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Published on: August 16, 2019
Development of antigen multimers for detection and evaluation of CAR T cells
Rasmus U W Friis1, Maria Ormhøj1, Cecilie S Krüger-Jensen1
1Department of Health Technology, Technical University of Denmark, Kgs. Lyngby, Denmark.
Background:
Chimeric antigen receptor (CAR) T cell therapy has transformed the treatment landscape of hematologic cancers by engineering T cells to specifically target and destroy cancer cells. Monitoring CAR T cell activity and function is essential for optimizing therapeutic outcomes, but existing tools for CAR detection are often limited in specificity and functional assessment capability.
Methods:
We developed dextran multimers by conjugating multiple CAR-specific antigens to a dextran backbone. The multimers were compared to previously reported antigen tetramers for their ability to stain and detect CAR T cells. Because these multimers incorporate the CAR target antigen, they uniquely enable assessment of CAR T cell functionality. We tested the staining and functional properties of the multimers across a range of CAR constructs with different affinities, using flow cytometry and microscopy.
Results:
The dextran multimers demonstrated high specificity and sensitivity in staining CAR T cells, with adjustable antigen density to optimize binding. Dextran multimers also enabled effective clustering and subsequent activation of CARs, showing their utility as both a staining and functional assessment tool. The multimers revealed that CARs with different affinities and clustering tendencies displayed varied binding and activation in response to different antigen densities.
Conclusion:
Dextran multimers offer a dual advantage as versatile reagents for both staining and functional analysis of CAR T cells. Their capacity to engage CARs with the specific antigen provides a valuable platform for evaluating CAR functionality, informing CAR design improvements, and enhancing therapeutic precision.
Insights
New dextran multimers effectively detect and assess chimeric antigen receptor (CAR) T cell function. These versatile tools improve CAR T cell therapy monitoring and CAR design for enhanced precision.
Area of Science:
- Immunology
- Biotechnology
- Cancer Therapy
Background:
- Chimeric antigen receptor (CAR) T cell therapy is a breakthrough in treating hematologic cancers.
- Accurate monitoring of CAR T cell activity is crucial for treatment optimization.
- Current CAR detection methods lack specificity and functional assessment capabilities.
Purpose of the Study:
- To develop and evaluate novel dextran multimers for enhanced CAR T cell detection and functional analysis.
- To compare the efficacy of dextran multimers against existing antigen tetramers.
- To assess the utility of multimers in evaluating CAR T cell functionality across various CAR constructs.
Main Methods:
- Conjugation of multiple CAR-specific antigens to a dextran backbone to create dextran multimers.
- Comparison of dextran multimers and antigen tetramers for CAR T cell staining and detection.
- Assessment of CAR T cell functionality using flow cytometry and microscopy with dextran multimers.
- Testing across diverse CAR constructs with varying affinities and antigen densities.
Main Results:
- Dextran multimers exhibit high specificity and sensitivity in staining CAR T cells.
- Adjustable antigen density on multimers optimizes CAR T cell binding.
- Multimers facilitate CAR clustering and activation, enabling functional assessment.
- CAR affinity and clustering influence binding and activation responses to antigen density.
Conclusions:
- Dextran multimers serve as versatile reagents for both CAR T cell staining and functional analysis.
- These multimers provide a platform for evaluating CAR functionality and informing therapeutic strategies.
- The developed multimers enhance precision in CAR T cell therapy and CAR design.
