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 CART
cell manufacturing process.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:84294 |
| Date | 24 March 2023 |
| Creators | Blache, Ulrich, Weiss, Ronald, Boldt, Andreas, Kapinsky, Michael, Blaudszun, André-René, Quaiser, Andreas, Pohl, Annabelle, Miloud, Tewfik, Burgaud, Mégane, Vucinic, Vladan, Platzbecker, Uwe, Sack, Ulrich, Fricke, Stephan, Koehl, Ulrike |
| Publisher | Frontiers Research Foundation |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 1664-3224, 658314 |
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