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Updated: Feb 1, 2026

Generation of Human Chimeric Antigen Receptor Regulatory T Cells
Published on: January 3, 2025
A quantitative method for measuring the transfection efficiency of CD19-directed chimeric antigen receptor in target
Na Fang1, Niannian Zhong1, Tingxuan Gu1
1Department of Biochemistry and Molecular Biology, Medical School, Henan University, Kaifeng, China.
Background:
Adoptive cell therapy (ACT) based on chimeric antigen receptors (CARs) expressed on the surface of T cells shows a remarkable clinical outcome, particularly for B-cell malignancies. However, toxicity and side effects of CD19-redirected CAR T cells have been observed concurrently in most cases due to cytokine release and tumor cell lysis. Therefore, strictly controlling the amount of valid T cells re-transfused to patients seems to be an important step in reducing toxicity and side effects of CAR T cells. Transfection efficiency via lentiviral particles varies widely in different cases.
Objectives:
The aim of this study was to accurately calculate and control the number of valid CAR T cells through ACT because the restriction antibiotics gene or the fluorescence gene are not suitable for tracking or screening for valid transfected T cells.
Material And Methods:
We expressed and purified a GFP-CD19 fusion protein as a probe to measure the expression efficiency of CD19-redirected CAR on the cell surface in adherent and suspension cell lines.
Results:
We can precisely calculate the transfected efficiency of lentiviral particles by counting the number of GFP-labeled cells under a microscope, as well as calculate the percentage by comparing the number of GFP-labeled cells to total cells.
Conclusions:
We propose a method to control the number of valid cells in ACT and to reduce toxicity and side effects in clinical use - a convenient technique for monitoring the dosage of CAR T cells for patients.
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