Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

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
Download PDF
⟨ Previous Next ⟩

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.

References

  1. Yang Y, Ronge X, Song L, Yukun Q, Kecheng L, Huahua Y, et al. 2020. Chitosan, hydroxypropyltrimethyl ammonium chloride chitosan and sulfated chitosan nanoparticles as adjuvants for inactivated newcastle disease vaccine. Carbohydr. Polym. 229: 115423. 
  2. Carroll EC, Jin L, Mori A, Munoz-Wolf N, Oleszycka E, Moran HBT, et al. 2016. The vaccine adjuvant chitosan promotes cellular immunity via DNA sensor cGAS-STING-dependent induction of type I interferons. Immunity 44: 597-608.  https://doi.org/10.1016/j.immuni.2016.02.004
  3. El-Sissi AF, Farida HM, Nadia MD, Ali QG, Korany AA. 2020. Chitosan and chitosan nanoparticles as adjuvant in local rift valley fever inactivated vaccine. 3 Biotech. 10: 88. 
  4. Biswas, S, Chattopadhyay M, Sen KK, Saha, MK. 2015. Development and characterization of alginate coated low molecular weight chitosan nanoparticles as new carriers for oral vaccine delivery in mice. Carbohydr. Polym. 121: 403-410.  https://doi.org/10.1016/j.carbpol.2014.12.044
  5. Bachmann MF, Jennings GT. 2010. Vaccine delivery: a matter of size, geometry, kinetics and molecular patterns. Nat. Rev. Immunol. 10: 787-796.  https://doi.org/10.1038/nri2868
  6. Manolova V, Flace A, Bauer M, Schwarz K, Saudan P, Bachmann MF. 2008. Nanoparticles target distinct dendritic cell populations according to their size. Eur. J. Immunol. 38: 1404-1413.  https://doi.org/10.1002/eji.200737984
  7. Bootz A, Karbach A, Spindler J, Kropff B, Reuter N, Sticht H, et al. 2017. Protective capacity of neutralizing and non-neutralizing antibodies against glycoprotein B of cytomegalovirus. PLoS Pathog. 13: e1006601. 
  8. Wang H, Huang C, Dong J, Yao Y, Xie Z, Liu X, et al. 2015. Complete protection of mice against lethal murine cytomegalovirus challenge by immunization with DNA vaccines encoding envelope glycoprotein complex III antigens GH, GL and GO. PLoS One 10: e0119964. 
  9. Li L, Freed DC, Liu Y, Li F, Barrett DF, Xiong W, et al. 2021. A Conditionally replication-defective cytomegalovirus vaccine elicits potent and diverse functional monoclonal antibodies in a phase I clinical trial. Npj Vaccines 6: 79. 
  10. Bialas KM, Tanaka T, Tran D, Varner V, Cisneros De La Rosa E, Chiuppesi F, et al. 2015. Maternal CD4+ T cells protect against severe congenital cytomegalovirus disease in a novel nonhuman primate model of placental cytomegalovirus transmission. Proc. Natl. Acad. Sci. USA 112: 13645-13650.  https://doi.org/10.1073/pnas.1511526112
  11. Bongard N, Le-Trilling VTK, Malyshkina A, Ruckborn M, Wohlgemuth K, Wensing I, et al. 2019. Immunization with a murine cytomegalovirus based vector encoding retrovirus envelope confers strong protection from friend retrovirus challenge infection. PLoS Pathog. 15: e1008043. 
  12. Almalik A, Donno R, Cadman CJ, Cellesi F, Day PJ, Tirelli N. 2013. Hyaluronic acid-coated chitosan nanoparticles: molecular weight-dependent effects on morphology and hyaluronic acid presentation. J. Control. Release 172: 1142-1150.  https://doi.org/10.1016/j.jconrel.2013.09.032
  13. Jesus S, Borchard G, Borges O. 2013. Freeze dried chitosan/ poly-ε-caprolactone and poly-ε-caprolactone nanoparticles: evaluation of their potential as DNA and antigen delivery systems. J. Genet. Syndr. Gene Ther. 4: 7. 
  14. Bento D, Jesus S, Lebre F, Goncalves T, Borges O. 2019. Chitosan plus compound 48/80: formulation and preliminary evaluation as a hepatitis B vaccine adjuvant. Pharmaceutics 11: 72. 
  15. Kilkenny C, Browne WJ, Cuthi I, Emerson M, Altman DG. 2012. Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. Vet. Clin. Pathol. 41: 27-31.  https://doi.org/10.1111/j.1939-165X.2012.00418.x
  16. Mauricio-Sanchez RA, Salazar R, Luna-Barcenas JG, Mendoza-Galvan A. 2018. FTIR spectroscopy studies on the spontaneous neutralization of chitosan acetate films by moisture conditioning. Vib. Spectrosc. 94: 1-6.  https://doi.org/10.1016/j.vibspec.2017.10.005
  17. Slovakova M, Kratochvilova V, Palarcik J, Metelka R, Dvorakova P, Srbova J, et al. 2016. Chitosan nanofibers and nanoparticles for immobilization of microbial collagenase. Fibers Textile 23: 193-198. 
  18. de la Calle I, Soto-Gomez D, Perez-Rodriguez P, Lopez-Periago JE, 2019. Particle size characterization of sepia ink eumelanin biopolymers by SEM, DLS, and AF4-MALLS: a comparative study. Food Anal. Methods 12: 1140-1151.  https://doi.org/10.1007/s12161-019-01448-0
  19. Correia-Pinto J, Csaba N, Schiller J, Alonso M. 2015. Chitosan-poly (I:C)-PADRE based nanoparticles as delivery vehicles for synthetic peptide vaccines. Vaccines 3: 730-750.  https://doi.org/10.3390/vaccines3030730
  20. Li Z, Li Y, Wang Y, Hou Y, Cao H, Wu X, et al. 2021. Intranasal immunization with a RNMB0315 and combination adjuvants induces protective immunity against Neisseria meningitidis serogroup B in mice. Int. Immunopharmacol. 93: 107411. 
  21. Borges O, Cordeiro-da-Silva A, Tavares J, Santarem N, de Sousa A, Borchard G, et al. 2008. Immune response by nasal delivery of hepatitis B surface antigen and codelivery of a CpG ODN in alginate coated chitosan nanoparticles. Eur. J. Pharm. Biopharm. 69: 405-416.  https://doi.org/10.1016/j.ejpb.2008.01.019
  22. Wen ZS, Xu YL, Zou XT, Xu ZR. 2011. Chitosan nanoparticles act as an adjuvant to promote both Th1 and Th2 immune responses induced by ovalbumin in mice. Mar. Drugs 9: 1038-1055. https://doi.org/10.3390/md9061038
  23. Li X, Xing R, Xu C, Liu S, Qin Y, Li K, et al. 2021. Immunostimulatory effect of chitosan and quaternary chitosan: a review of potential vaccine adjuvants. Carbohydr. Polym. 264: 118050. 
  24. Rampino A, Borgogna M, Blasi P, Bellich B, Cesaro A. 2013. Chitosan nanoparticles: preparation, size evolution and stability. Int. J. Pharm. 455: 219-228.  https://doi.org/10.1016/j.ijpharm.2013.07.034
  25. Dong C, Wang Y, Gonzalez GX, Ma Y, Song Y, Wang S, et al. 2021. Intranasal vaccination with influenza HA/GO-PEI nanoparticles provides immune protection against homo- and heterologous strains. Proc. Natl. Acad. Sci. USA 118: e2024998118. 
  26. Nasti A, Zaki NM, de Leonardis P, Ungphaiboon S, Sansongsak P, Rimoli MG, et al. 2009. Chitosan/TPP and chitosan/TPP-hyaluronic acid nanoparticles: systematic optimisation of the preparative process and preliminary biological evaluation. Pharm. Res. 26: 1918-1930.  https://doi.org/10.1007/s11095-009-9908-0
  27. Wang Y, Li P, Kong L. 2013. Chitosan-modified PLGA nanoparticles with versatile surface for improved drug delivery. AAPS PharmSciTech. 14: 585-592.  https://doi.org/10.1208/s12249-013-9943-3
  28. da Silva Furtado GTF, Fideles TB, de Cassia Alves Leal Cruz R, de Lima Souza JW, Barbero MAR, Fook MVL. 2020. Chitosan/NaF particles prepared via ionotropic gelation: evaluation of particles size and morphology. Mat. Res. 21. doi.org/10.1590/1980-5373-MR-2018-0101 . 
  29. Azevedo JR, Sizilio RH, Brito MB, Costa AMB, Serafini MR, Araujo AAS, et al. 2011. Physical and chemical characterization insulin-loaded chitosan-TPP nanoparticles. J. Therm. Anal. Calorim. 106: 685-689.  https://doi.org/10.1007/s10973-011-1429-5
  30. Jin Z, Gao S, Cui X, Sun D, Zhao K. 2019. Adjuvants and delivery systems based on polymeric nanoparticles for mucosal vaccines. Int. J. Pharm. 572: 118731. 
  31. Foster S, Duvall CL, Crownover EF, Hoffman AS, Stayton PS. 2010. Intracellular delivery of a protein antigen with an endosomal-releasing polymer enhances CD8 T-cell production and prophylactic vaccine efficacy. Bioconjug. Chem. 21: 2205-2212.  https://doi.org/10.1021/bc100204m
  32. Farace C, Sanchez-Moreno P, Orecchioni M, Manetti R, Sgarrella F, Asara Y, et al. 2016. Immune cell impact of three differently coated lipid nanocapsules: pluronic, chitosan and polyethylene glycol. Sci. Rep. 6: 18423. 
  33. Borges O, Tavares J, de Sousa A, Borchard G, Junginger HE, Cordeiro-da-Silva A. 2007. Evaluation of the immune response following a short oral vaccination schedule with hepatitis B antigen encapsulated into alginate-coated chitosan nanoparticles. Eur. J. Pharm. Sci. 32: 278-290. https://doi.org/10.1016/j.ejps.2007.08.005