An Ex Vivo 3D Tumor Microenvironment-Mimicry Culture to Study TAM Modulation of Cancer Immunotherapy
Abstract
:1. Introduction
2. Materials and Methods
2.1. Antibodies and Flow Cytometry
2.2. Enzyme-Linked Immunosorbent Cytokine Assays (ELISA)
2.3. Lentiviral Vectors
2.4. Human Monocyte-Derived Macrophage (MDM) Culture and Polarization
2.5. Generation of Human BCMA CAR-Engineered T (BCAR-T) Cells, Mesothelin CAR-Engineered T (MCAR-T) Cells, and NY-ESO-1-Specific TCR-Engineered T (ESO-T) Cells
2.6. In Vitro Mixed Mφ/T-Cell Reaction Assay
2.7. In Vitro Tumor Cell Killing Assay
2.8. Ex Vivo 3D TME-Mimicry Culture
3. Results
3.1. Generation of Human Monocyte-Derived M2-Polarized Macrophages
3.2. Validation of M2-Polarized Macrophage Immunosuppressive Function
3.3. Validation of PD-1/PD-L1 Blockade on Antagonizing M2 Macrophage-Mediated Immunosuppression
3.4. Validation of MAO-A Blockade on Antagonizing M2 Macrophage-Mediated Immunosuppression
3.5. Development of an Ex Vivo 3D TME-Mimicry Culture to Study TAM Modulation of T-Cell Antitumor Reactivity
3.6. PD-1/PD-L1 Blockade Antagonizes TAM-Suppression of T-Cell Antitumor Reactivity
3.7. MAO-A Blockade Antagonizes TAM-Suppression of T-Cell Antitumor Reactivity
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Li, Y.-R.; Yu, Y.; Kramer, A.; Hon, R.; Wilson, M.; Brown, J.; Yang, L. An Ex Vivo 3D Tumor Microenvironment-Mimicry Culture to Study TAM Modulation of Cancer Immunotherapy. Cells 2022, 11, 1583. https://doi.org/10.3390/cells11091583
Li Y-R, Yu Y, Kramer A, Hon R, Wilson M, Brown J, Yang L. An Ex Vivo 3D Tumor Microenvironment-Mimicry Culture to Study TAM Modulation of Cancer Immunotherapy. Cells. 2022; 11(9):1583. https://doi.org/10.3390/cells11091583
Chicago/Turabian StyleLi, Yan-Ruide, Yanqi Yu, Adam Kramer, Ryan Hon, Matthew Wilson, James Brown, and Lili Yang. 2022. "An Ex Vivo 3D Tumor Microenvironment-Mimicry Culture to Study TAM Modulation of Cancer Immunotherapy" Cells 11, no. 9: 1583. https://doi.org/10.3390/cells11091583