].Biomarker Research,2017,5:22.
[20]王强力.靶向CD19的嵌合抗原受体修饰的T细胞(CD19-CAR-T)的构建及其对CD19阳性血液肿瘤细胞的杀伤研究[D].苏州:苏州大学,2016.
[21]HOLLYMAN D,STEFANSKI J,PRZYBYLOWSKI M,et al.Manufacturing validation of biologically functional T cells targeted to CD19 antigen for autologous adoptive cell therapy[J].Journal of Immunotherapy,2009,32(2):169-180.
[22]SOMERVILLE R P,DEVILLIER L,PARKHURST M R,et al.Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE?bioreactor[J].Journal of Translational Medicine,2012,10:69.
[23]VAVROVA K,VRABCOVA P,FILIPP D,et al.Generation of T cell effectors using tumor cell-loaded dendritic cells for adoptive T cell therapy[J].Medical Oncology,2016,33:136.
[2]王艺,赵颖颖,韩双印.基于嵌合抗原受体修饰T细胞的肿瘤免疫治疗新策略[J].中国肿瘤生物治疗杂志,2013,20(4):383-390.
[3]郑敏,张岚.CAR-T抗肿瘤研究的现状及展望[J].山东大学学报(医学版),2016,54(11):1-18.
[4]李欢欢,朱平,伍学强,等.应用CD19修饰的嵌合抗原受体T细胞治疗淋巴细胞白血病[J].中国实验血液学杂志,2014,22(6):1753-1756.
[5]OLWEUS J.Manufacture of CAR-T cells in the body[J].Nature Biotechnology,2017,35(6):520-521.
[6]尹德成.血细胞分离机成分采集过程控制及程序实现[D].哈尔滨:哈尔滨工业大学,2014.
[7]张晗.全自动血细胞分离机分离采集技术及离心机构设计[D].哈尔滨:哈尔滨工业大学,2013.
[8]王扬,唐古生,徐丽丽,等.利用Mig R1质粒构建CD19-CAR逆转录病毒载体优化其对人原代T细胞的转染效率[J].中华血液学杂志,2015,36(4):331-336.
[9]王云飞,王艺,王春荣,等.CD3/CD28免疫磁珠法体外扩增外周血T淋巴细胞[J].细胞与分子免疫学杂志,2014,30(10):1090-1092.
[10]GARDNER R A,FINNEY O,ANNESLEY C,et al.Intent-to-treat leukemia remission by CD19 CAR T cells of definedformulation and dose in children and young adults[J].Blood,2017,129(25):3322-3331.
[11]MAUS M V,FRAIETTA J A,LEVINE B L,et al.Adoptive immunotherapy for cancer or viruses[J].Immunological Reviews,2014,257(1):14-38.
[12]BRECTJENS R J,SANTOS E,NIKHAMIN Y,et al.Genetically targeted T cells eradicate systemic acute lymphoblastic leukemia xenografts[J].Cancer Therapy:Preclinical,2007,13(18):5426-5435.
[13]刘炳峰.基于VRC01广谱中和抗体的CAR-T细胞特异性清除再激活的HIV-1潜伏感染细胞[D].广州:中山大学,2016.
[14]LEVINE B.Performance-enhancing drugs:design and production of redirected chimeric antigen receptor(CAR)T cells[J].Cancer Gene Therapy,2015,22(2):79-84.
[15]骆群,黎燕.CD3、CD28抗体增强淋巴细胞转化功能的研究[J].军医进修学院学报,2005,26(6):479-480.
[16]LEVINE B.Personalized cell-based medicine:activated and expanded T cells for adoptive immunotherapy[J].Bio Processing,2007,6:14-19.
[17]KAARTINEN T,LUOSTARINEN A,MALINIEMI P,et al.Low interleukin-2 concentration favors generation of early memory T cells over effector phenotypes during chimeric antigen receptor T-cell expansion[J].Cytotherapy,2017,19(6):689-702.
[18]LEVINE B L,MISKIN J,WONNACOTT K,et al.Global manufact uring of CAR T cell therapy[J].Molecular Therapy:Methods&Clinical Development,2017,4:92-101.
[19]ZHANG C,LIU J,ZHONG J F,et al.Engineering CAR-T cells[J].Biomarker Research,2017,5:22.
[20]王强力.靶向CD19的嵌合抗原受体修饰的T细胞(CD19-CAR-T)的构建及其对CD19阳性血液肿瘤细胞的杀伤研究[D].苏州:苏州大学,2016.
[21]HOLLYMAN D,STEFANSKI J,PRZYBYLOWSKI M,et al.Manufacturing validation of biologically functional T cells targeted to CD19 antigen for autologous adoptive cell therapy[J].Journal of Immunotherapy,2009,32(2):169-180.
[22]SOMERVILLE R P,DEVILLIER L,PARKHURST M R,et al.Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE?bioreactor[J].Journal of Translational Medicine,2012,10:69.
[23]VAVROVA K,VRABCOVA P,FILIPP D,et al.Generation of T cell effectors using tumor cell-loaded dendritic cells for adoptive T cell therapy[J].Medical Oncology,2016,33:136.