Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Nasal Immunization Using Chitosan Nanoparticles with Glycoprotein B of Murine Cytomegalovirus

  • Marcela Slovakova (University of Defence, Faculty of Military Health Sciences) ;
  • Sylva Janovska (University of Defence, Faculty of Military Health Sciences) ;
  • Radek Sleha (University of Defence, Faculty of Military Health Sciences) ;
  • Vera Radochova (University of Defence, Faculty of Military Health Sciences) ;
  • Alexandra Hatala (University of Pardubice, Faculty of Chemical Technology) ;
  • Nikola Mannova (University of Pardubice, Faculty of Chemical Technology) ;
  • Radovan Metelka (University of Pardubice, Faculty of Chemical Technology) ;
  • Ludovit Pudelka (University of Defence, Faculty of Military Health Sciences) ;
  • Pavel Bostik (University of Defence, Faculty of Military Health Sciences)
  • Received : 2023.08.08
  • Accepted : 2023.12.14
  • Published : 2024.03.28
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Abstract

The use of nanoparticles as a delivery system for a specific antigen could solve many limitations of mucosal vaccine applications, such as low immunogenicity, or antigen protection and stabilization. In this study, we tested the ability of nasally administered chitosan nanoparticles loaded with glycoprotein B of murine cytomegalovirus to induce an immune response in an animal model. The choice of chitosan nanoparticle type was made by in vitro evaluation of sorption efficiency and antigen release. Three types of chitosan nanoparticles were prepared: crosslinked with tripolyphosphate, coated with hyaluronic acid, and in complex with polycaprolactone. The hydrodynamic size of the nanoparticles by dynamic light scattering, zeta potential, Fourier transform infrared spectroscopy, scanning electron microscopy, stability, loading efficiency, and release kinetics with ovalbumin were evaluated. Balb/c mice were immunized intranasally using the three-dose protocol with nanoparticles, gB, and adjuvants Poly(I:C) and CpG ODN. Subsequently, the humoral and cell-mediated antigen-specific immune response was determined. On the basis of the properties of the tested nanoparticles, the cross-linked nanoparticles were considered optimal for further investigation. The results show that nanoparticles with Poly(I:C) and with gB alone raised IgG antibody levels above the negative control. In the case of mucosal IgA, only gB alone weakly induced the production of IgA antibodies compared to saline-immunized mice. The number of activated cells increased slightly in mice immunized with nanoparticles and gB compared to those immunized with gB alone or to negative control. The results demonstrated that chitosan nanoparticles could have potential in the development of mucosal vaccines.

Keywords

Acknowledgement

This research was supported by the Ministry of Education Youth and Sports (the program InterExcellence-InterAction, grant no. LTAUSA-19003), the Ministry of Defence of the Czech Republic (long-term organization development plan) and by the Ministry of Health (MH-CZ DRO (UHHK, 00179906). The authors thank Ing. Jan Bartacek, Ph.D. for the FTIR analysis, and doc. Ing. Libor Cervenka, Ph.D. and doc. Ing. Libor Cervenka, Ph.D. for lyophilisation. The authors are grateful to Ian McColl MD. Ph.D for language assistance with the manuscript. The authors are grateful to Ian McColl MD PhD for language assistance with the manuscript.

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