Preparation and In Vitro Characterization of Alginate Microspheres Encapsulated with Autoclaved Leishmania major (ALM) and CpG -ODN

Document Type : Original Article

Authors

1 School of Pharmacy, Mashhad University of Medical Sciences, P.O. Box 91775-1365, Mashhad, Iran

2 School of Pharmacy and Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran

3 School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective
The goal of this study was to prepare and characterize alginate microspheres as an antigen delivery
system and adjuvant for immunization against leishmaniasis.
Materials and Methods
Microspheres were prepared by an emulsification technique and characterized for size, encapsulation efficiency, and release profile of encapsulates. Selection of appropriate parameters (viscosity of alginate, emulsifier, and sonication times) enabled the preparation of alginate microspheres with a mean diameter of 1.8 ± 1.0μm, as determined by Scanning Electron Microscopy and Particle Size Analyzer.
Results
The encapsulation efficiency was about 34.2 ± 6.7% for autoclaved leishmania major and 63.5 ± 6.9% for CpG-ODN, as determined by spectrophotometric assays. In vitro release profile showed a slow release rate for encapsulated ALM, while higher release rate was observed for CpG-ODN. The molecular weight was evaluated by SDS-PAGE and showed that the process of encapsulation did not affect the molecular weight of the entrapped antigen.
Conclusion
With regard to the optimum diameter (less than 5 μm), slow release rate and preservation of antigen

Keywords

References

1. Handman E. Leishmaniasis: current status of vaccine development. Clin Microbiol Rev 2001; 14: 229-243.
2. Modabber F. Vaccines against leishmaniasis. Ann Trop Med Parasitol 1995; 89: 83-88.
3. Satti I N, Osman H Y, Daifalla N S, Younis S A, Khalil E A, Zijlstra E E, e t al. Immunogenicity and safety of autoclaved Leishmania major plus BCG vaccine in healthy Sudanese volunteers. Vaccine 2001; 19:2100-2106.
4. Follador I, Araujo C, Orge G, Cheng L H, de Carvalho L P, Bacellar O, et al. Immune responses to an inactive vaccine against American cutaneous leishmaniasis together with granulocyte-macrophage colonystimulating factor. Vaccine 2002; 20: 1365-1368.
5. Fundueanu G, Esposito E, Mihai D, Carpov A, Desbrieres J, Rinaudo M, et al. Preparation and characterization of Ca-alginate microspheres by a new emulsification method. Int J Pharm 1998; 170: 11-21.
6. Lemoine D, Wauters F, Bouchend'homme S, Preat V.Preparation and characterization of alginate microspheres containing a model antigen. Int J Pharm 1998; 176: 9-19.
7. Tafaghodi M, Jaafari M R, Sajadi Tabasi S A. Nasal immunization studies by liposomes encapsulated with tetanus toxoid and CpG-ODN. Eur J Pharm Biopharm 2006; 64: 138-145.
8. Mittal S K, Aggarwal N, Sailaja G, van Olphen A, HogenEsch H, North A, et al. Immunization with DNA, adenovirus or both in biodegradable alginate microspheres: effect of route of inoculation on immune response. Vaccine 2000; 19: 253-263.
9. Gupta R K, Siber G R. Adjuvants for human vaccines--current status, problems and future prospects. Vaccine 1995; 13: 1263-1276.
10. Krieg A M. Mechanisms and applications of immune stimulatory CpG oligodeoxynucleotides.Biochim. Biophys Acta (BBA) - Gene Structure and Express 1999; 1489: 107-116.
11. Mendez S, Tabbara K, Belkaid Y, Bertholet S, Verthelyi D, Klinman D, et al. Coinjection with CpGcontaining immunostimulatory oligodeoxynucleotides reduces the pathogenicity of a live vaccine against cutaneous Leishmaniasis but maintains its potency and durability. Infect Immun 2003; 71: 5121-5129.
12. Rhee E G, Mendez S, Shah J A, Wu C Y, Kirman J R, Turon T N, et al. Vaccination with heat-killed leishmania antigen or recombinant leishmanial protein and CpG oligodeoxynucleotides induces long-term memory CD4+ and CD8+ T cell responses and protection against leishmania major infection. J Exp Med 2002; 195: 1565-1573.
13. Verthelyi D, Kenney R T, Seder R A, Gam A A, Friedag B, Klinman D M. CpG oligodeoxynucleotides as vaccine adjuvants in primates. J Immunol 2002; 168: 1659-1663.
14. Diwan M, Tafaghodi MSamuel J. Enhancement of immune responses by co-delivery of a CpG oligodeoxynucleotide and tetanus toxoid in biodegradable nanospheres. J Control Rel 2002; 85: 247-262.
15. Tafaghodi M. Nasal immunization using biodegradable microspheres and liposomes: tetanus toxoid as a model. Ph.D. Thesis, Mashhad: University of Medical Sciences; 2003.
16. Tafaghodi M, Sajadi Tabasi S A, Jaafari M R. Induction of systemic and mucosal immune responses by intranasal administration of alginate microspheres encapsulated with tetanus toxoid and CpG-ODN. Int J Pharm 2006; 319: 37-43.
17. Tafaghodi M, Sajadi Tabasi S A, Jaafari M R. Formulation and characterization and release studies of alginate microsphere encapsulated with tetanus toxoid. J Biomat Sci Polymer Ed 2006; 17: 909-924.
18. Waterborg J H. Quantitation of proteins. In: Walker J. (ed.). The Protein Protocols Handbook. Newjersey: Humana Press; 2002. 3-36. ALM microspheres
19. Barman S P, Lunsford L, Chambers P, Hedley M L. Two methods for quantifying DNA extracted from poly (lactide-co-glycolide) microspheres. J Control Rel 2000; 69: 337-344.
20. Sinha V R, Trehan A. Biodegradable microspheres for protein delivery. J Control Rel 2003; 90: 261-280.
21. Vandenberg G W, Drolet C, Scott S L, De la Noue J. Factors affecting protein release from alginatechitosan coacervate microcapsules during production and gastric/intestinal simulation. J Control Rel 2001; 77: 297-307.
22. Al-Musa S, Abu Fara D, Badwan A A. Evaluation of parameters involved in preparation and release of drug loaded in crosslinked matrices of alginate. J Control Rel 1999; 57: 223-232.
23. Freiberg S, Zhu X X. Polymer microspheres for controlled drug release. Int J Pharm 2004; 282: 1-18.
24. Freitas S, Merkle H P, Gander B. Microencapsulation by solvent extraction/evaporation: reviewing the state of the art of microsphere preparation process technology. J Control Rel 2005; 102: 313-332.
25. Kahl L P, Lelchuk R, Scott C A, Beesley J. Characterization of Leishmania major antigen-liposomes that protect BALB/c mice against cutaneous leishmaniasis. Infect Immun 1990; 58: 3233-3241.

Files

Share

How to cite

Statistics