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Updated: Jun 1, 2025
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Generation of Human Chimeric Antigen Receptor Regulatory T Cells
Published on: January 3, 2025
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Generation of Human Chimeric Antigen Receptor Regulatory T Cells
Russell W Cochrane1, Rob A Robino1, Leonardo M R Ferreira2
1Department of Microbiology and Immunology, Medical University of South Carolina; Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina; Hollings Cancer Center, Medical University of South Carolina.
Journal of Visualized Experiments : Jove
|January 20, 2025
Summary
This study details methods for creating engineered regulatory T cells (Tregs) using Chimeric Antigen Receptor (CAR) technology. These CAR Tregs offer potential for treating autoimmune diseases and transplant rejection by redirecting immune suppression.
Area of Science:
- Immunology
- Cell Therapy
- Cancer Immunotherapy
Background:
- Chimeric Antigen Receptor (CAR) T-cell therapy has shown significant success in treating hematological cancers.
- Regulatory T cells (Tregs) play a crucial role in immune regulation and inhibition.
- Adapting CAR technology to Tregs presents therapeutic opportunities for immune-mediated diseases.
Purpose of the Study:
- To describe methodologies for generating stable human CAR Tregs.
- To enable the redirection of Treg immunosuppressive activity for therapeutic applications.
- To provide insights into the behavior of CAR Tregs in preclinical and clinical settings.
Main Methods:
- Isolation of bona fide Tregs from human peripheral blood.
- Genetic modification of Tregs using lentivirus or CRISPR/Cas9-aided knock-in with AAV-mediated HDR.
- Ex vivo expansion and assessment of CAR Treg phenotypic stability and in vitro suppressive function.
Main Results:
- Detailed methodologies for Treg isolation and genetic modification are established.
- Stable human CAR Tregs can be generated and expanded ex vivo.
- Assessment of CAR Treg stability and function provides critical preclinical data.
Conclusions:
- The described methodologies facilitate the development of CAR Treg-based cell therapies.
- Engineered Tregs hold promise for treating autoimmune diseases, transplant rejection, and graft-versus-host disease.
- This work provides a foundation for future clinical applications of CAR Treg therapy.
