ArticleFrontiers in oncology2026
Clinical implementation of a one-step no-wash flow cytometry method allows for real-time monitoring of patients treated with autologous CAR-T cells.
Article in Frontiers in oncology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Background: The best way to monitor Chimeric Antigen Receptor (CAR)-T cells persistence and expansion Methods: CAR-T cell monitoring was implemented between 2021 and 2024 in patients treated with autologous CD19-directed CAR-T therapies at our institution with a classical two steps and wash flow cytometry method. Analytical validation included determination of detection and quantification limits, linearity, precision, and inter-laboratory reproducibility. In 2024, the classical method was considerably optimized to a one-step no-wash format to reduce manual handling and improve sensitivity. Clinical relevance was assessed in a cohort of 29 patients treated with axicabtagene ciloleucel, correlating CAR-T expansion metrics with clinical endpoints. Results: The optimized one-step no-wash assay markedly improved analytical sensitivity, achieving a limit of detection of 0.3 cells/µL and a lower limit of quantification of 1.0 cells/µL, versus 2.0 and 5.0 cells/µL, respectively, when using the previous two steps and wash protocol, while demonstrating a strong concordance (r² = 0.984). Inter-assay coefficients of variation remained below 9%, confirming maintained precision despite workflow simplification. In 29 patients treated with axicabtagene ciloleucel (axicel), peak CAR-T expansion significantly correlated with objective responses (p< 0.01) and occurrences of immune effector cell-associated neurotoxicity syndrome (ICANS) (p = 0.02), supporting the clinical relevance of flow cytometry-based CAR-T monitoring in routine practice. Conclusion: This one-step no-wash flow cytometry assay combines enhanced sensitivity with improved operational efficiency and provides clinically informative CAR-T cell monitoring in standard-of-care settings.
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