Nanoparticles Carrying Conserved Regions of Influenza A Hemagglutinin, Nucleoprotein, and M2 Protein Elicit a Strong Humoral and T Cell Immune Response and Protect Animals from Infection
Abstract
:1. Introduction
2. Results
2.1. Design of Recombinant Proteins
2.2. Expression and Purification of Recombinant Proteins
2.3. Formation of Nanoparticles
2.4. Humoral Immune Response
2.5. T Cellular Immune Response
2.6. Protection against Lethal Influenza Challenge
3. Discussion
4. Materials and Methods
4.1. Components of SAP-Based Fusion Proteins
4.2. Genes Encoding Fusion Proteins and Expression Vectors
4.3. Expression and Purification of Fusion Proteins
4.4. Western Blot Analysis of Protein Samples
4.5. Structural Analysis of Recombinant Proteins
4.6. Immunization of Animals
4.7. Antibody Detection in the Sera and BAL by ELISA
4.8. Isolation of Cells from the Spleen to Analyze the T Cellular Immune Response
4.9. Determination of Antigen-Specific Effector Memory T cells
4.10. Influenza Virus and Challenge Experiment
4.11. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Group of Mice | M2e-Specific Tem Cells | Virus-Specific Tem Cells | ||
---|---|---|---|---|
CD4+ | CD8+ | CD4+ | CD8+ | |
Experimental | 0.28 ± 0.07% | 0.276 ± 0.11% | 5.98 ± 0.63% | 13.69 ± 1.74% |
Control (PBS) | 0.04 ± 0.02% | 0.08 ± 0.05% | 4.37 ± 0.63% | 8.60 ± 1.00% |
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Zykova, A.A.; Blokhina, E.A.; Stepanova, L.A.; Shuklina, M.A.; Ozhereleva, O.O.; Tsybalova, L.M.; Kuprianov, V.V.; Ravin, N.V. Nanoparticles Carrying Conserved Regions of Influenza A Hemagglutinin, Nucleoprotein, and M2 Protein Elicit a Strong Humoral and T Cell Immune Response and Protect Animals from Infection. Molecules 2023, 28, 6441. https://doi.org/10.3390/molecules28186441
Zykova AA, Blokhina EA, Stepanova LA, Shuklina MA, Ozhereleva OO, Tsybalova LM, Kuprianov VV, Ravin NV. Nanoparticles Carrying Conserved Regions of Influenza A Hemagglutinin, Nucleoprotein, and M2 Protein Elicit a Strong Humoral and T Cell Immune Response and Protect Animals from Infection. Molecules. 2023; 28(18):6441. https://doi.org/10.3390/molecules28186441
Chicago/Turabian StyleZykova, Anna A., Elena A. Blokhina, Liudmila A. Stepanova, Marina A. Shuklina, Olga O. Ozhereleva, Liudmila M. Tsybalova, Victor V. Kuprianov, and Nikolai V. Ravin. 2023. "Nanoparticles Carrying Conserved Regions of Influenza A Hemagglutinin, Nucleoprotein, and M2 Protein Elicit a Strong Humoral and T Cell Immune Response and Protect Animals from Infection" Molecules 28, no. 18: 6441. https://doi.org/10.3390/molecules28186441