Novel Neutralizing Epitope of PEDV S1 Protein Identified by IgM Monoclonal Antibody
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethics Statement of Animal Use
2.2. Cells and Virus Propagation
2.3. Production of Recombinant S1 Segment of PEDV Spike (S) Protein
2.4. Preparation of Monoclonal Antibodies (mAbs) to the Recombinant S1 (rS1)
2.5. Enzyme-Linked Immunosorbent Assay (ELISA)
2.6. Western Blot Analysis
2.7. Co-localization of the mAbs with S1 Overexpressed in Transfected-Mammalian Cells
2.8. Combined Co-Immunoprecipitation and Western Blotting
2.9. PEDV Neutralization Assay
2.10. Epitope Mapping of the Neutralizing Monoclonal Antibody
2.11. Multiple Sequence Alignment
2.12. Statistical Analysis
3. Results
3.1. Recombinant S1 of PEDV Spike Protein (rS1)
3.2. Hybridoma Generation and Production of mAbs to Recombinant S1 Protein
3.3. Colocalization of mAbs to PEDV S1 Overexpressed in Transfected-HeLa Cells
3.4. Binding of the mAb to Native S Protein
3.5. Neutralization of PEDV Infectivity by the S1 mAb
3.6. Epitope Mapping by Means of Phage Mimotope Identification
3.7. Peptide-Binding ELISA for mAbA3 Epitope Validation
3.8. The Novel A3-Epitope Is Shared among Alphacoronaviruses
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Stevenson, G.W.; Hoang, H.; Schwartz, K.J.; Burrough, E.R.; Sun, D.; Madson, D.; Cooper, V.L.; Pillatzki, A.; Gauger, P.; Schmitt, B.J.; et al. Emergence of porcine epidemic diarrhea virus in the United States: Clinical signs, lesions, and viral genomic sequences. J. Vet. Diagn. Invest. 2013, 25, 649–654. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wood, E.N. An apparently new syndrome of porcine epidemic diarrhoea. Vet. Rec. 1977, 100, 243–244. [Google Scholar] [CrossRef] [PubMed]
- Pensaert, M.B.; De Bouck, P. A new coronavirus-like particle associated with diarrhea in swine. Arch. Virol. 1978, 58, 243–247. [Google Scholar] [CrossRef] [Green Version]
- Lee, C. Porcine epidemic diarrhea virus: An emerging and re-emerging epizootic swine virus. Virol. J. 2015, 12, 193. [Google Scholar] [CrossRef] [Green Version]
- Schulz, L.L.; Tonsor, G.T. Assessment of the economic impacts of porcine epidemic diarrhea virus in the United States. J. Anim. Sci. 2015, 93, 5111–5118. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jung, K.; Saif, L.J.; Wang, Q. Porcine epidemic diarrhea virus (PEDV): An update on etiology, transmission, pathogenesis, and prevention and control. Virus Res. 2020, 286, 198045. [Google Scholar] [CrossRef] [PubMed]
- Song, D.; Park, B. Porcine epidemic diarrhoea virus: A comprehensive review of molecular epidemiology, diagnosis, and vaccines. Virus Genes. 2012, 44, 167–175. [Google Scholar] [CrossRef]
- Gerber, P.F.; Xiao, C.-T.; Lager, K.; Crawford, K.; Kulshreshtha, V.; Cao, D.; Meng, X.; Opriessnig, T. Increased frequency of porcine epidemic diarrhea virus shedding and lesions in suckling pigs compared to nursery pigs and protective immunity in nursery pigs after homologous re-challenge. Vet. Res. 2016, 47, 118. [Google Scholar] [CrossRef] [Green Version]
- Curry, S.M.; Gibson, K.A.; Burrough, E.R.; Schwartz, K.J.; Yoon, K.J.; Gabler, N.K. Nursery pig growth performance and tissue accretion modulation due to porcine epidemic diarrhea virus or porcine deltacoronavirus challenge. J. Anim. Sci. 2017, 95, 173–181. [Google Scholar] [CrossRef] [Green Version]
- Alvarez, J.; Sarradell, J.; Morrison, R.; Perez, A. Impact of porcine epidemic diarrhea on performance of growing pigs. PLoS ONE 2015, 10, e0120532. [Google Scholar] [CrossRef]
- Olanratmanee, E.-O.; Kunavongkrit, A.; Tummaruk, P. Impact of porcine epidemic diarrhea virus infection at different periods of pregnancy on subsequent reproductive performance in gilts and sows. Anim. Reprod. Sci. 2010, 122, 42–51. [Google Scholar] [CrossRef] [PubMed]
- Liu, C.; Tang, J.; Ma, Y.; Liang, X.; Yang, Y.; Peng, G.; Qi, Q.; Jiang, S.; Li, J.; Du, L.; et al. Receptor usage and cell entry of porcine epidemic diarrhea coronavirus. J. Virol. 2015, 89, 6121–6125. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kirchdoerfer, R.N.; Bhandari, M.; Martini, O.; Sewall, L.M.; Bangaru, S.; Yoon, K.-J.; Ward, A.B. Structure and immune recognition of the porcine epidemic diarrhea virus spike protein. Structure 2021, 29, 385–392.e5. [Google Scholar] [CrossRef] [PubMed]
- Li, C.; Li, W.; Lucio de Esesarte, E.; Guo, H.; van den Elzen, P.; Aarts, E.; van den Born, E.; Rottier, P.J.M.; Bosch, B.J. Cell attachment domains of the porcine epidemic diarrhea virus spike protein are key targets of neutralizing antibodies. J. Virol. 2017, 91, e00273-17. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Masuda, T.; Murakami, S.; Takahashi, O.; Miyazaki, A.; Ohashi, S.; Yamasato, H.; Suzuki, T. New porcine epidemic diarrhoea virus variant with a large deletion in the spike gene identified in domestic pigs. Arch. Virol. 2015, 160, 2565–2568. [Google Scholar] [CrossRef] [PubMed]
- Diep, N.V.; Norimine, J.; Sueyoshi, M.; Lan, N.T.; Yamaguchi, R. Novel porcine epidemic diarrhea virus (PEDV) variants with large deletions in the spike (s) gene coexist with PEDV strains possessing an intact s gene in domestic pigs in Japan: A new disease situation. PLoS ONE 2017, 12, e0170126. [Google Scholar] [CrossRef]
- Hou, Y.; Lin, C.-M.; Yokoyama, M.; Yount, B.L.; Marthaler, D.; Douglas, A.L.; Ghimire, S.; Qin, Y.; Baric, R.S.; Saif, L.J.; et al. Deletion of a 197-amino-acid region in the N-terminal domain of spike protein attenuates porcine epidemic diarrhea virus in piglets. J. Virol. 2017, 91, e00227-17. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chang, S.H.; Bae, J.L.; Kang, T.J.; Kim, J.; Chung, G.H.; Lim, C.W.; Laude, H.; Yang, M.S.; Jang, Y.S. Identification of the epitope region capable of inducing neutralizing antibodies against the porcine epidemic diarrhea virus. Mol. Cells. 2002, 14, 295–299. [Google Scholar] [PubMed]
- Sun, D.B.; Feng, L.; Shi, H.Y.; Chen, J.F.; Liu, S.W.; Chen, H.Y.; Wang, Y.F. Spike protein region (aa 636–789) of porcine epidemic diarrhea virus is essential for induction of neutralizing antibodies. Acta Virol. 2007, 51, 149–156. [Google Scholar]
- Okda, F.A.; Lawson, S.; Singrey, A.; Nelson, J.; Hain, K.S.; Joshi, L.R.; Christopher-Hennings, J.; Nelson, E.A.; Diel, D.G. The S2 glycoprotein subunit of porcine epidemic diarrhea virus contains immunodominant neutralizing epitopes. Virology 2017, 509, 185–194. [Google Scholar] [CrossRef]
- Gerdts, V.; Zakhartchouk, A. Vaccines for porcine epidemic diarrhea virus and other swine coronaviruses. Vet. Microbiol. 2017, 206, 45–51. [Google Scholar] [CrossRef]
- Baek, P.-S.; Choi, H.-W.; Lee, S.; Yoon, I.-J.; Lee, Y.J.; Du Lee, S.; Lee, S.; Lee, C. Efficacy of an inactivated genotype 2b porcine epidemic diarrhea virus vaccine in neonatal piglets. Vet. Immunol. Immunopathol. 2016, 174, 45–49. [Google Scholar] [CrossRef]
- Crawford, K.; Mogler, M.; Hicks, J.; Harris, D.L. Protective efficacy of a replicon particle vaccine in both naïve and previously exposed gilts against porcine epidemic diarrhea virus. Ann. Proc. Am. Assoc. Swine Veterinarians. 2016, 212–213. [Google Scholar]
- Greiner, L.; Connor, J.; Graham, A.; Mellor, J.; Lowe, J. Evaluation of a PED vaccine on piglet mortality and sow immunity. In Proceedings of the 46th 2015 Meeting of the American Association of Swine Veterinarians, Orlando, FL, USA, 28 February–3 March 2015; p. 361. [Google Scholar]
- Liu, Y.; Soh, W.T.; Kishikawa, J.I.; Hirose, M.; Nakayama, E.E.; Li, S.; Sasai, M.; Suzuki, T.; Tada, A.; Arakawa, A.; et al. An infectivity-enhancing site on the SARS-CoV-2 spike protein targeted by antibodies. Cell 2021, 184, 3452–3466.e18. [Google Scholar] [CrossRef] [PubMed]
- Zhou, Y.; Liu, Z.; Li, S.; Xu, W.; Zhang, Q.; Silva, I.T.; Li, C.; Wu, Y.; Jiang, Q.; Liu, Z.; et al. Enhancement versus neutralization by SARS-COV-2 antibodies from a convalescent donor associated with distinct epitopes on the RBD. Cell Rep. 2021, 34, 108699. [Google Scholar] [CrossRef] [PubMed]
- Sun, D.; Feng, L.; Shi, H.; Chen, J.; Cui, X.; Chen, H.; Liu, S.; Tong, Y.; Wang, Y.; Tong, G. Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein. Vet. Microbiol. 2008, 131, 73–81. [Google Scholar] [CrossRef] [PubMed]
- Gong, L.; Lin, Y.; Qin, J.; Li, Q.; Xue, C.; Cao, Y. Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization. Virol. J. 2018, 15, 133. [Google Scholar] [CrossRef] [PubMed]
- Chang, C.Y.; Cheng, I.C.; Chang, Y.C.; Tsai, P.S.; Lai, S.Y.; Huang, Y.L.; Jeng, C.R.; Pang, V.F.; Chang, H.W. Identification of neutralizing monoclonal antibodies targeting novel conformational epitopes of the porcine epidemic diarrhoea virus spike protein. Sci. Rep. 2019, 9, 2529. [Google Scholar] [CrossRef] [PubMed]
- Sun, Y.-G.; Li, R.; Xie, S.; Qiao, S.; Li, Q.; Chen, X.X.; Deng, R.; Zhang, G. Identification of a novel linear B-cell epitope within the collagenase equivalent domain of porcine epidemic diarrhea virus spike glycoprotein. Virus Res. 2019, 266, 34–42. [Google Scholar] [CrossRef]
- Kong, N.; Meng, Q.; Jiao, Y.; Wu, Y.; Zuo, Y.; Wang, H.; Sun, D.; Dong, S.; Zhai, H.; Tong, W.; et al. Identification of a novel B-cell epitope in the spike protein of porcine epidemic diarrhea virus. Virol. J. 2020, 17, 46. [Google Scholar] [CrossRef] [Green Version]
- Baer, A.; Kehn-Hall, K. Viral concentration determination through plaque assays: Using traditional and novel overlay systems. J. Vis. Exp. 2014, 93, e52065. [Google Scholar] [CrossRef]
- Zhang, F.; Chen, Y.; Ke, Y.; Zhang, L.; Zhang, B.; Yang, L.; Zhu, J. Single chain fragment variable (scFv) antibodies targeting the spike protein of porcine epidemic diarrhea virus provide protection against viral infection in piglets. Viruses 2019, 11, 58. [Google Scholar] [CrossRef] [Green Version]
- Waterhouse, A.M.; Procter, J.B.; Martin, D.M.; Clamp, M.; Barton, G.J. Jalview version 2—a multiple sequence alignment editor and analysis workbench. Bioinformatics 2009, 25, 1189–1191. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Winarski, K.L.; Tang, J.; Klenow, L.; Lee, J.; Coyle, E.M.; Manischewitz, J.; Turner, H.L.; Takeda, K.; Ward, A.B.; Golding, H.; et al. Antibody-dependent enhancement of influenza disease promoted by increase in hemagglutinin stem flexibility and virus fusion kinetics. Proc. Natl. Acad. Sci. USA 2019, 116, 15194–15199. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Beam, A.; Goede, D.; Fox, A.; McCool, M.J.; Wall, G.; Haley, C.; Morrison, R. A porcine epidemic diarrhea virus outbreak in one geographic region of the United States: Descriptive epidemiology and investigation of the possibility of airborne virus spread. PLoS ONE 2015, 10, e0144818. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mai, T.N.; Bui, T.P.; Huynh, T.M.L.; Sasaki, Y.; Mitoma, S.; Daous, H.E.; Fahkrajang, W.; Norimine, J.; Sekiguchi, S. Evaluating the risk factors for porcine epidemic diarrhea virus infection in an endemic area of Vietnam. Front. Vet. Sci. 2020, 7, 433. [Google Scholar] [CrossRef] [PubMed]
- Lee, S.H.; Yang, D.-K.; Kim, H.-H.; Cho, I.-S. Efficacy of inactivated variant porcine epidemic diarrhea virus vaccines in growing pigs. Clin. Exp. Vaccine Res. 2018, 7, 61–69. [Google Scholar] [CrossRef]
- Opriessnig, T.; Gerber, P.F.; Shen, H.; de Castro, A.M.; Zhang, J.; Chen, Q.; Halbur, P. Evaluation of the efficacy of a commercial inactivated genogroup 2B-based porcine epidemic diarrhea virus (PEDV) vaccine and experimental live genogroup 1B exposure against 2B challenge. Vet. Res. 2017, 48, 69. [Google Scholar] [CrossRef]
- Bae, J.L.; Lee, J.G.; Kang, T.J.; Jang, H.S.; Jang, Y.S.; Yang, M.S. Induction of antigen-specific systemic and mucosal immune responses by feeding animals transgenic plants expressing the antigen. Vaccine 2003, 21, 4052–4058. [Google Scholar] [CrossRef]
- Wang, C.; Yan, F.; Zheng, X.; Wang, H.; Jin, H.; Wang, C.; Zhao, Y.; Feng, N.; Wang, T.; Gao, Y.; et al. Porcine epidemic diarrhea virus virus-like particles produced in insect cells induce specific immune responses in mice. Virus Genes 2017, 53, 548–554. [Google Scholar] [CrossRef]
- Zhang, Y.; Zhang, X.; Liao, X.; Huang, X.; Cao, S.; Wen, X.; Wen, Y.; Wu, R.; Liu, W. Construction of a bivalent DNA vaccine co-expressing S genes of transmissible gastroenteritis virus and porcine epidemic diarrhea virus delivered by attenuated Salmonella Typhimurium. Virus Genes 2016, 52, 354–364. [Google Scholar] [CrossRef] [PubMed]
- Palatnik-de-Sousa, C.B.; Soares, I.S.; Rosa, D.S. Epitope discovery and synthetic vaccine design. Front. Immunol. 2018, 9, 826. [Google Scholar] [CrossRef] [Green Version]
- Gao, Q.; Zheng, Z.; Wang, H.; Yi, S.; Zhang, G.; Gong, L. The new porcine epidemic diarrhea virus outbreak may mean that existing commercial vaccines are not enough to fully protect against the epidemic strains. Front. Vet. Sci. 2021, 8, 697839. [Google Scholar] [CrossRef] [PubMed]
- Li, R.; Qiao, S.; Yang, Y.; Guo, J.; Xie, S.; Zhou, E.; Zhang, G. Genome sequencing and analysis of a novel recombinant porcine epidemic diarrhea virus strain from Henan, China. Virus Genes 2016, 52, 91–98. [Google Scholar] [CrossRef]
- Yu, J.; Sreenivasan, C.; Uprety, T.; Gao, R.; Huang, C.; Lee, E.J.; Lawson, S.; Nelson, J.; Christopher-Hennings, J.; Kaushik, R.S.; et al. Piglet immunization with a spike subunit vaccine enhances disease by porcine epidemic diarrhea virus. NPJ Vaccines 2021, 6, 22. [Google Scholar] [CrossRef]
- Kao, D.J.; Hodges, R.S. Advantages of a synthetic peptide immunogen over a protein immunogen in the development of an anti-pilus vaccine for Pseudomonas aeruginosa. Chem. Biol. Drug Des. 2009, 74, 33–42. [Google Scholar] [CrossRef] [Green Version]
- Li, F. Structure, function, and evolution of coronavirus Spike proteins. Annu. Rev. Virol. 2016, 3, 237–261. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Shulla, A.; Gallagher, T. Role of spike protein endodomains in regulating coronavirus entry. J. Biol. Chem. 2009, 284, 32725–32734. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bottazzi, M.E.; Strych, U.; Hotez, P.J.; Corry, D.B. Coronavirus vaccine-associated lung immunopathology-what is the significance? Microbes Infect. 2020, 22, 403–404. [Google Scholar] [CrossRef] [PubMed]
- Narayan, R.; Tripathi, S. Intrinsic ADE: The dark side of antibody dependent enhancement during dengue infection. Front. Cell Infect. Microbiol. 2020, 10, 580096. [Google Scholar] [CrossRef]
- Ricke, D.O. Two different antibody-dependent enhancement (ADE) risks for SARS-CoV-2 antibodies. Front. Immunol. 2021, 12, 640093. [Google Scholar] [CrossRef] [PubMed]
- Sánchez-Zuno, G.A.; Matuz-Flores, M.G.; González-Estevez, G.; Nicoletti, F.; Turrubiates-Hernández, F.J.; Mangano, K.; Muñoz-Valle, J.F. A review: Antibody-dependent enhancement in COVID-19: The not so friendly side of antibodies. Int. J. Immunopathol. Pharmacol. 2021, 35, 20587384211050199. [Google Scholar] [CrossRef] [PubMed]
- Pedersen, N.C. An update on feline infectious peritonitis: Virology and immunopathogenesis. Vet. J. 2014, 201, 123–132. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wang, S.F.; Tseng, S.P.; Yen, C.H.; Yang, J.Y.; Tsao, C.H.; Shen, C.W.; Chen, K.H.; Liu, F.T.; Liu, W.T.; Chen, Y.M.; et al. Antibody-dependent SARS coronavirus infection is mediated by antibodies against spike proteins. Biochem. Biophys. Res. Commun. 2014, 451, 208–214. [Google Scholar] [CrossRef] [PubMed]
- Cruz, D.J.; Kim, C.-J.; Shin, H.-J. The GPRLQPY motif located at the carboxy-terminal of the spike protein induces antibodies that neutralize porcine epidemic diarrhea virus. Virus Res. 2008, 132, 192–196. [Google Scholar] [CrossRef]
- Chang, C.Y.; Wang, Y.-S.; Wu, J.-F.; Yang, T.-J.; Chang, Y.-C.; Chae, C.; Chang, H.-W.; Hsu, S.D. Generation and characterization of a spike glycoprotein domain A-specific neutralizing single-chain variable fragment against porcine epidemic diarrhea virus. Vaccines 2021, 9, 833. [Google Scholar] [CrossRef] [PubMed]
- Tien, N.-Q.-D.; Yang, M.-S.; Jang, Y.-S.; Kwon, T.-H.; Reljic, R.; Kim, M.Y. Systemic and oral immunogenicity of porcine epidemic diarrhea virus antigen fused to poly-FC of immunoglobulin G and expressed in ΔXT/ft Nicotiana benthamiana plants. Front. Pharmacol. 2021, 12, 653064. [Google Scholar] [CrossRef]
- Ho, T.T.; Nguyen, G.T.; Pham, N.B.; Le, V.P.; Trinh, T.B.; Vu, T.H.; Phan, H.T.; Conrad, U.; Chu, H.H. Plant-derived trimeric co-26k-equivalent epitope induced neutralizing antibodies against porcine epidemic diarrhea virus. Front. Immunol. 2020, 11, 2152. [Google Scholar] [CrossRef]
- Wang, H.; Zhang, L.; Shang, Y.; Tan, R.; Ji, M.; Yue, X.; Wang, N.; Liu, J.; Wang, C.; Li, Y.; et al. Emergence and evolution of highly pathogenic porcine epidemic diarrhea virus by natural recombination of a low pathogenic vaccine isolate and a highly pathogenic strain in the spike gene. Virus Evol. 2020, 6, veaa049. [Google Scholar] [CrossRef]
- Puente, H.; Argüello, H.; Mencía-Ares, Ó.; Gómez-García, M.; Rubio, P.; Carvajal, A. Detection and genetic diversity of porcine coronavirus involved in diarrhea outbreaks in Spain. Front. Vet. Sci. 2021, 8, 651999. [Google Scholar] [CrossRef]
- Salamat, S.E.; Collantes, T.M.; Lumbera, W.M.; Tablizo, F.A.; Mutia, C.T.; Ong, J.D.; Bandoy, D.J. Sequence analysis of new variants of porcine epidemic diarrhea virus in Luzon, Philippines, in 2017. Arch. Virol. 2021, 166, 1859–1867. [Google Scholar] [CrossRef] [PubMed]
- Li, D.; Li, Y.; Liu, Y.; Chen, Y.; Jiao, W.; Feng, H.; Wei, Q.; Wang, J.; Zhang, Y.; Zhang, G. Isolation and identification of a recombinant porcine epidemic diarrhea virus with a novel insertion in S1 domain. Front. Microbiol. 2021, 12, 667084. [Google Scholar] [CrossRef] [PubMed]
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Thavorasak, T.; Chulanetra, M.; Glab-ampai, K.; Teeranitayatarn, K.; Songserm, T.; Yodsheewan, R.; Sae-lim, N.; Lekcharoensuk, P.; Sookrung, N.; Chaicumpa, W. Novel Neutralizing Epitope of PEDV S1 Protein Identified by IgM Monoclonal Antibody. Viruses 2022, 14, 125. https://doi.org/10.3390/v14010125
Thavorasak T, Chulanetra M, Glab-ampai K, Teeranitayatarn K, Songserm T, Yodsheewan R, Sae-lim N, Lekcharoensuk P, Sookrung N, Chaicumpa W. Novel Neutralizing Epitope of PEDV S1 Protein Identified by IgM Monoclonal Antibody. Viruses. 2022; 14(1):125. https://doi.org/10.3390/v14010125
Chicago/Turabian StyleThavorasak, Techit, Monrat Chulanetra, Kittirat Glab-ampai, Karsidete Teeranitayatarn, Thaweesak Songserm, Rungrueang Yodsheewan, Nawannaporn Sae-lim, Porntippa Lekcharoensuk, Nitat Sookrung, and Wanpen Chaicumpa. 2022. "Novel Neutralizing Epitope of PEDV S1 Protein Identified by IgM Monoclonal Antibody" Viruses 14, no. 1: 125. https://doi.org/10.3390/v14010125