Preparation and Evaluation of Immunogenicity of Chitosan Nanoparticles Containing Tetanotoxin in Comparison with Freund\'s Adjuvant in Mouse Model for Improvement of Tetanus Vaccine after Subcutaneous Injection

Abstract

Background and Objectives: Tetanus is a deadly zoonotic infectious disease. For nearly a century, tetanus vaccine has been produced and used. This study was carried out with the aim of preparation and evaluation of immunogenicity of chitosan nanoparticles containing tetanotoxin in comparison with Freund's adjuvant in mouse model for improvement of tetanus vaccine after subcutaneous onjection. Methods: In this experimental study, chitosan nanoparticles synthesis containing tetanus toxoid was performed using ionic gelation method. The morphology of nanoparticles was investigated using SEM and AFM, then, the nanoparticles were subcutaneously injected into mice and two weeks after the end of injection, blood was taken. Antibody titer was measured by indirect ELISA. Data were analyzed using one-way one-way ANOVA and Tukey test at significance level of p<0.05. Results: High-load antigen (uniform spherical morphology and the mean size of 144), were encapsulated in chitosan nanoparticles. After serum analysis, IgG antibody titer was significantly different between the mice that received antigen as nanoparticle and the control group, but no significant difference was observed in the antigen with Freund’s adjuvant group. Conclusion: Based on the results of this study, tetanus toxoid injection using nanoproces is able to induce humoral immune response almost equal to Freund’s adjuvant and can be an appropriate alternative.

Keywords

• nanoparticles
• tetanus
• chitosan
• freund’s adjuvant