Orf Virus-Based Vaccine Vector D1701-V Induces Strong CD8+ T Cell Response against the Transgene but Not against ORFV-Derived Epitopes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Viruses
2.2. Infection of Cells
2.3. Flow Cytometry Analysis of Infected Cells
2.4. Isolation of MHC I-Bound Peptides and Mass Spectrometric Analysis
2.5. Database Search and Filtering
2.6. Peptide Synthesis
2.7. Immunization of Mice
2.8. IFN-γ ELISPOT Assay
2.9. MHC Dextramer Staining
2.10. Intracellular Cytokine Staining (ICS)
2.11. In Vitro Re-Stimulation of Splenocytes
2.12. Statistical Analysis
3. Results
3.1. ORFV Vector Strain D1701-V Efficiently Induces Transgene-Specific CD8+ T Cell Response
3.2. HeLa-Kb Cells Are Suitable Targets for the Identification of ORFV-Derived Peptides
3.3. Identification of ORFV-Derived H-2Kb-Presented Peptides by LC-MS/MS
3.4. Characteristics of ORFV-Derived H-2Kb-Presented Peptides
3.5. Characteristics of ORFV-Derived H-2Kb-Presented Peptides
3.6. ICS Validates Absence of CD8+ T Cell Responses to the Individual ORFV-Derived Peptides
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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No. | Sequence | Length (aa) | UniProt Accession No. | Antigen/Protein * | Position | NetMHC Binding Affinity (nM) | NetMHC % Rank of Predicted Affinity |
---|---|---|---|---|---|---|---|
1 | AIYGFGVTF | 9 | ADY76886.1; ADY76885.1 | ORFV056 RNA-polymerase subunit RPO147 | 569–577 | 248.6 | 0.5757 |
2 | VGYPRQNAV | 9 | ADY76846.1; ADY76844.1 | ORFV097 DNA-polymerase processivity factor | 364–372 | 95.7 | 0.2477 |
3 | IGYMVKNL | 8 | ADY76734.1 | ORFV075 Rifampicin resistance protein | 212–219 | 19.1 | 0.0441 |
4 | VGFVHPIAM | 9 | ADY76731.1 | ORF079 Virion core protein P4b | 258–266 | 51.9 | 0.1255 |
5 | SILKFEERL | 9 | ADY76728.1 | ORF083 Early transcription factor VETFL | 286–294 | 93.8 | 0.2427 |
6 | LNLMYPNI | 8 | ADY76725.1 | ORFV86 Virion core precursor protein P4a | 172–179 | 26.7 | 0.0674 |
7 | SGSVPYARL | 9 | ADY76722.1 | ORFV090 IMV membrane protein | 73–81 | 140.6 | 0.3490 |
8 | AAFEFRDL | 8 | ADY76890.1 | ORF052 putative IMV membrane protein | 72–79 | 12.6 | 0.0253 |
9 | VGFVHPIA | 8 | ADY76731.1 | ORF079 Virion core protein P4b | 258–265 | 485.9 | 0.9853 |
10 | KILAPFNFL | 9 | ADY76886.1; ADY76885.1 | ORFV056 RNA-polymerase subunit RPO147 | 834–842 | 385.2 | 0.8315 |
11 | ISALFKQL | 8 | ADY76725.1; ADY76726.1 | ORFV086 Virion core protein P4a | 781–788 | 14.2 | 0.0296 |
12 | TEFPVFEEL | 9 | ADY76883.1 | ORFV058 (IMV, viral entry) | 182–190 | 3095.8 | 4.6680 |
13 | VNIVRQEEL | 9 | ADY76792.1 | ORFV016 Unknown | 35–43 | 270.1 | 0.6179 |
14 | AIIKYTDL | 8 | ADY76837.1 | ORFV110 EEV glycoprotein | 37–44 | 89.3 | 0.2306 |
15 | SILERYNLF | 9 | ADY76725.1; ADY76726.1 | ORFV086 Virion core protein P4a | 810–818 | 130.5 | 0.3285 |
16 | VFFRVTVL | 8 | ADY76779.1 | ORFV028 DNA-binding protein | 644–651 | 73.4 | 0.1860 |
17 | GSVPYARL | 8 | ADY76722.1 | ORFV090 IMV membrane protein | 74–81 | 55.2 | 0.1387 |
18 | QNYSYSERLL | 10 | ADY76809.1 | ORFV129 Ankyrin/F-box protein | 115–124 | 180.9 | 0.4297 |
19 | RVNTFTAV | 8 | ADY76793.1 | ORFV015 Unknown | 33–40 | 738.0 | 1.4130 |
20 | TAVDFTQFL | 9 | ADY76778.1 | ORFV029 Unknown | 244–252 | 172.0 | 0.4073 |
21 | QNYSYSERL | 9 | ADY76809.1 | ORFV129 Ankyrin/F-box protein | 115–123 | 24.3 | 0.0592 |
22 | AIYAFRLT | 8 | ADY76719.1 | ORFV094 Phosphorylated IMV membrane protein | 160–167 | 150.9 | 0.3685 |
23 | ANVDFMEYV | 9 | ADY76886.1; ADY76885.1 | ORFV056 RNA-polymerase subunit RPO147 | 880–888 | 397.0 | 0.8488 |
24 | ISVMFNNV | 8 | ADY76728.1 | ORF083 Early transcription factor VETFL | 484–491 | 10.5 | 0.0198 |
25 | VIFGRQPSL | 9 | ADY76886.1; ADY76885.1 | ORFV056 RNA-polymerase subunit RPO147 | 374–382 | 59.9 | 0.1538 |
26 | EQFSFSNV | 8 | ADY76841.1 | ORFV101 RNA-polymerase subunit RPO132 | 623–630 | 259.0 | 0.5952 |
27 | LIREFANL | 8 | ADY76728.1 | ORF083 Early transcription factor VETFL | 294–301 | 19.5 | 0.0456 |
28 | IAPQLRSL | 8 | ADY76724.1 | ORF088 Virion core protein | 24–31 | 418.0 | 0.8785 |
29 | ISIPRSVGF | 9 | ADY76841.1 | ORFV101 RNA-polymerase subunit RPO132 | 427–435 | 195.8 | 0.4646 |
30 | SIAPMNTGF | 9 | ADY76778.1 | ORFV029 Unknown | 216–224 | 3273.7 | 4.8979 |
31 | IAPQLRSLL | 9 | ADY76724.1 | ORF088 Virion core protein | 24–32 | 331.2 | 0.7371 |
32 | SATQFQSV | 8 | ADY76731.1 | ORF079 Virion core protein P4b | 230–237 | 482.6 | 0.9800 |
33 | SFVVVAQI | 8 | ADY76769.1 | ORFV040 Glutaredoxin-like protein | 98–105 | 2254.5 | 3.5474 |
34 | SIVSFKPTL | 9 | ADY76731.1 | ORF079 Virion core protein P4b | 386–394 | 75.0 | 0.1902 |
35 | TNVEIGKL | 8 | ADY76729.1 | ORFV082 Unknown | 277–284 | 1825.4 | 2.9640 |
36 | VIEIFKQL | 8 | ADY76732.1 | ORFV077 Late transcription factor VLTF-3 | 130–137 | 216.0 | 0.5087 |
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Reguzova, A.; Ghosh, M.; Müller, M.; Rziha, H.-J.; Amann, R. Orf Virus-Based Vaccine Vector D1701-V Induces Strong CD8+ T Cell Response against the Transgene but Not against ORFV-Derived Epitopes. Vaccines 2020, 8, 295. https://doi.org/10.3390/vaccines8020295
Reguzova A, Ghosh M, Müller M, Rziha H-J, Amann R. Orf Virus-Based Vaccine Vector D1701-V Induces Strong CD8+ T Cell Response against the Transgene but Not against ORFV-Derived Epitopes. Vaccines. 2020; 8(2):295. https://doi.org/10.3390/vaccines8020295
Chicago/Turabian StyleReguzova, Alena, Michael Ghosh, Melanie Müller, Hanns-Joachim Rziha, and Ralf Amann. 2020. "Orf Virus-Based Vaccine Vector D1701-V Induces Strong CD8+ T Cell Response against the Transgene but Not against ORFV-Derived Epitopes" Vaccines 8, no. 2: 295. https://doi.org/10.3390/vaccines8020295
APA StyleReguzova, A., Ghosh, M., Müller, M., Rziha, H. -J., & Amann, R. (2020). Orf Virus-Based Vaccine Vector D1701-V Induces Strong CD8+ T Cell Response against the Transgene but Not against ORFV-Derived Epitopes. Vaccines, 8(2), 295. https://doi.org/10.3390/vaccines8020295