Bibliographic Details
| Title: |
Monitoring of Circulating CAR T Cells: Validation of a Flow Cytometric Assay, Cellular Kinetics, and Phenotype Analysis Following Tisagenlecleucel |
| Authors: |
Andreas Peinelt, Melanie Bremm, Hermann Kreyenberg, Claudia Cappel, Julia Banisharif-Dehkordi, Stephanie Erben, Eva Rettinger, Andrea Jarisch, Roland Meisel, Paul-Gerhardt Schlegel, Olaf Beck, Gesine Bug, Jan-Henning Klusmann, Thomas Klingebiel, Sabine Huenecke, Peter Bader |
| Source: |
Frontiers in Immunology, Vol 13 (2022) |
| Publisher Information: |
Frontiers Media S.A., 2022. |
| Publication Year: |
2022 |
| Collection: |
LCC:Immunologic diseases. Allergy |
| Subject Terms: |
acute lymphoblastic leukemia, chimeric antigen receptor (CAR), immunotherapy, flow cytometry, immune monitoring, Immunologic diseases. Allergy, RC581-607 |
| More Details: |
Chimeric antigen receptor (CAR) T cell therapy is a potent new treatment option for relapsed or refractory hematologic malignancies. As the monitoring of CAR T cell kinetics can provide insights into the activity of the therapy, appropriate CAR T cell detection methods are essential. Here, we report on the comprehensive validation of a flow cytometric assay for peripheral blood CD19 CAR T cell detection. Further, a retrospective analysis (n = 30) of CAR T cell and B cell levels over time has been performed, and CAR T cell phenotypes have been characterized. Serial dilution experiments demonstrated precise and linear quantification down to 0.05% of T cells or 22 CAR T cell events. The calculated detection limit at 13 events was confirmed with CAR T cell negative control samples. Inter-method comparison with real-time PCR showed appreciable correlation. Stability testing revealed diminished CAR T cell values already one day after sample collection. While we found long-term CAR T cell detectability and B cell aplasia in most patients (12/17), some patients (5/17) experienced B cell recovery. In three of these patients the coexistence of CAR T cells and regenerating B cells was observed. Repeat CAR T cell infusions led to detectable but limited re-expansions. Comparison of CAR T cell subsets with their counterparts among all T cells showed a significantly higher percentage of effector memory T cells and a significantly lower percentage of naïve T cells and T EMRA cells among CAR T cells. In conclusion, flow cytometric CAR T cell detection is a reliable method to monitor CAR T cells if measurements start without delay and sufficient T cell counts are given. |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
1664-3224 |
| Relation: |
https://www.frontiersin.org/articles/10.3389/fimmu.2022.830773/full; https://doaj.org/toc/1664-3224 |
| DOI: |
10.3389/fimmu.2022.830773 |
| Access URL: |
https://doaj.org/article/79daa92e8f4244719dcde74aa8452d48 |
| Accession Number: |
edsdoj.79daa92e8f4244719dcde74aa8452d48 |
| Database: |
Directory of Open Access Journals |
