Novel Bi-Specific Immuno-Modulatory Tribodies Potentiate T Cell Activation and Increase Anti-Tumor Efficacy
Abstract
:1. Introduction
2. Results
2.1. In Vitro Effects of Combined Immunomodulatory mAbs on the Stimulation of hPBMCs
2.2. Construction of Novel Immunomodulatory Tribodies
2.3. Purification and Analysis of the Stability of the Tribodies
2.4. Binding of the Novel Tribodies to Their Targets
2.5. Evaluation of Tribody Interference in the Binding of ICs with Their Ligands
2.6. Effects of the Tribodies on the Activation of hPBMCs
2.7. In Vitro Antitumor Effects of the Tribodies on Breast Cancer Cells
2.8. In Vitro Cytotoxic Effects of Tribodies on Breast Tumor Cells Co-Cultured with Human Lymphocytes
2.9. Analysis of Possible Side Effects of TRs on Circulating Naive Lymphocytes
2.10. Comparison of the Cardiotoxic Side Effects of Novel Tribodies with Those of the Respective Parental Compounds Used in Combination
3. Discussion
4. Materials and Methods
4.1. Cell Cultures
4.2. Antibodies and Human Recombinant Proteins
4.3. Production and Purification of Tribodies
4.4. Cytotoxicity and Cell Viability Assays
4.5. Cytokine Secretion Assays
4.6. ELISA Assays
4.6.1. Binding Assays
4.6.2. Competitive Assays
4.7. Western Blotting Analysis of Cell Extracts
4.8. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Protein ID | Sequence |
---|---|
TR0102 | (Fab)LAG-3-(scFv)2xPD-L1 |
TR0304 | (Fab)PD-L1-(scFv)2xLAG-3 |
TR0506 | (Fab)LAG-3-(scFv)2xPD-1 |
TR0708 | (Fab)PD-1-(scFv)2xLAG-3 |
Kd | TR0102 | TR0304 | TR0506 | TR0708 |
---|---|---|---|---|
hPBMCs | 2.265 nM | 1.910 nM | 3.439 nM | 1.057 nM |
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Passariello, M.; Yoshioka, A.; Takahashi, K.; Hashimoto, S.-i.; Rapuano Lembo, R.; Manna, L.; Nakamura, K.; De Lorenzo, C. Novel Bi-Specific Immuno-Modulatory Tribodies Potentiate T Cell Activation and Increase Anti-Tumor Efficacy. Int. J. Mol. Sci. 2022, 23, 3466. https://doi.org/10.3390/ijms23073466
Passariello M, Yoshioka A, Takahashi K, Hashimoto S-i, Rapuano Lembo R, Manna L, Nakamura K, De Lorenzo C. Novel Bi-Specific Immuno-Modulatory Tribodies Potentiate T Cell Activation and Increase Anti-Tumor Efficacy. International Journal of Molecular Sciences. 2022; 23(7):3466. https://doi.org/10.3390/ijms23073466
Chicago/Turabian StylePassariello, Margherita, Asami Yoshioka, Kota Takahashi, Shu-ichi Hashimoto, Rosa Rapuano Lembo, Lorenzo Manna, Koji Nakamura, and Claudia De Lorenzo. 2022. "Novel Bi-Specific Immuno-Modulatory Tribodies Potentiate T Cell Activation and Increase Anti-Tumor Efficacy" International Journal of Molecular Sciences 23, no. 7: 3466. https://doi.org/10.3390/ijms23073466