An Equine Model for Vaccination against a Hepacivirus: Insights into Host Responses to E2 Recombinant Protein Vaccination and Subsequent Equine Hepacivirus Inoculation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Animals
2.2. EqHV E2 Recombinant Protein Production
2.3. Vaccination and EqHV Inoculation
2.4. Sample Collection
2.5. Laboratory Analysis
2.5.1. Detection of EqHV RNA in Serum
2.5.2. Detection and Isotyping of Anti-EqHV E2-specific IgG (E2 IgG) in Serum
2.5.3. Detection of Anti-EqHV NS3-Specific IgG in Serum
2.5.4. Monitoring of Haematology, Serum Biochemistry, and Clinical Parameters
2.5.5. 1H Nuclear Magnetic Resonance (NMR) Spectroscopy of Serum Samples
2.5.6. RNA Extraction and RNA Sequencing (RNA-seq) of Liver Tissue
2.5.7. Histopathology, Immunohistochemistry (IHC), and Fluorescent in Situ Hybridization (FISH) of Liver Tissue
2.5.8. Detection of EqHV RNA in Liver Tissue
3. Results
3.1. EqHV RNA in Serum of Vaccine Ponies and Control Ponies
3.2. Anti-EqHV E2-Specific IgG Isotypes in Serum of Vaccine Ponies and Control Ponies
3.3. Anti-EqHV NS3-Specific IgG in Serum of Vaccine Ponies and Control Ponies
3.4. Liver-Associated Serum Biochemistry Parameters
3.5. Clinical Scoring
3.6. Serum Metabolomics
3.7. RNA-seq of Liver Tissue
3.8. Histopathology, IHC, FISH, and qPCR of Liver Tissue
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Blood Parameters Score Overview | ||||||
---|---|---|---|---|---|---|
Vaccine Pony 1 | Vaccine Pony 2 | Vaccine Pony 3 | Vaccine Pony 4 | Control Pony 1 | Control Pony 2 | |
D+7 | 0 | 0 | 0 | 1 | 1 | 0 |
D+14 | 1 | 0 | 0 | 1 | 2 | 4 |
D+21 | 0 | 0 | 0 | 1 | 0 | 0 |
D+28 | 0 | 4 | 0 | 0 | 1 | 0 |
D+35 | 0 | 0 | 1 | 0 | 1 | 0 |
D+42 | 0 | 0 | 3 | 1 | 1 | 0 |
D+49 | 1 | 2 | 5 | 1 | 3 | 0 |
D+56 | 4 | 1 | 5 | 0 | 0 | 1 |
D+63 | 5 | 5 | 4 | 1 | 1 | 3 |
D+70 | 3 | 4 | 3 | 1 | 2 | 4 |
D+78 | 1 | 4 | 2 | 2 | 3 | 5 |
D+84 | 1 | 3 | 1 | 3 | 5 | 5 |
D+90 | 1 | 2 | 2 | 4 | 6 | 3 |
D+98 | 2 | 1 | 1 | 4 | 5 | 3 |
D+105 | 1 | 1 | 1 | 2 | 5 | 3 |
D+112 | 0 | 0 | 1 | 1 | 5 | 2 |
Total blood parameters score | 20 | 27 | 29 | 23 | 41 | 33 |
+ Total clinical parameters score | 0 | 0 | 2 | 0 | 0 | 0 |
Total overall clinical score | 20 | 27 | 31 | 23 | 41 | 33 |
Metabolite | Mean Difference | t | p Value | Adj. p Value |
---|---|---|---|---|
Pyruvate | 0.027 | 5.334 | 0.000 | 0.001 |
Acetoacetate | −0.007 | −5.258 | 0.000 | 0.001 |
Formate | −0.013 | −5.177 | 0.000 | 0.001 |
Methanol | −0.058 | −5.153 | 0.000 | 0.001 |
Albumin | −0.168 | −3.647 | 0.001 | 0.017 |
Tot.CCE | −12.367 | −3.532 | 0.002 | 0.018 |
Total protein | −0.064 | −3.458 | 0.002 | 0.018 |
Carbon chains | −2.594 | −3.443 | 0.002 | 0.018 |
Sphingomyelins | −0.175 | −3.283 | 0.003 | 0.023 |
Sub8.Tot.Chol. | −0.024 | −3.087 | 0.005 | 0.031 |
Fatty acids | −0.514 | −3.072 | 0.006 | 0.031 |
Sub7.Tot.Chol. | −0.023 | −3.039 | 0.006 | 0.031 |
Triglycerids | −0.203 | −2.946 | 0.008 | 0.032 |
Multi unsat.CCE | −0.652 | −2.899 | 0.008 | 0.032 |
Histidine | −0.013 | −2.871 | 0.009 | 0.032 |
Alpha unsat.CCE | −1.477 | −2.842 | 0.010 | 0.032 |
Glutamine | −0.091 | −2.829 | 0.010 | 0.032 |
Unsat.CCE | −2.496 | −2.820 | 0.010 | 0.032 |
Sub9.Tot.Chol. | −0.018 | −2.817 | 0.010 | 0.032 |
Tot.cholesterol | −0.127 | −2.737 | 0.012 | 0.035 |
Creatinine | −0.037 | −2.719 | 0.013 | 0.035 |
Mannose | 0.014 | 2.711 | 0.013 | 0.035 |
Lactate | 0.425 | 2.579 | 0.017 | 0.045 |
Sub6.Tot.Chol. | −0.015 | −2.537 | 0.019 | 0.047 |
Mean Liver Viral Load (Viral Copies/50 ng Total RNA; LLOQ = 1 × 103 Copies/50 ng Total RNA) | Serum Viral Load (Viral Copies/mL Serum; LLOQ = 7.2 × 104 Copies/mL Serum) | |
---|---|---|
Day +13 | Day +14 | |
Vaccine Pony 1 | 8.72 × 104 | 9.51 × 106 |
Vaccine Pony 2 | 3.55 × 104 | 1.14 × 107 |
Vaccine Pony 3 | 7.52 × 104 | 1.36 × 107 |
Vaccine Pony 4 | 1.30 × 105 | 1.47 × 107 |
Control Pony 1 | 8.77 × 104 | 1.50 × 107 |
Control Pony 2 | 7.66 × 104 | 9.80 × 106 |
Day +97 | Day +98 | |
Vaccine Pony 1 | 1.27 × 103 | <LLOQ |
Vaccine Pony 2 | <LLOQ | <LLOQ |
Vaccine Pony 3 | <LLOQ c | <LLOQ |
Vaccine Pony 4 | 1.00 × 104 | 4.66 × 105 |
Control Pony 1 | 9.28 × 103 | 4.13 × 106 |
Control Pony 2 | <LLOQ | 3.01 × 105 |
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Badenhorst, M.; Saalmüller, A.; Daly, J.M.; Ertl, R.; Stadler, M.; Puff, C.; de le Roi, M.; Baumgärtner, W.; Engelmann, M.; Brandner, S.; et al. An Equine Model for Vaccination against a Hepacivirus: Insights into Host Responses to E2 Recombinant Protein Vaccination and Subsequent Equine Hepacivirus Inoculation. Viruses 2022, 14, 1401. https://doi.org/10.3390/v14071401
Badenhorst M, Saalmüller A, Daly JM, Ertl R, Stadler M, Puff C, de le Roi M, Baumgärtner W, Engelmann M, Brandner S, et al. An Equine Model for Vaccination against a Hepacivirus: Insights into Host Responses to E2 Recombinant Protein Vaccination and Subsequent Equine Hepacivirus Inoculation. Viruses. 2022; 14(7):1401. https://doi.org/10.3390/v14071401
Chicago/Turabian StyleBadenhorst, Marcha, Armin Saalmüller, Janet M. Daly, Reinhard Ertl, Maria Stadler, Christina Puff, Madeleine de le Roi, Wolfgang Baumgärtner, Michael Engelmann, Sabine Brandner, and et al. 2022. "An Equine Model for Vaccination against a Hepacivirus: Insights into Host Responses to E2 Recombinant Protein Vaccination and Subsequent Equine Hepacivirus Inoculation" Viruses 14, no. 7: 1401. https://doi.org/10.3390/v14071401