Lysosome-Associated Membrane Protein Targeting Strategy Improved Immunogenicity of Glycoprotein-Based DNA Vaccine for Marburg Virus
Abstract
:1. Introduction
2. Materials and Methods
2.1. Recombinant Plasmid
2.2. Transfection of HEK293T Cells
2.3. Quantitative Real-Time PCR
2.4. Western Blot
2.5. Immunofluorescence Analysis
2.6. Prokaryotic Expression of MARV GP
2.7. Animals and Immunization
2.8. Enzyme-Linked Immunosorbent Assay (ELISA)
2.9. Serum Neutralization Test
2.10. Synthesis of GP Peptides
2.11. Enzyme-Linked Immunospot Assay (ELISpot)
2.12. Flow Cytometry
2.13. Transcriptome Analysis
2.14. Animal Behavior Analysis
2.15. Hematoxylin and Eosin (H&E) Staining
2.16. Statistical Analysis
3. Results
3.1. Construction and Verification of Plasmids
3.2. pVAX1-LAMP/GPMARV Induced a Stronger Humoral Immune Response
3.3. The LAMP Targeting Strategy Enhanced the Secretion of IFN-γ and IL-4
3.4. Flow Cytometry Revealed That LAMP1 Can Enhance the T-Cell Response
3.5. Transcriptome Analysis of Immune Response-Related Pathways
3.6. Preliminary Safety Assessment of Candidate Vaccines
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Zhang, X.; Sun, Y.; Zhang, J.; Wei, H.; Wang, J.; Hu, C.; Liu, Y.; Cai, S.; Yuan, Q.; Wang, Y.; et al. Lysosome-Associated Membrane Protein Targeting Strategy Improved Immunogenicity of Glycoprotein-Based DNA Vaccine for Marburg Virus. Vaccines 2024, 12, 1013. https://doi.org/10.3390/vaccines12091013
Zhang X, Sun Y, Zhang J, Wei H, Wang J, Hu C, Liu Y, Cai S, Yuan Q, Wang Y, et al. Lysosome-Associated Membrane Protein Targeting Strategy Improved Immunogenicity of Glycoprotein-Based DNA Vaccine for Marburg Virus. Vaccines. 2024; 12(9):1013. https://doi.org/10.3390/vaccines12091013
Chicago/Turabian StyleZhang, Xiyang, Yubo Sun, Junqi Zhang, Hengzheng Wei, Jing Wang, Chenchen Hu, Yang Liu, Sirui Cai, Qinghong Yuan, Yueyue Wang, and et al. 2024. "Lysosome-Associated Membrane Protein Targeting Strategy Improved Immunogenicity of Glycoprotein-Based DNA Vaccine for Marburg Virus" Vaccines 12, no. 9: 1013. https://doi.org/10.3390/vaccines12091013