Intranasal Administration of RBD Nanoparticles Confers Induction of Mucosal and Systemic Immunity against SARS-CoV-2
Abstract
:1. Introduction
2. Materials and Methods
2.1. UV-Inactivated SARS-CoV-2
2.2. Animals
2.3. Production of the SARS-CoV-2 RBD Antigen
2.4. Preparation and Characterization of RBD Loaded in N,N,N-trimethyl Chitosan Nanoparticles (RBD-TMC NPs)
2.5. In Vivo Immunization and Specimen Collection
2.6. Quantitation of Antibody Titers
2.7. Whole Virion Capture ELISA
2.8. In Vitro Virus Neutralization Assay
2.9. Detection of RBD-Specific IgA Secreting Cells
2.10. Ex Vivo Stimulation of Splenic Lymphocyte
2.11. Statistical Analysis
3. Results
3.1. Characterization of RBD-TMC NPs
3.2. RBD-TMC NPs Induce Mucosal Immunity in the Respiratory Tract
3.3. RBD-Based TMC Nanoparticle Vaccine Augmented Systemic Humoral Responses
3.4. Neutralizing Activity of RBD-Specific Immune Sera against SARS-CoV-2
3.5. The Intranasal Nanoparticle Vaccine Enhanced the Systemic Cell-Mediated Immune Response
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- V’Kovski, P.; Kratzel, A.; Steiner, S.; Stalder, H.; Thiel, V. Coronavirus biology and replication: Implications for SARS-CoV-2. Nat. Rev. Microbiol. 2021, 19, 155–170. [Google Scholar] [CrossRef]
- Tsatsakis, A.; Calina, D.; Falzone, L.; Petrakis, D.; Mitrut, R.; Siokas, V.; Pennisi, M.; Lanza, G.; Libra, M.; Doukas, S.G.; et al. SARS-CoV-2 pathophysiology and its clinical implications: An integrative overview of the pharmacotherapeutic management of COVID-19. Food. Chem. Toxicol. 2020, 14, 111769. [Google Scholar] [CrossRef]
- Zhou, F.; Yu, T.; Du, R.; Fan, G.; Liu, Y.; Liu, Z.; Xiang, J.; Wang, Y.; Song, B.; Gu, X.; et al. Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: A retrospective cohort study. Lancet 2020, 395, 1054–1062. [Google Scholar] [CrossRef]
- Cuervo, N.Z.; Grandvaux, N. ACE2: Evidence of role as entry receptor for SARS-CoV-2 and implications in comorbidities. Elife 2020, 9, e61390. [Google Scholar] [CrossRef]
- Hoffmann, M.; Kleine-Weber, H.; Schroeder, S.; Krüger, N.; Herrler, T.; Erichsen, S.; Schiergens, T.S.; Herrler, G.; Wu, N.H.; Nitsche, A.; et al. SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. Cell 2020, 181, 271–280. [Google Scholar] [CrossRef] [PubMed]
- Bestle, D.; Heindl, M.R.; Limburg, H.; Van Lam van, T.; Pilgram, O.; Moulton, H.; Stein, D.A.; Hardes, K.; Eickmann, M.; Dolnik, O.; et al. TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells. Life Sci. Alliance 2020, 3, e202000786. [Google Scholar] [CrossRef]
- Walls, A.C.; Park, Y.J.; Tortorici, M.A.; Wall, A.; McGuire, A.T.; Veesler, D. Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein. Cell 2020, 181, 281–292. [Google Scholar] [CrossRef]
- Xia, S.; Zhu, Y.; Liu, M.; Lan, Q.; Xu, W.; Wu, Y.; Ying, T.; Liu, S.; Shi, Z.; Jiang, S.; et al. Fusion mechanism of 2019-nCoV and fusion inhibitors targeting HR1 domain in spike protein. Cell Mol. Immunol. 2020, 17, 765–767. [Google Scholar] [CrossRef]
- Poh, C.M.; Carissimo, G.; Wang, B.; Amrun, S.N.; Lee, C.Y.; Chee, R.S.; Fong, S.W.; Yeo, N.K.; Lee, W.H.; Torres-Ruesta, A.; et al. Two linear epitopes on the SARS-CoV-2 spike protein that elicit neutralising antibodies in COVID-19 patients. Nat. Commun. 2020, 11, 2806. [Google Scholar] [CrossRef]
- Lin, L.; Ting, S.; Yufei, H.; Wendong, L.; Yubo, F.; Jing, Z. Epitope-based peptide vaccines predicted against novel coronavirus disease caused by SARS-CoV-2. Virus Res. 2020, 288, 198082. [Google Scholar] [CrossRef]
- Muraoka, D.; Situo, D.; Sawada, S.I.; Akiyoshi, K.; Harada, N.; Ikeda, H. Identification of a dominant CD8(+) CTL epitope in the SARS-associated coronavirus 2 spike protein. Vaccine 2020, 38, 7697–7701. [Google Scholar] [CrossRef] [PubMed]
- Huang, P.H.; Tsai, H.H.; Liao, B.H.; Lin, Y.L.; Jan, J.T.; Tao, M.H.; Chou, Y.C.; Hu, C.J.; Chen, H.W. Neutralizing antibody response elicited by SARS-CoV-2 receptor-binding domain. Hum. Vaccines Immunother. 2021, 17, 654–655. [Google Scholar] [CrossRef] [PubMed]
- Sato, S.; Kiyono, H. The mucosal immune system of the respiratory tract. Curr. Opin. Virol. 2012, 2, 225–232. [Google Scholar] [CrossRef] [PubMed]
- Brandtzaeg, P. Chapter 103—Immunobiology of the Tonsils and Adenoids. In Mucosal Immunology, 4th ed.; Mestecky, J., Strober, W., Russell, M.W., Kelsall, B.L., Cheroutre, H., Lambrecht, B.N., Eds.; Academic Press: Boston, MA, USA, 2015; pp. 1985–2016. [Google Scholar]
- Marasini, N.; Skwarczynski, M.; Toth, I. Intranasal delivery of nanoparticle-based vaccines. Ther. Deliv. 2017, 8, 151–167. [Google Scholar] [CrossRef] [PubMed]
- Pati, R.; Shevtsov, M.; Sonawane, A. Nanoparticle Vaccines Against Infectious Diseases. Front. Immunol. 2018, 9, 2224. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dabaghian, M.; Latifi, A.M.; Tebianian, M.; NajmiNejad, H.; Ebrahimi, S.M. Nasal vaccination with r4M2e.HSP70c antigen encapsulated into N-trimethyl chitosan (TMC) nanoparticulate systems: Preparation and immunogenicity in a mouse model. Vaccine 2018, 36, 2886–2895. [Google Scholar] [CrossRef]
- Najminejad, H.; Kalantar, S.M.; Mokarram, A.R.; Dabaghian, M.; Abdollahpour-Alitappeh, M.; Ebrahimi, S.M.; Tebianian, M.; Ramandi, M.F.; Sheikhha, M.H. Bordetella pertussis antigens encapsulated into N-trimethyl chitosan nanoparticulate systems as a novel intranasal pertussis vaccine. Artif. Cells Nanomed. Biotechnol. 2019, 47, 2605–2611. [Google Scholar] [CrossRef] [PubMed]
- Shim, S.; Yoo, H.S. The Application of Mucoadhesive Chitosan Nanoparticles in Nasal Drug Delivery. Mar. Drugs 2020, 18, 605. [Google Scholar] [CrossRef]
- Zhang, P.; Liu, W.; Peng, Y.; Han, B.; Yang, Y. Toll like receptor 4 (TLR4) mediates the stimulating activities of chitosan oligosaccharide on macrophages. Int. Immunopharmacol. 2014, 23, 254–261. [Google Scholar] [CrossRef]
- Walter, F.; Winter, E.; Rahn, S.; Heidland, J.; Meier, S.; Struzek, A.M.; Lettau, M.; Philipp, L.M.; Beckinger, S.; Otto, L.; et al. Chitosan nanoparticles as antigen vehicles to induce effective tumor specific T cell responses. PLoS ONE 2020, 15, e0239369. [Google Scholar] [CrossRef]
- Rungrojcharoenkit, K.; Sunintaboon, P.; Ellison, D.; Macareo, L.; Midoeng, P.; Chaisuwirat, P.; Fernandez, S.; Ubol, S. Development of an adjuvanted nanoparticle vaccine against influenza virus, an in vitro study. PLoS ONE 2020, 15, e0237218. [Google Scholar] [CrossRef]
- Jearanaiwitayakul, T.; Sunintaboon, P.; Chawengkittikul, R.; Limthongkul, J.; Midoeng, P.; Warit, S.; Ubol, S. Nanodelivery system enhances the immunogenicity of dengue-2 nonstructural protein 1, DENV-2 NS1. Vaccine 2020, 38, 6814–6825. [Google Scholar] [CrossRef]
- Shah, H.B.; Koelsch, K.A. B-Cell ELISPOT: For the Identification of Antigen-Specific Antibody-Secreting Cells. Methods Mol. Biol. 2015, 1312, 419–426. [Google Scholar] [CrossRef]
- Cervia, C.; Nilsson, J.; Zurbuchen, Y.; Valaperti, A.; Schreiner, J.; Wolfensberger, A.; Raeber, M.E.; Adamo, S.; Weigang, S.; Emmenegger, M.; et al. Systemic and mucosal antibody responses specific to SARS-CoV-2 during mild versus severe COVID-19. J. Allergy Clin. Immunol. 2021, 147, 545–557. [Google Scholar] [CrossRef]
- Sterlin, D.; Mathian, A.; Miyara, M.; Mohr, A.; Anna, F.; Claër, L.; Quentric, P.; Fadlallah, J.; Devilliers, H.; Ghillani, P.; et al. IgA dominates the early neutralizing antibody response to SARS-CoV-2. Sci. Transl. Med. 2021, 13, eabd2223. [Google Scholar] [CrossRef] [PubMed]
- Wang, Z.; Lorenzi, J.C.C.; Muecksch, F.; Finkin, S.; Viant, C.; Gaebler, C.; Cipolla, M.; Hoffmann, H.H.; Oliveira, T.Y.; Oren, D.A.; et al. Enhanced SARS-CoV-2 neutralization by dimeric IgA. Sci. Transl. Med. 2021, 13, eabf1555. [Google Scholar] [CrossRef]
- Firacative, C.; Gressler, A.E.; Schubert, K.; Schulze, B.; Müller, U.; Brombacher, F.; von Bergen, M.; Alber, G. Identification of T helper (Th)1- and Th2-associated antigens of Cryptococcus neoformans in a murine model of pulmonary infection. Sci. Rep. 2018, 8, 2681. [Google Scholar] [CrossRef] [PubMed]
- Cêtre, C.; Pierrot, C.; Cocude, C.; Lafitte, S.; Capron, A.; Capron, M.; Khalife, J. Profiles of Th1 and Th2 cytokines after primary and secondary infection by Schistosoma mansoni in the semipermissive rat host. Infect. Immun. 1999, 67, 2713–2719. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chang, S.Y.; Ko, H.J.; Kweon, M.N. Mucosal dendritic cells shape mucosal immunity. Exp. Mol. Med. 2014, 46, e84. [Google Scholar] [CrossRef] [PubMed]
- Pabst, R. Mucosal vaccination by the intranasal route. Nose-associated lymphoid tissue (NALT)-Structure, function and species differences. Vaccine 2015, 33, 4406–4413. [Google Scholar] [CrossRef]
- Hassan, A.O.; Kafai, N.M.; Dmitriev, I.P.; Fox, J.M.; Smith, B.K.; Harvey, I.B.; Chen, R.E.; Winkler, E.S.; Wessel, A.W.; Case, J.B.; et al. A Single-Dose Intranasal ChAd Vaccine Protects Upper and Lower Respiratory Tracts against SARS-CoV-2. Cell 2020, 183, 169–184. [Google Scholar] [CrossRef] [PubMed]
- Sperandio, B.; Fischer, N.; Sansonetti, P.J. Mucosal physical and chemical innate barriers: Lessons from microbial evasion strategies. Semin. Immunol. 2015, 27, 111–118. [Google Scholar] [CrossRef]
- Foged, C.; Brodin, B.; Frokjaer, S.; Sundblad, A. Particle size and surface charge affect particle uptake by human dendritic cells in an in vitro model. Int. J. Pharm. 2005, 298, 315–322. [Google Scholar] [CrossRef]
- Mohammed, M.A.; Syeda, J.T.M.; Wasan, K.M.; Wasan, E.K. An Overview of Chitosan Nanoparticles and Its Application in Non-Parenteral Drug Delivery. Pharmaceutics 2017, 9, 53. [Google Scholar] [CrossRef] [Green Version]
- Liu, Q.; Zheng, X.; Zhang, C.; Shao, X.; Zhang, X.; Zhang, Q.; Jiang, X. Conjugating influenza a (H1N1) antigen to n-trimethylaminoethylmethacrylate chitosan nanoparticles improves the immunogenicity of the antigen after nasal administration. J. Med. Virol. 2015, 87, 1807–1815. [Google Scholar] [CrossRef]
- Slütter, B.; Bal, S.; Keijzer, C.; Mallants, R.; Hagenaars, N.; Que, I.; Kaijzel, E.; van Eden, W.; Augustijns, P.; Löwik, C.; et al. Nasal vaccination with N-trimethyl chitosan and PLGA based nanoparticles: Nanoparticle characteristics determine quality and strength of the antibody response in mice against the encapsulated antigen. Vaccine 2010, 28, 6282–6291. [Google Scholar] [CrossRef] [PubMed]
- Liew, F.Y.; Russell, S.M.; Appleyard, G.; Brand, C.M.; Beale, J. Cross-protection in mice infected with influenza A virus by the respiratory route is correlated with local IgA antibody rather than serum antibody or cytotoxic T cell reactivity. Eur. J. Immunol. 1984, 14, 350–356. [Google Scholar] [CrossRef] [PubMed]
- Li, Y.; Jin, L.; Chen, T. The Effects of Secretory IgA in the Mucosal Immune System. Biomed. Res. Int. 2020, 2020, 2032057. [Google Scholar] [CrossRef]
- Bidgood, S.R.; Tam, J.C.; McEwan, W.A.; Mallery, D.L.; James, L.C. Translocalized IgA mediates neutralization and stimulates innate immunity inside infected cells. Proc. Natl. Acad. Sci. USA 2014, 111, 13463–13468. [Google Scholar] [CrossRef] [Green Version]
- Abkar, M.; Fasihi-Ramandi, M.; Kooshki, H.; Lotfi, A.S. Oral immunization of mice with Omp31-loaded N-trimethyl chitosan nanoparticles induces high protection against Brucella melitensis infection. Int. J. Nanomed. 2017, 12, 8769–8778. [Google Scholar] [CrossRef] [Green Version]
- Amidi, M.; Romeijn, S.G.; Verhoef, J.C.; Junginger, H.E.; Bungener, L.; Huckriede, A.; Crommelin, D.J.; Jiskoot, W. N-trimethyl chitosan (TMC) nanoparticles loaded with influenza subunit antigen for intranasal vaccination: Biological properties and immunogenicity in a mouse model. Vaccine 2007, 25, 144–153. [Google Scholar] [CrossRef]
- Guthmiller, J.J.; Stovicek, O.; Wang, J.; Changrob, S.; Li, L.; Halfmann, P.; Zheng, N.-Y.; Utset, H.; Stamper, C.T.; Dugan, H.L.; et al. SARS-CoV-2 Infection Severity Is Linked to Superior Humoral Immunity against the Spike. mBio 2021, 12, e02940-20. [Google Scholar] [CrossRef]
- Ren, L.; Zhang, L.; Chang, D.; Wang, J.; Hu, Y.; Chen, H.; Guo, L.; Wu, C.; Wang, C.; Wang, Y.; et al. The kinetics of humoral response and its relationship with the disease severity in COVID-19. Commun. Biol. 2020, 3, 780. [Google Scholar] [CrossRef] [PubMed]
- Seow, J.; Graham, C.; Merrick, B.; Acors, S.; Pickering, S.; Steel, K.J.A.; Hemmings, O.; O’Byrne, A.; Kouphou, N.; Galao, R.P.; et al. Longitudinal observation and decline of neutralizing antibody responses in the three months following SARS-CoV-2 infection in humans. Nat. Microbiol. 2020, 5, 1598–1607. [Google Scholar] [CrossRef]
- Bilich, T.; Nelde, A.; Heitmann, J.S.; Maringer, Y.; Roerden, M.; Bauer, J.; Rieth, J.; Wacker, M.; Hörber, S.; Rachfalski, D.; et al. T cell and antibody kinetics delineate SARS-CoV-2 peptides mediating long-term immune responses in COVID-19 convalescent individuals. Sci. Transl. Med. 2021, 13, eabf7517. [Google Scholar] [CrossRef]
- Peng, Y.; Mentzer, A.J.; Liu, G.; Yao, X.; Yin, Z.; Dong, D.; Dejnirattisai, W.; Rostron, T.; Supasa, P.; Liu, C.; et al. Broad and strong memory CD4+ and CD8+ T cells induced by SARS-CoV-2 in UK convalescent individuals following COVID-19. Nat. Immunol. 2020, 21, 1336–1345. [Google Scholar] [CrossRef] [PubMed]
- Zhang, B.-Z.; Hu, Y.-F.; Chen, L.-L.; Yau, T.; Tong, Y.-G.; Hu, J.-C.; Cai, J.-P.; Chan, K.-H.; Dou, Y.; Deng, J.; et al. Mining of epitopes on spike protein of SARS-CoV-2 from COVID-19 patients. Cell Res. 2020, 30, 702–704. [Google Scholar] [CrossRef] [PubMed]
- Smith, S.A.; Selby, L.I.; Johnston, A.P.R.; Such, G.K. The Endosomal Escape of Nanoparticles: Toward More Efficient Cellular Delivery. Bioconjug. Chem. 2019, 30, 263–272. [Google Scholar] [CrossRef] [PubMed]
Nanoparticles | Particle Size (nm) | Polydispersity Index (PDI) | Zeta Potential (mV) | % Loading Efficiency (LE) |
---|---|---|---|---|
TMC NPs | 380.3 ± 15.11 | 0.410 ± 0.028 | 16.0 ± 0.208 | - |
RBD-TMC NPs | 386.5 ± 58.96 | 0.407 ± 0.019 | 12.9 ± 0.651 | 99.32 ± 1.18 |
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
Jearanaiwitayakul, T.; Seesen, M.; Chawengkirttikul, R.; Limthongkul, J.; Apichirapokey, S.; Sapsutthipas, S.; Phumiamorn, S.; Sunintaboon, P.; Ubol, S. Intranasal Administration of RBD Nanoparticles Confers Induction of Mucosal and Systemic Immunity against SARS-CoV-2. Vaccines 2021, 9, 768. https://doi.org/10.3390/vaccines9070768
Jearanaiwitayakul T, Seesen M, Chawengkirttikul R, Limthongkul J, Apichirapokey S, Sapsutthipas S, Phumiamorn S, Sunintaboon P, Ubol S. Intranasal Administration of RBD Nanoparticles Confers Induction of Mucosal and Systemic Immunity against SARS-CoV-2. Vaccines. 2021; 9(7):768. https://doi.org/10.3390/vaccines9070768
Chicago/Turabian StyleJearanaiwitayakul, Tuksin, Mathurin Seesen, Runglawan Chawengkirttikul, Jitra Limthongkul, Suttikarn Apichirapokey, Sompong Sapsutthipas, Supaporn Phumiamorn, Panya Sunintaboon, and Sukathida Ubol. 2021. "Intranasal Administration of RBD Nanoparticles Confers Induction of Mucosal and Systemic Immunity against SARS-CoV-2" Vaccines 9, no. 7: 768. https://doi.org/10.3390/vaccines9070768
APA StyleJearanaiwitayakul, T., Seesen, M., Chawengkirttikul, R., Limthongkul, J., Apichirapokey, S., Sapsutthipas, S., Phumiamorn, S., Sunintaboon, P., & Ubol, S. (2021). Intranasal Administration of RBD Nanoparticles Confers Induction of Mucosal and Systemic Immunity against SARS-CoV-2. Vaccines, 9(7), 768. https://doi.org/10.3390/vaccines9070768