An innovative nasal nanovaccine against SARS-CoV-2 induces systemic and upper airway immunity controlling viral replication

an-innovative-nasal-nanovaccine-against-sars-cov-2-induces-systemic-and-upper-airway-immunity-controlling-viral-replication
An innovative nasal nanovaccine against SARS-CoV-2 induces systemic and upper airway immunity controlling viral replication

References

  1. World Health Organization. The top 10 causes of death https://www.who.int/news-room/fact-sheets/detail/the-top-10-causes-of-death (2020).

  2. World Health Organization. COVID-19 vaccine tracker and landscape https://www.who.int/publications/m/item/draft-landscape-of-covid-19-candidate-vaccines (2023).

  3. Shang, J. et al. Structural basis of receptor recognition by SARS-CoV-2. Nature 581, 221–224 (2020).

    Google Scholar 

  4. Wrapp, D. et al. Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science 367, 1260–1263 (2020).

    Google Scholar 

  5. Bourgonje, A. R. et al. Angiotensin-converting enzyme 2 (ACE2), SARS-CoV-2 and the pathophysiology of coronavirus disease 2019 (COVID-19). J. Pathol. 251, 228–248 (2020).

    Google Scholar 

  6. Mohammed, I. et al. The efficacy and effectiveness of the COVID-19 vaccines in reducing infection, severity, hospitalization, and mortality: a systematic review. Hum. Vaccin. Immunother. 18, 2027160 (2022).

    Google Scholar 

  7. Bleier, B. S., Ramanathan, M. & Lane, A. P. COVID-19 vaccines may not prevent nasal SARS-CoV-2 infection and asymptomatic transmission. Otolaryngol. Head. Neck Surg. 164, 305–307 (2020).

    Google Scholar 

  8. Liew, F. et al. SARS-CoV-2-specific nasal IgA wanes 9 months after hospitalisation with COVID-19 and is not induced by subsequent vaccination. EBioMedicine 87, 104402 (2023).

    Google Scholar 

  9. Fröberg, J. et al. SARS-CoV-2 mucosal antibody development and persistence and their relation to viral load and COVID-19 symptoms. Nat. Commun. 12, 5621 (2021).

    Google Scholar 

  10. Pilapitiya, D., Wheatley, A. K. & Tan, H.-X. Mucosal vaccines for SARS-CoV-2: triumph of hope over experience. EBioMedicine 92, 104585 (2023).

    Google Scholar 

  11. Kehagia, E., Papakyriakopoulou, P. & Valsami, G. Advances in intranasal vaccine delivery: a promising non-invasive route of immunization. Vaccine 41, 3589–3603 (2023).

    Google Scholar 

  12. Slomski, A. Intranasal COVID-19 vaccine disappointing in first-in-human trial. JAMA 328, 2003 (2022).

    Google Scholar 

  13. Zhu, F. et al. Safety and efficacy of the intranasal spray SARS-CoV-2 vaccine dNS1-RBD: a multicentre, randomised, double-blind, placebo-controlled, phase 3 trial. Lancet Respir. Med. 11, 1075–1088 (2023).

    Google Scholar 

  14. Sun, B. et al. An intranasally administered adenovirus-vectored SARS-CoV-2 vaccine induces robust mucosal secretory IgA. JCI Insight 9, e180784 (2024).

    Google Scholar 

  15. Ndzouboukou, J. B., Kamara, A. A., Ullah, N., Lei, Q. & Fan, X. L. A Meta-analysis on the immunogenicity of homologous versus heterologous immunization regimens against SARS-CoV-2 Beta, Delta, and Omicron BA.1 VoCs in healthy adults. J. Microbiol. Biotechnol. 35, e2411059 (2025).

    Google Scholar 

  16. Nissilä, E. et al. The COVID-19 vaccine ChAdOx1 is opsonized by anti-vector antibodies that activate complement and promote viral vector phagocytosis. Scand. J. Immunol. 101, e70000 (2025).

    Google Scholar 

  17. Yang, Y., Zhang, M., Song, H. & Yu, C. Silica-based nanoparticles for biomedical applications: from nanocarriers to biomodulators. Acc. Chem. Res. 53, 1545–1556 (2020).

    Google Scholar 

  18. Liljenström, C.L.D.; Finnveden, G. Silicon-Based Nanomaterials in a Life-Cycle Perspective, Including a Case Study on SelfCleaning Coatings https://www.researchgate.net/publication/280264076_Silicon-based_nanomaterials_in_a_life-cycle_perspective_including_a_case_study_on_self-cleaning_coatings (2013).

  19. Tang, L. & Cheng, J. Nonporous silica nanoparticles for nanomedicine application. Nano Today 8, 290–312 (2013).

    Google Scholar 

  20. Janjua, T. I., Cao, Y., Yu, C. & Popat, A. Clinical translation of silica nanoparticles. Nat. Rev. Mater. 6, 1072–1074 (2021).

    Google Scholar 

  21. Tan, A., Eskandar, N. G., Rao, S. & Prestidge, C. A. First in man bioavailability and tolerability studies of a silica–lipid hybrid (Lipoceramic) formulation: a Phase I study with ibuprofen. Drug Deliv. Transl. Res. 4, 212–221 (2014).

    Google Scholar 

  22. Meola, T. R., Abuhelwa, A. Y., Joyce, P., Clifton, P. & Prestidge, C. A. A safety, tolerability, and pharmacokinetic study of a novel simvastatin silica-lipid hybrid formulation in healthy male participants. Drug Deliv. Transl. Res. 11, 1261–1272 (2021).

    Google Scholar 

  23. Barandeh, F. et al. Organically modified silica nanoparticles are biocompatible and can be targeted to neurons in vivo. PLoS ONE 7, e29424 (2012).

    Google Scholar 

  24. Brown, S. C. et al. Influence of shape, adhesion and simulated lung mechanics on amorphous silica nanoparticle toxicity. Adv. Powder Technol. 18, 69–79 (2007).

    Google Scholar 

  25. Fruijtier-Pölloth, C. The toxicological mode of action and the safety of synthetic amorphous silica-a nanostructured material. Toxicology 294, 61–79 (2012).

    Google Scholar 

  26. Yu, T., Greish, K., McGill, L. D., Ray, A. & Ghandehari, H. Influence of geometry, porosity, and surface characteristics of silica nanoparticles on acute toxicity: their vasculature effect and tolerance threshold. ACS Nano 6, 2289–2301 (2012).

    Google Scholar 

  27. European Medicines Agency, Scientific Committee on Consumer Safety opinion on silica, hydrated silica, and silica surface modified with alkyl silicates (nano form) https://ec.europa.eu/health/scientific_committees/consumer_safety/docs/sccs_o_175.pdf (2015).

  28. U.S Food and Drug Administration, US Food and Drug Administration GRAS Substances (SCOGS) Database—Select Committee on GRAS Substances (SCOGS) Opinion: Silicates, (n.d.) https://www.fda.gov/food/generally-recognized-safe-gras/gras-substances-scogs-database (2022).

  29. FDA, 2023 U.S. FDA Drug Master Files (DMF) 1Q2023 https://www.fda.gov/media/166951/download (2023).

  30. EMA, 2017 Cyclodextrins used as excipients. Report published in support of the ‘Questions and answers on cyclodextrins used as excipients in medicinal products for human use’ (EMA/CHMP/495747/2013) https://www.ema.europa.eu/en/documents/scientific-guideline/questions-and-answers-cyclodextrins-used-excipients-medicinal-products-human-use_en.pdf (2017).

  31. Rassu, G. et al. Versatile nasal application of cyclodextrins: excipients and/or actives. Pharmaceutics 13, 1180 (2021).

    Google Scholar 

  32. Marttin, E., Verhoef, J. C. & Merkus, F. W. Efficacy, safety and mechanism of cyclodextrins as absorption enhancers in nasal delivery of peptide and protein drugs. J. Drug Target. 6, 17–36 (1998).

    Google Scholar 

  33. Asim, M. H. et al. S-protected thiolated cyclodextrins as mucoadhesive oligomers for drug delivery. J. Colloid Interface Sci. 531, 261–268 (2018).

    Google Scholar 

  34. Jansook, P., Ogawa, N. & Loftsson, T. Cyclodextrins: structure, physicochemical properties and pharmaceutical applications. Int. J. Pharm. 535, 272–284 (2018).

    Google Scholar 

  35. Haimhoffer, A. et al. Cyclodextrins in drug delivery systems and their effects on biological barriers. Sci. Pharm. 87, 33 (2019).

    Google Scholar 

  36. Matassoli, F. L. et al. Hydroxypropyl-beta-cyclodextrin reduces inflammatory signaling from monocytes: possible implications for suppression of HIV chronic immune activation. mSphere 3, e00497–18 (2018).

    Google Scholar 

  37. Lu, A. et al. Hydroxypropyl-beta cyclodextrin barrier prevents respiratory viral infections: a preclinical study. Int. J. Mol. Sci. 25, 2061 (2024).

    Google Scholar 

  38. Asai, K. et al. The effects of water-soluble cyclodextrins on the histological integrity of the rat nasal mucosa. Int. J. Pharm. 246, 25–35 (2002).

    Google Scholar 

  39. Agu, R. U. et al. Safety assessment of selected cyclodextrin-effect of ciliary activity using a human cell suspension culture model exhibiting in-vitro ciliogenesis. Int. J. Pharm. 193, 219–226 (2000).

    Google Scholar 

  40. Yuan, M., Liu, H., Wu, N. C. & Wilson, I. A. Recognition of the SARS-CoV-2 receptor binding domain by neutralizing antibodies. Biochem. Biophys. Res. Commun. 538, 192–203 (2021).

    Google Scholar 

  41. Fernandes, E. R. et al. Time-dependent contraction of the SARS-CoV-2-specific T-cell responses in convalescent individuals. JACI Glob. 1, 112–121 (2022).

    Google Scholar 

  42. Ichinohe, T. et al. Synthetic double-stranded RNA poly(I:C) combined with mucosal vaccine protects against influenza virus infection. J. Virol. 79, 2910–2919 (2005).

    Google Scholar 

  43. Sloat, B. R. & Cui, Z. Nasal immunization with anthrax protective antigen protein adjuvanted with polyriboinosinic-polyribocytidylic acid-induced strong mucosal and systemic immunities. Pharm. Res. 23, 1217–1226 (2006).

    Google Scholar 

  44. Netsomboon, K. & Bernkop-Schnürch, A. Mucoadhesive vs. mucopenetrating particulate drug delivery. Eur. J. Pharm. Biopharm. 98, 76–89 (2016).

    Google Scholar 

  45. Corr, S. C., Gahan, C. C. G. M. & Hill, C. M-cells: origin, morphology and role in mucosal immunity and microbial pathogenesis. FEMS Immunol. Med. Microbiol. 52, 2–12 (2008).

    Google Scholar 

  46. Kiyono, H. & Fukuyama, S. NALT- versus Peyer’s-patch-mediated mucosal immunity. Nat. Rev. Immunol. 4, 699–710 (2004).

    Google Scholar 

  47. Gómez, D. M., Urcuqui-Inchima, S. & Hernandez, J. C. Silica nanoparticles induce NLRP3 inflammasome activation in human primary immune cells. Innate Immun. 23, 697–708 (2017).

    Google Scholar 

  48. Tang, L., Fan, T. M., Borst, L. B. & Cheng, J. Synthesis and biological response of size-specific, monodisperse drug-silica nanoconjugates. ACS Nano 6, 3954–3966 (2012).

    Google Scholar 

  49. Hirai, T. et al. Amorphous silica nanoparticles enhance cross-presentation in murine dendritic cells. Biochem. Biophys. Res. Commun. 427, 553–556 (2012).

    Google Scholar 

  50. Vis, B. et al. Non-functionalized ultrasmall silica nanoparticles directly and size-selectively activate T cells. ACS Nano 12, 10843–10854 (2018).

    Google Scholar 

  51. Onishi, M. et al. Hydroxypropyl-beta-cyclodextrin spikes local inflammation that induces Th2 cell and T follicular helper cell responses to the coadministered antigen. J. Immunol. 194, 2673–2682 (2015).

    Google Scholar 

  52. Sanità, G., Carrese, B. & Lamberti, A. Nanoparticle surface functionalization: how to improve biocompatibility and cellular internalization. Front. Mol. Biosci. 7, 587012 (2020).

    Google Scholar 

  53. Button, B. et al. A periciliary brush promotes the lung health by separating the mucus layer from airway epithelia. Science 337, 937–941 (2012).

    Google Scholar 

  54. de Steenhuijsen Piters, W. A. A. et al. Interaction between the nasal microbiota and S. pneumoniae in the context of live-attenuated influenza vaccine. Nat. Commun. 10, 2981 (2019). 9.

    Google Scholar 

  55. Liu, X., Wetzler, L. M., Nascimento, L. O. & Massari, P. Human airway epithelial cell responses to Neisseria lactamica and purified porin via Toll-like receptor 2-dependent signaling. Infect. Immun. 78, 5314–5323 (2010).

    Google Scholar 

  56. de Fays, C., Carlier, F. M., Gohy, S. & Pilette, C. Secretory immunoglobulin A immunity in chronic obstructive respiratory diseases. Cells 11, 1324 (2022).

    Google Scholar 

  57. Firacative, C. et al. Identification of T helper (Th)1- and Th2-associated antigens of Cryptococcus neoformans in a murine model of pulmonary infection. Sci. Rep. 8, 2681 (2018).

    Google Scholar 

  58. Khoury, D. S. et al. Measuring immunity to SARS-CoV-2 infection: comparing assays and animal models. Nat. Rev. Immunol. 20, 727–738; 1 (2020).

    Google Scholar 

  59. Liu, C., Jiang, X., Gan, Y. & Yu, M. Engineering nanoparticles to overcome the mucus barrier for drug delivery: design, evaluation and state-of-the-art. Med. Drug Discov. 12, 100110 (2021).

    Google Scholar 

  60. Honary, S. & Zahir, F. Effect of zeta potential on the properties of nano-drug delivery systems – A review (Part 1). Trop. J. Pharm. Res. 12, 255–264 (2013).

    Google Scholar 

  61. Foroozandeh, P. & Aziz, A. A. Insight into cellular uptake and intracellular trafficking of nanoparticles. Nanoscale Res. Lett. 13, 339 (2018).

    Google Scholar 

  62. Newby, J. M. et al. Technologies strategies to estimate and control diffuse passage times through the mucus barrier in mucosal drug delivery. Adv. Drug Deliv. Rev. 124, 64–81 (2018).

    Google Scholar 

  63. Li, M. et al. Enhanced intranasal delivery of mRNA vaccine by overcoming the nasal epithelial barrier via intra- and paracellular pathways. J. Control. Release 228, 9–19 (2016).

    Google Scholar 

  64. Li, L. et al. Biodistribution, excretion, and toxicity of mesoporous silica nanoparticles after oral administration depend on their shape. Nanomedicine 11, 1915–1924 (2015).

    Google Scholar 

  65. He, X. X. et al. In vivo study of biodistribution and urinary excretion of surface-modified silica nanoparticles. Analy. Chem. 80, 9597–9603 (2008).

    Google Scholar 

  66. Choi, H. S. et al. Renal clearance of nanoparticles. Nat. Biotechnol. 25, 1165–1170 (2009).

    Google Scholar 

  67. Arick, D. Q., Choi, Y. H., Kim, H. C. & Won, Y.-Y. Effects of nanoparticles on the mechanical functioning of the lung. Adv. Colloid Interface Sci. 225, 218–228 (2015).

    Google Scholar 

  68. Gu, X. et al. Clearance of two organic nanoparticles from the brain via the paravascular pathway. J. Control. Rel. 322, 31–41 (2020).

    Google Scholar 

  69. Formica, M. L. et al. On a highway to the brain: a review on nose-to-brain drug delivery using nanoparticles. Appl. Mater. Today 29, 101631 (2022).

    Google Scholar 

  70. Hajdu, I. et al. Radiochemical synthesis and preclinical evaluation of 68Ga-labeled NODAGA-hydroxypropyl-beta-cyclodextrin (68Ga-NODAGA-HPBCD). Eur. J. Pharm. Sci. 128, 202–208 (2019).

    Google Scholar 

  71. Carter, N. J. & Curran, M. P. Live attenuated influenza vaccine (Flumist; Fluenz): a review of its use in prevention of seasonal influenza in children and adults. Drugs 71, 1591–1622 (2011).

    Google Scholar 

  72. Mutsch, M. et al. Use of the inactivated intranasal influenza vaccine and the risk of Bell’s palsy in Switzerland. N. Engl. J. Med. 350, 896–903 (2004).

    Google Scholar 

  73. Madhavan, M. et al. Tolerability and immunogenicity of an intranasally-administered adenovirus-vectored COVID-19 vaccine: an open-label partially-randomised ascending dose phase I trial. EBioMedicine 85, 104298 (2022).

    Google Scholar 

  74. Knudsen, K. B. et al. In vivo toxicity of cationic micelles and liposomes. Nanomedicine 11, 467–477 (2015).

    Google Scholar 

  75. Zhang, P. et al. Nanoparticle size influences antigen retention and presentation in lymph node. ACS Nano 9, 8480–8493 (2015).

    Google Scholar 

  76. Southam, D. S., Dolovich, M., O’Byrne, P. M. & Inman, M. D. Distribution of intranasal instillations in mice: effects of volume, time, body position, and anesthesia. Am. J. Physiol. Lung Cell Mol. Physiol. 282, L833–L839 (2002).

    Google Scholar 

  77. Mitchell, M. J. et al. Engineering precision nanoparticles for drug delivery. Nat. Rev. Drug Discov. 20, 101–124 (2021).

    Google Scholar 

  78. Corthésy, B. Role of secretory IgA in infection and immunity. Mucosal Immunol. 6, 535–542 (2013).

    Google Scholar 

  79. Wright, P. F. et al. Longitudinal systemic and mucosal immune response to SARS-CoV-2 infection. J. Infect. Dis. 226, 1204–1214 (2022).

    Google Scholar 

  80. Hartwell, B. L. et al. Intranasal vaccination with lipid-conjugated immunogens promotes antigen transmucosal uptake to drive mucosal and systemic immunity. Sci. Transl. Med. 14, eabn1413 (2022).

    Google Scholar 

  81. Adhikari, K. & Verma, S. C. Neutralization antibody responses to SARS-CoV-2 variants after COVID-19 vaccination and boosters. Vaccine 24, 100664 (2025).

    Google Scholar 

  82. Bladh, O. et al. Comparison of SARS-CoV-2 spike-specific IgA and IgG in nasal secretions, saliva and serum. Front. Immunol. 15, 1346749 (2024).

    Google Scholar 

  83. Riepler, L. et al. Comparison of four SARS-CoV-2 neutralization assays. Vaccines 9, 13 (2020).

    Google Scholar 

  84. Klein, S. L. & Flanagan, K. L. Sex differences in immune responses. Nat. Rev. Immunol. 16, 626–638 (2016).

    Google Scholar 

  85. Huang, L.-Y., Stuart, C., Takeda, K., D’Agnillo, F. & Golding, B. Poly(I:C) induces human lung endothelial barrier dysfunction by disrupting tight junction expression of claudin-5. PloS One 11, e0160875 (2016).

    Google Scholar 

  86. Starkhammar, M. et al. Intranasal administration of poly(I:C) and LPS in BALB/c mice induces airway hyperresponsiveness and inflammation via different pathways. PLoS ONE 7, e32110 (2012).

    Google Scholar 

  87. Misra, S. K., Kapoor, A. & Pathak, K. Nanovaccines for Mucosal Immunity (eds Chavda, V. P. & Apostolopoulos, V.) 367–404 (Wiley online library, 2024).

  88. WHO. Recommendations to assure the quality, safety and efficacy of influenza vaccines (human, live attenuated) for intranasal administration, Annex 4, WHO Technical Report Series No. 977 https://www.who.int/publications/m/item/influenza-attenuated-intranasal-administration-annex-4-trs-no-977 (2013).

  89. Nair, A. B. & Jacob, S. A simple practice guide for dose conversion between animals and human. J. Basic Clin. Pharm. 7, 27–31 (2016).

    Google Scholar 

  90. Đorđević, S. et al. Current hurdles to the translation of nanomedicines from bench to the clinic. Drug Deliv. Transl. Res. 12, 500–525; 2 (2022).

    Google Scholar 

  91. Stöber, W., Fink, A. & Bohn, E. Controlled growth of monodisperse silica spheres in the micron size range. J. Colloid Interface Sci. 26, 62–69 (1968).

    Google Scholar 

  92. Severe Acute Respiratory Syndrome Coronavirus 2 Isolate Wuhan-Hu-1, co-Nucleotide-NCBI. https://www.ncbi.nlm.nih.gov/nuccore/MN908947 (2020).

  93. Dai, L. et al. Universal design of Betacoronavirus vaccines against COVID-19, MERS, and SARS. Cell 182, 722–733 (2020).

    Google Scholar 

  94. Stadlbauer, D. et al. SARS-CoV-2 seroconversion in humans: a detailed protocol for a serological assay, antigen production, and test setup. Curr. Protoc. Microbiol. 57, e100 (2020).

    Google Scholar 

  95. Araújo, D. B. et al. SARS-CoV-2 isolation from the first reported patients in Brazil and establishment of a coordinated task network. Mem. Inst. Oswaldo Cruz 115, e200342 (2020).

    Google Scholar 

  96. World Health Organization (WHO). Laboratory Biosafety Guidance Related to the Novel Coronavirus (2019-nCoV). https://www.who.int/docs/default-source/coronaviruse/laboratory-biosafety-novelcoronavirus-version-1-1.pdf?sfvrsn=912a9847_2 (WHO, 2020).

  97. Whittaker, A. L., Liu, Y. & Barker, T. H. Methods used and application of the mouse Grimace Scale in biomedical research 10 years on: a scoping review. Animals 11, 673 (2021).

    Google Scholar 

  98. Schmidt, F. et al. Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses. J. Exp. Med. 217, e20201181 (2020).

    Google Scholar 

  99. Quiros, P. M., Goyal, A., Jha, P. & Auwerx, J. Analysis of mtDNA/nDNA ratio in mice. Curr. Protoc. Mouse Biol. 7, 47–54 (2017).

    Google Scholar 

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