Image_1_Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications.jpeg
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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References
- https://doi.org//10.1038/s41577-020-0306-5
- https://doi.org//10.1056/NEJMoa1709866
- https://doi.org//10.1056/NEJMoa1707447
- https://doi.org//10.1056/NEJMoa1804980
- https://doi.org//10.1056/NEJMoa1914347
- https://doi.org//10.1038/d41573-019-00090-z
- https://doi.org//10.1038/s41591-020-1081-3
- https://doi.org//10.1038/mto.2016.11
- https://doi.org//10.1038/nrclinonc.2017.148
- https://doi.org//10.1016/j.bbmt.2018.12.758
- https://doi.org//10.1158/2159-8290.CD-16-0040
- https://doi.org//10.1038/s41409-019-0602-5
- https://doi.org//10.1002/eji.201970107
- https://doi.org//10.1080/14737159.2019.1691918
- https://doi.org//10.1002/cyto.b.21833
- https://doi.org//10.1038/nri3158
- https://doi.org//10.1002/cyto.a.20623
- https://doi.org//10.1038/s41467-019-13880-1
- https://doi.org//10.1038/s41591-018-0010-1
- https://doi.org//10.1056/NEJMoa1407222
- https://doi.org//10.1016/j.omtm.2019.12.018
- https://doi.org//10.3390/cancers12071970
- https://doi.org//10.1182/bloodadvances.2019000525
- https://doi.org//10.1016/j.jcyt.2020.01.008
- https://doi.org//10.1016/j.ymthe.2016.10.020
- https://doi.org//10.1016/S1470-2045(18)30864-7
- https://doi.org//10.3389/fimmu.2020.01770
- https://doi.org//10.1038/s41591-020-1061-7
- https://doi.org//10.1002/cyto.b.21880
- https://doi.org//10.1038/s41591-018-0201-9
- https://doi.org//10.1182/blood-2019-128745
- https://doi.org//10.1016/j.jcyt.2020.04.070
- https://doi.org//10.1002/cyto.b.21622
- https://doi.org//10.1002/cyto.a.23570
- https://doi.org//10.1155/2020/1938704
- https://doi.org//10.1038/s41598-019-39631-2
- https://doi.org//10.1016/j.ccell.2020.02.001
- https://doi.org//10.3389/fimmu.2018.00283
- https://doi.org//10.1038/s41592-020-0862-4
- https://doi.org//10.1038/s41587-020-0462-y
- https://doi.org//10.1038/d41586-020-02675-w
- https://doi.org//10.1089/hum.2017.254
- https://doi.org//10.1089/hum.2017.157
- https://doi.org//10.1089/hgtb.2019.039
- https://doi.org//10.1089/hum.2017.111
- https://doi.org//10.1159/000495772
- https://doi.org//10.1016/j.jcyt.2016.05.009
- https://doi.org//10.1016/j.jcyt.2017.09.005
- https://doi.org//10.1172/jci.insight.121867
- https://doi.org//10.1002/cyto.a.23852
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Categories
- Transplantation Immunology
- Tumour Immunology
- Immunology not elsewhere classified
- Immunology
- Veterinary Immunology
- Animal Immunology
- Genetic Immunology
- Applied Immunology (incl. Antibody Engineering, Xenotransplantation and T-cell Therapies)
- Autoimmunity
- Cellular Immunology
- Humoural Immunology and Immunochemistry
- Immunogenetics (incl. Genetic Immunology)
- Innate Immunity