Replication of a Dog-Origin H6N1 Influenza Virus in Cell Culture and Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Viruses
2.2. Cells
2.3. Plaque Assay
2.4. Viral Dynamics
2.5. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) Assay
2.6. Chloramphenicol Acetyl Transferase Enzyme-Linked Immunosorbent Assay (CAT ELISA)
2.7. Animals
2.8. Quantitative Reverse Transcription Real Time PCR (qRT-PCR)
2.9. Histopathological Analysis and Immunohistochemistry (IHC)
2.10. Histopathological and Immunohistochemistry Scoring
2.11. Statistical Analyses
3. Results
3.1. Viral Dynamics and Plaque Assay
3.2. Cellular Viability Comparison
3.3. Viral Ribonucleoprotein (RNP) Activity Test
3.4. The Virus Infection of Mice
3.5. Virus Amount Quantification
3.6. Histopathological and Immunohistochemistry (IHC) Evaluation
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Viruses | Amino Acid | |
---|---|---|
PB1 Position 739 | PB2 Position 627 | |
E01 | E | K |
E01EE | E | E |
E01GK | G | K |
PBS | 104 TCID50/mL of E01EE | 104 TCID50/mL of E01GK | 106 TCID50/mL of E01GK | |
---|---|---|---|---|
H&E b | 0 | 2 | 3 | 3 |
IHC c | 0 | 1 | 2 | 3 |
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Tsai, S.-K.; Shih, C.-H.; Chang, H.-W.; Teng, K.-H.; Hsu, W.-E.; Lin, H.-J.; Lin, H.-Y.; Huang, C.-H.; Chen, H.-W.; Wang, L.-C. Replication of a Dog-Origin H6N1 Influenza Virus in Cell Culture and Mice. Viruses 2020, 12, 704. https://doi.org/10.3390/v12070704
Tsai S-K, Shih C-H, Chang H-W, Teng K-H, Hsu W-E, Lin H-J, Lin H-Y, Huang C-H, Chen H-W, Wang L-C. Replication of a Dog-Origin H6N1 Influenza Virus in Cell Culture and Mice. Viruses. 2020; 12(7):704. https://doi.org/10.3390/v12070704
Chicago/Turabian StyleTsai, Shou-Kuan, Cheng-Hsin Shih, Hui-Wen Chang, Kuang-Huan Teng, Wei-En Hsu, Han-Jia Lin, Han-You Lin, Ching-Huei Huang, Hui-Wen Chen, and Lih-Chiann Wang. 2020. "Replication of a Dog-Origin H6N1 Influenza Virus in Cell Culture and Mice" Viruses 12, no. 7: 704. https://doi.org/10.3390/v12070704