Fc-Mediated E2-Dimer Subunit Vaccines of Atypical Porcine Pestivirus Induce Efficient Humoral and Cellular Immune Responses in Piglets
Abstract
:1. Introduction
2. Materials and Methods
2.1. Construction of Recombinant S2 Cell Lines
2.2. Expression and Purification of APPV E2, E2Fc, and E2ΔFc Fusion Proteins
2.3. Co-localization Detection of APPV E2Fc and E2ΔFc Fusion Proteins with FcγRI
2.4. Antigenicity and Affinity Studies of APPV E2, E2Fc, and E2ΔFc Fusion Proteins
2.5. Vaccination of Piglets
2.6. Detection of APPV E2-Specific Antibodies
2.7. Lymphocyte Proliferation Assay
2.8. Detection of Cytokines
2.9. Statistical Analysis
3. Results
3.1. Construction of Recombinant Drosophila Cell Lines
3.2. Purification of APPV E2, E2Fc, and E2ΔFc Fusion Proteins
3.3. Characterization of APPV E2, E2Fc, and E2ΔFc Dimer
3.4. Binding of APPV E2Fc and E2ΔFc Fusion Proteins to FcγRI on APCs
3.5. Antigenicity and Affinity Studies of APPV E2, E2Fc, and E2ΔFc Fusion Proteins
3.6. APPV E2Fc or E2ΔFc Immunization Induce an Efficient Antibody Response in Piglets
3.7. Lymphocyte Proliferative Response
3.8. APPV E2ΔFc Immunization Improves the Th2-Biased Cellular Immune Response
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Acknowledgments
Conflicts of Interest
References
- Kinsley, A.T. Dancing pigs? Vet. Med. 1922, 17, 123. [Google Scholar]
- Arruda, B.L.; Arruda, P.H.; Magstadt, D.R.; Schwartz, K.J.; Dohlman, T.; Schleining, J.A.; Patterson, A.R.; Visek, C.A.; Victoria, J.G. Identification of a Divergent Lineage Porcine Pestivirus in Nursing Piglets with Congenital Tremors and Reproduction of Disease following Experimental Inoculation. PLoS ONE 2016, 11, e0150104. [Google Scholar] [CrossRef] [Green Version]
- De Groof, A.; Deijs, M.; Guelen, L.; van Grinsven, L.; van Os-Galdos, L.; Vogels, W.; Derks, C.; Cruijsen, T.; Geurts, V.; Vrijenhoek, M.; et al. Atypical Porcine Pestivirus: A Possible Cause of Congenital Tremor Type A-II in Newborn Piglets. Viruses 2016, 8, 271. [Google Scholar] [CrossRef]
- Postel, A.; Hansmann, F.; Baechlein, C.; Fischer, N.; Alawi, M.; Grundhoff, A.; Derking, S.; Tenhündfeld, J.; Pfankuche, V.M.; Herder, V.; et al. Presence of atypical porcine pestivirus (APPV) genomes in newborn piglets correlates with congenital tremor. Sci. Rep. 2016, 6, 27735. [Google Scholar] [CrossRef] [Green Version]
- Blomström, A.L.; Ley, C.; Jacobson, M. Astrovirus as a possible cause of congenital tremor type AII in piglets? Acta Vet. Scand. 2014, 56, 82. [Google Scholar] [CrossRef] [Green Version]
- Possatti, F.; Headley, S.A.; Leme, R.A.; Dall Agnol, A.M.; Zotti, E.; de Oliveira, T.E.S.; Alfieri, A.F.; Alfieri, A.A. Viruses associated with congenital tremor and high lethality in piglets. Transbound. Emerg. Dis. 2018, 65, 331–337. [Google Scholar] [CrossRef] [PubMed]
- Schwarz, L.; Riedel, C.; Högler, S.; Sinn, L.J.; Voglmayr, T.; Wöchtl, B.; Dinhopl, N.; Rebel-Bauder, B.; Weissenböck, H.; Ladinig, A.; et al. Congenital infection with atypical porcine pestivirus (APPV) is associated with disease and viral persistence. Vet. Res. 2017, 48, 1. [Google Scholar] [CrossRef] [Green Version]
- Wen, L.; Mao, A.; Jiao, F.; Zhang, D.; Xie, J.; He, K. Evidence of porcine circovirus-like virus P1 in piglets with an unusual congenital tremor. Transbound. Emerg. Dis. 2018, 65, e501–e504. [Google Scholar] [CrossRef] [PubMed]
- Hause, B.M.; Collin, E.A.; Peddireddi, L.; Yuan, F.; Chen, Z.; Hesse, R.A.; Gauger, P.C.; Clement, T.; Fang, Y.; Anderson, G. Discovery of a novel putative atypical porcine pestivirus in pigs in the USA. J. Gen. Virol. 2015, 96, 2994–2998. [Google Scholar] [CrossRef] [PubMed]
- Lamp, B.; Riedel, C.; Wentz, E.; Tortorici, M.A.; Rümenapf, T. Autocatalytic cleavage within classical swine fever virus NS3 leads to a functional separation of protease and helicase. J. Virol. 2013, 87, 11872–11883. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pan, S.; Mou, C.; Chen, Z. An emerging novel virus: Atypical porcine pestivirus (APPV). Rev. Med. Virol. 2019, 29, e2018. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Stark, R.; Meyers, G.; Rümenapf, T.; Thiel, H.J. Processing of pestivirus polyprotein: Cleavage site between autoprotease and nucleocapsid protein of classical swine fever virus. J. Virol. 1993, 67, 7088–7095. [Google Scholar] [CrossRef] [Green Version]
- Tautz, N.; Tews, B.A.; Meyers, G. The Molecular Biology of Pestiviruses. Adv. Virus. Res. 2015, 93, 47–160. [Google Scholar]
- Zhang, H.; Wen, W.; Hao, G.; Chen, H.; Qian, P.; Li, X. A Subunit Vaccine Based on E2 Protein of Atypical Porcine Pestivirus Induces Th2-type Immune Response in Mice. Viruses 2018, 10, 673. [Google Scholar] [CrossRef] [Green Version]
- Liu, J.; Zhang, P.; Chen, Y.; Zhong, W.; Li, B.; Pi, M.; Ning, Z. Vaccination with virus-like particles of atypical porcine pestivirus inhibits virus replication in tissues of BALB/c mice. Arch. Virol. 2021, 166, 2733–2741. [Google Scholar] [CrossRef]
- Li, J.; Li, X.; Ma, H.; Ren, X.; Hao, G.; Zhang, H.; Zhao, Z.; Fang, K.; Li, X.; Rong, Z.; et al. Efficient mucosal vaccination of a novel classical swine fever virus E2-Fc fusion protein mediated by neonatal Fc receptor. Vaccine 2020, 38, 4574–4583. [Google Scholar] [CrossRef] [PubMed]
- Yuan, J.; Han, Z.; Li, J.; Huang, Y.; Yang, J.; Ding, H.; Zhang, J.; Zhu, M.; Zhang, Y.; Liao, J.; et al. Atypical Porcine Pestivirus as a Novel Type of Pestivirus in Pigs in China. Front. Microbiol. 2017, 8, 862. [Google Scholar] [CrossRef] [PubMed]
- Pan, S.; Yan, Y.; Shi, K.; Wang, M.; Mou, C.; Chen, Z. Molecular characterization of two novel atypical porcine pestivirus (APPV) strains from piglets with congenital tremor in China. Transbound. Emerg. Dis. 2019, 66, 35–42. [Google Scholar] [CrossRef]
- Yan, X.L.; Li, Y.Y.; He, L.L.; Wu, J.L.; Tang, X.Y.; Chen, G.H.; Mai, K.J.; Wu, R.T.; Li, Q.N.; Chen, Y.H.; et al. 12 novel atypical porcine pestivirus genomes from neonatal piglets with congenital tremors: A newly emerging branch and high prevalence in China. Virology 2019, 533, 50–58. [Google Scholar] [CrossRef]
- Xie, Y.; Wang, X.; Su, D.; Feng, J.; Wei, L.; Cai, W.; Li, J.; Lin, S.; Yan, H.; He, D. Detection and Genetic Characterization of Atypical Porcine Pestivirus in Piglets with Congenital Tremors in Southern China. Front. Microbiol. 2019, 10, 1406. [Google Scholar] [CrossRef] [Green Version]
- Mackness, B.C.; Jaworski, J.A.; Boudanova, E.; Park, A.; Valente, D.; Mauriac, C.; Pasquier, O.; Schmidt, T.; Kabiri, M.; Kandira, A.; et al. Antibody Fc engineering for enhanced neonatal Fc receptor binding and prolonged circulation half-life. MAbs 2019, 11, 1276–1288. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ying, T.; Chen, W.; Feng, Y.; Wang, Y.; Gong, R.; Dimitrov, D.S. Engineered soluble monomeric IgG1 CH3 domain: Generation, mechanisms of function, and implications for design of biological therapeutics. J. Biol. Chem. 2013, 288, 25154–25164. [Google Scholar] [CrossRef] [Green Version]
- Ying, T.; Chen, W.; Gong, R.; Feng, Y.; Dimitrov, D.S. Soluble monomeric IgG1 Fc. J. Biol. Chem. 2012, 287, 19399–19408. [Google Scholar] [CrossRef] [Green Version]
- Wang, Y.; Tian, Z.; Thirumalai, D.; Zhang, X. Neonatal Fc receptor (FcRn): A novel target for therapeutic antibodies and antibody engineering. J. Drug. Target. 2014, 22, 269–278. [Google Scholar] [CrossRef]
- Liu, J.; Li, Z.; Ren, X.; Li, H.; Lu, R.; Zhang, Y.; Ning, Z. Viral load and histological distribution of atypical porcine pestivirus in different tissues of naturally infected piglets. Arch. Virol. 2019, 164, 2519–2523. [Google Scholar] [CrossRef] [PubMed]
- Cagatay, G.N.; Meyer, D.; Wendt, M.; Becher, P.; Postel, A. Characterization of the Humoral Immune Response Induced after Infection with Atypical Porcine Pestivirus (APPV). Viruses 2019, 11, 880. [Google Scholar] [CrossRef] [PubMed]
- Zhang, K.; Wu, K.; Liu, J.; Ge, S.; Xiao, Y.; Shang, Y.; Ning, Z. Identification of atypical porcine pestivirus infection in swine herds in China. Transbound. Emerg. Dis. 2017, 64, 1020–1023. [Google Scholar] [CrossRef]
- Zhang, H.; Wen, W.; Hao, G.; Hu, Y.; Chen, H.; Qian, P.; Li, X. Phylogenetic and genomic characterization of a novel atypical porcine pestivirus in China. Transbound. Emerg. Dis. 2018, 65, e202–e204. [Google Scholar] [CrossRef] [PubMed]
- Wang, Z.; Nie, Y.; Wang, P.; Ding, M.; Deng, H. Characterization of classical swine fever virus entry by using pseudotyped viruses: E1 and E2 are sufficient to mediate viral entry. Virology 2004, 330, 332–341. [Google Scholar] [CrossRef] [Green Version]
- Reimann, I.; Depner, K.; Trapp, S.; Beer, M. An avirulent chimeric Pestivirus with altered cell tropism protects pigs against lethal infection with classical swine fever virus. Virology 2004, 322, 143–157. [Google Scholar] [CrossRef] [Green Version]
- Risatti, G.R.; Borca, M.V.; Kutish, G.F.; Lu, Z.; Holinka, L.G.; French, R.A.; Tulman, E.R.; Rock, D.L. The E2 glycoprotein of classical swine fever virus is a virulence determinant in swine. J. Virol. 2005, 79, 3787–3796. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Suzuki, T.; Ishii-Watabe, A.; Tada, M.; Kobayashi, T.; Kanayasu-Toyoda, T.; Kawanishi, T.; Yamaguchi, T. Importance of neonatal FcR in regulating the serum half-life of therapeutic proteins containing the Fc domain of human IgG1: A comparative study of the affinity of monoclonal antibodies and Fc-fusion proteins to human neonatal FcR. J. Immunol. 2010, 184, 1968–1976. [Google Scholar] [CrossRef] [PubMed]
- Abdiche, Y.N.; Yeung, Y.A.; Chaparro-Riggers, J.; Barman, I.; Strop, P.; Chin, S.M.; Pham, A.; Bolton, G.; McDonough, D.; Lindquist, K.; et al. The neonatal Fc receptor (FcRn) binds independently to both sites of the IgG homodimer with identical affinity. MAbs 2015, 7, 331–343. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Czajkowsky, D.M.; Hu, J.; Shao, Z.; Pleass, R.J. Fc-fusion proteins: New developments and future perspectives. EMBO Mol. Med. 2012, 4, 1015–1028. [Google Scholar] [CrossRef]
- Nie, J.; Li, Q.; Wu, J.; Zhao, C.; Hao, H.; Liu, H.; Zhang, L.; Nie, L.; Qin, H.; Wang, M.; et al. Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2. Emerg. Microbes. Infect. 2020, 9, 680–686. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xiong, H.L.; Wu, Y.T.; Cao, J.L.; Yang, R.; Liu, Y.X.; Ma, J.; Qiao, X.Y.; Yao, X.Y.; Zhang, B.H.; Zhang, Y.L.; et al. Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells. Emerg. Microbes. Infect. 2020, 9, 2105–2113. [Google Scholar] [CrossRef] [PubMed]
- Schmidt, F.; Weisblum, Y.; Muecksch, F.; Hoffmann, H.H.; Michailidis, E.; Lorenzi, J.C.C.; Mendoza, P.; Rutkowska, M.; Bednarski, E.; Gaebler, C.; et al. Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses. J. Exp. Med. 2020, 217, e20201181. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Q.; Yu, Y.; Song, C.; Cao, Z. Genetic characterization of atypical porcine pestivirus (APPV) in China and the successful isolation of a novel APPV strain within genotype 2. Microb. Pathog. 2021, 161 Pt A, 105282. [Google Scholar] [CrossRef]
Experimental Group | Number of Pigs | Antigen | Dose (μg/Pig) | Adjuvant | Delivery Route |
---|---|---|---|---|---|
A | 5 | aE2 | 80 | ISA 201VG | Intramuscular |
B | 5 | aE2 | 80 | IMS 1313VG | Intramuscular |
C | 5 | aE2Fc | 80 | ISA 201VG | Intramuscular |
D | 5 | aE2Fc | 80 | IMS 1313VG | Intramuscular |
E | 5 | aE2ΔFc | 80 | ISA 201VG | Intramuscular |
F | 5 | aE2ΔFc | 80 | IMS 1313VG | Intramuscular |
G | 3 | PBS | 2 mL | - | Intramuscular |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Ren, X.; Qian, P.; Liu, S.; Chen, H.; Li, X. Fc-Mediated E2-Dimer Subunit Vaccines of Atypical Porcine Pestivirus Induce Efficient Humoral and Cellular Immune Responses in Piglets. Viruses 2021, 13, 2443. https://doi.org/10.3390/v13122443
Ren X, Qian P, Liu S, Chen H, Li X. Fc-Mediated E2-Dimer Subunit Vaccines of Atypical Porcine Pestivirus Induce Efficient Humoral and Cellular Immune Responses in Piglets. Viruses. 2021; 13(12):2443. https://doi.org/10.3390/v13122443
Chicago/Turabian StyleRen, Xujiao, Ping Qian, Shudan Liu, Huanchun Chen, and Xiangmin Li. 2021. "Fc-Mediated E2-Dimer Subunit Vaccines of Atypical Porcine Pestivirus Induce Efficient Humoral and Cellular Immune Responses in Piglets" Viruses 13, no. 12: 2443. https://doi.org/10.3390/v13122443
APA StyleRen, X., Qian, P., Liu, S., Chen, H., & Li, X. (2021). Fc-Mediated E2-Dimer Subunit Vaccines of Atypical Porcine Pestivirus Induce Efficient Humoral and Cellular Immune Responses in Piglets. Viruses, 13(12), 2443. https://doi.org/10.3390/v13122443