تهیه نانوپارتیکلهای کایتوزان حاوی آنزیم سوپر اکسید دیسموتاز لیشمانیا و بررسی ایمونوژنیسیته آن در موش BALB/c

Abstract

Background Leishmaniasis is a protozoan disease, affecting 12 million people throughout the world. Although several agents are used for treating the disease, the need for long-term therapy, limited efficacy, and development of drug-resistant parasites remain major limitations to effective therapy. MethodsTo develop a new nanovaccine for leishmaniasis, recombinant Leishmania superoxide dismutase B1 (SODB1) was loaded into chitosan nanoparticles by an ionotropic gelation method. The size and loading efficiency of the nanoparticles was evaluated and optimized, and an immunization study was undertaken in BALB/c mice. The mice received phosphate-buffered saline, and SODB1 in phosphate-buffered saline and in nanoparticles, by subcutaneous injection. Soluble Leishmania antigens and complete Freundبs adjuvant were also injected subcutaneously three times at three-weekly intervals (some groups received only a single dose). Three weeks after the last injection, blood samples were collected and assessed using an enzyme-linked immunosorbent assay to detect IgG2a and IgG1.Results Optimized SODB1 loaded nanoparticles had 268 54 nm diameter and Loading efficiency of 71 32%. Immunological study showed that single and triple doses of SODB1 nanoparticles induced significantly higher IgG2a and IgG2a/IgG1 levels than the other groups (P < 0.05).Conclusion Formulations of SODB1 in stable and biodegradable chitosan nanoparticles can increase immunogenicity toward cell-mediated immunity (TH1 cells producing IgG2a in mice) that is effective in Leishmania eradication, and could be developed as a single-dose nanovaccine for leishmaniasis.