Adeno-Associated Virus (AAV)-Delivered Exosomal TAT and BiTE Molecule CD4-αCD3 Facilitate the Elimination of CD4 T Cells Harboring Latent HIV-1
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Transfection
2.2. Exosome Preparation
2.3. Molecular Cloning
2.4. Generation of CD4-αCD3
2.5. Immunoprecipitation
2.6. Western Blot
2.7. HIV-1 p24 Elisa Assay
2.8. Study Subjects
2.9. Study Approval
2.10. Isolation of Human T Cells
2.11. Isolation of Cytotoxic T Lymphocytes (CTLs)
2.12. Measurement of Intracellular HIV-1 mRNA
2.13. HIV Elimination (HIVE) Assay
2.14. Production of AAV Viruses
2.15. Infection of Balb/cJ Mice
2.16. Immunohistochemistry (IHC) and Hematoxylin and Eosin (H&E) Staining
2.17. Intact Proviral DNA Assay (IPDA)
2.18. Statistical Analysis
3. Results
3.1. Cytotoxic T Lymphocytes (CTLs) from Healthy Donor Blood Eliminate HIV-1-Infected Cells
3.2. Autologous CTLs Fail to Effectively Eliminate CD4+ T Cells with Latent HIV-1 Reactivated by Exo-Tat
3.3. Generation of a BiTE Molecule CD4-αCD3
3.4. CD4-αCD3 Mediates Killing of Cells Stably Expressing HIV-1
3.5. CD4-αCD3 Mediates Killing of Chronically HIV-1-Infected Primary CD4+ T Lymphocytes
3.6. Construction of an AAV Vector Encoding Exo-Tat, IL16lamp2b and CD4-αCD3
3.7. AAV-Eliminator Reduces Latent HIV-1 Reservoir in T Cells Isolated from PWH
4. Discussion
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Tang, X.; Lu, H.; Tarwater, P.M.; Silverberg, D.L.; Schorl, C.; Ramratnam, B. Adeno-Associated Virus (AAV)-Delivered Exosomal TAT and BiTE Molecule CD4-αCD3 Facilitate the Elimination of CD4 T Cells Harboring Latent HIV-1. Microorganisms 2024, 12, 1707. https://doi.org/10.3390/microorganisms12081707
Tang X, Lu H, Tarwater PM, Silverberg DL, Schorl C, Ramratnam B. Adeno-Associated Virus (AAV)-Delivered Exosomal TAT and BiTE Molecule CD4-αCD3 Facilitate the Elimination of CD4 T Cells Harboring Latent HIV-1. Microorganisms. 2024; 12(8):1707. https://doi.org/10.3390/microorganisms12081707
Chicago/Turabian StyleTang, Xiaoli, Huafei Lu, Patrick M. Tarwater, David L. Silverberg, Christoph Schorl, and Bharat Ramratnam. 2024. "Adeno-Associated Virus (AAV)-Delivered Exosomal TAT and BiTE Molecule CD4-αCD3 Facilitate the Elimination of CD4 T Cells Harboring Latent HIV-1" Microorganisms 12, no. 8: 1707. https://doi.org/10.3390/microorganisms12081707