ArticleCytometry. Part A : the journal of the International Society for Analytical Cytology2023
Monitoring anti-CD19 chimeric antigen receptor T cell population by flow cytometry and its consistency with digital droplet polymerase chain reaction.
Article in Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- CAR T-cell Kinetics, Persistence, and Clinical Outcomes in Adult Patients with Relapsed/Refractory B-cell ALL Treated with Obecabtagene Autoleucel in the FELIX Study.Cancer research communications · 2026Trial
- Universal high-sensitivity CAR T-cell monitoring by targeting linker sequences.Frontiers in immunology · 2026Article
- A universally applicable toolbox for single-molecule quantification of chimeric antigen receptors using linker-resolvedFrontiers in immunology · 2026Article
- Clinical implementation of a one-step no-wash flow cytometry method allows for real-time monitoring of patients treated with autologous CAR-T cells.Frontiers in oncology · 2026Article
- Current Insights of Post-Infusion CAR T Expansion and Persistence for Large B-Cell Lymphoma.Cancers · 2025Review
- Characterization of a multiplex digital PCR assay to quantify total T cells relative to chimeric antigen receptor-positive T cells.Molecular therapy. Oncology · 2025Article
- Review
- CAR-Aptamers Enable Traceless Enrichment and Monitoring of CAR-Positive Cells and Overcome Tumor Immune Escape.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Unlocking Predictive Power: Quantitative Assessment of CAR-T Expansion with Digital Droplet Polymerase Chain Reaction (ddPCR).International journal of molecular sciences · 2024Article
- Long-term analysis of cellular immunity in patients with RRMM treated with CAR-T cell therapy.Clinical and experimental medicine · 2023Article
- Early quantification of anti-CD19 CAR T cells by flow cytometry predicts response in R/R DLBCL.Blood advances · 2023Article
- The Evolving Landscape of Potency Assays.Advances in experimental medicine and biology · 2023Article
- Research Topic: Measurable Residual Disease in Hematologic Malignancies. Can digital droplet PCR improve measurable residual disease monitoring in chronic lymphoid malignancies?Frontiers in oncology · 2023Review
- A new story for an old challenge: Would flow cytometry beat molecular biology in monitoring chimeric antigen receptor T cell pharmacokinetics?Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2023Article
- Monitoring anti-CD19 chimeric antigen receptor T cell population by flow cytometry and its consistency with digital droplet polymerase chain reaction.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2023Article
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7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Anti-CD19 chimeric antigen receptor (CAR19) T cell therapy has produced impressive clinical efficacy in patients with relapsed or refractory B-cell malignancies. As a living drug, monitoring the pharmacokinetics of CAR T cells in vivo is an important part of clinical work, which provides valuable information for assessing therapeutic response and related side effects. However, no guidelines are available regarding the detection and quantification of CAR T cells. Flow cytometry is a convenient and commonly used method in monitoring CAR T cell kinetics, but its performance remains to be validated. By using a commercial anti-idiotype antibody that detects unique epitopes on the most popular CAR19 construct, we evaluated important performance parameters, including specificity, lower limit of detection, lower limit of quantification, and precision of flow cytometry in the detection and quantification of CAR19 T cells. Consistency between the results generated by flow cytometry and droplet digital PCR was then investigated in 188 pairs of clinical data and in cell line experiments. Rabbit anti-mouse FMC63 monoclonal antibody possesses high specificity in the detection of CAR19 positive cells by FCM with a cut-off value of 0.05%. The results produced by flow cytometry and ddPCR were well correlated in the clinical samples and in cell lines, but the correlation deteriorated as the abundance of CAR19 positive cells decreased. This was especially evident with less than 0.5% of lymphocytes in clinical data, possibly due to reduced precision (indicated by intra- and inter-assay coefficients of variability) of both droplet digital PCR and flow cytometry. We demonstrated that flow cytometry using anti-idiotype antibody is a reliable and robust approach in the detection and quantification of CAR19 T cells in vivo and has good consistency with droplet digital PCR in monitoring CAR19 T cell kinetics.
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