A candidate vaccine composed of live nonpathogenic Iranian Lizard Leishmania mixed with Chitin microparticles protects mice against Leishmania major infection.

BACKGROUND: The protective effect of immunization using Iranian Lizard Leishmania (ILL) mixed with CpG oligodeoxynucleotides (CpG-ODN) was demonstrated in a previous study. Here, we report the effect of leishmanization using ILL mixed with chitin microparticles (CMPs) as an adjuvant against L. major infection in BALB/c mice. METHODS: Briefly, 2 × 107 live ILL were mixed with 10 µg CMPs (<40 µm in size) (ILL+CMP) and were injected subcutaneously into the right footpad of BALB/c mice. Three control groups were included in the study and received ILL, chitin, and PBS respectively. Three weeks later, mice were challenged with 2 × 105 live L. majorEGFP promastigotes, which were inoculated into the left footpad. The infection course was monitored using footpad swelling measurement and in vivo imaging. Eleven weeks after the challenge, all mice were sacrificed and parasite burden was measured in the spleen and the draining lymph node using three different methods including real-time PCR, flow cytometry, and direct fluorescent microscopy. In addition, cytokines levels (IFN-γ and IL-10), and nitric oxide production were assayed in splenocytes. RESULTS: Mice immunized with ILL+CMP had a smaller footpad diameter in comparison to control groups and notably, no lesion was developed at the inoculation site. Additionally, in vivo imaging study revealed that there was no detectable fluorescence in the ILL+CMP group footpad by the end of the tenth week. This finding was confirmed by three methods used for parasite burden assays. Moreover, higher IFN-γ level was observed in mice immunized with ILL+CMP in comparison with other groups. On the other hand, nitric oxide concentration was higher in the ILL control group. CONCLUSION: ILL mixed with chitin microparticles is an effective vaccine against leishmaniasis in BALB/c mice. This vaccine is able to induce an adequate immune response to decrease the parasite burden and prevent lesion formation. Further studies are needed to evaluate long-lasting immunity, especially in experimental outbreed models.

Oral chitin treatment improved demyelination in murine autoimmune encephalomyelitis model by inhibition of inflammatory responses.

This study aimed to determine whether chitin microparticles (CMP), glucosamine-based polymers, have an anti-inflammatory response in a murine model of autoimmune encephalomyelitis. Experimental autoimmune encephalomyelitis (EAE) was induced in C57BL/6 mice by immunization with myelin antigens emulsified in complete Freund adjuvant. A standard clinical and histological method (Luxol Fast Blue staining) was used to validate the model and document the impact of CMP treatment. ELISA was used to determine the production of spleen cell cytokines and serum levels of anti-chitin antibodies. Flowcytometry was used to determine the percentage of regulatory lymphocytes. The relative expression of the breast regression protein 39 (BRP-39) gene was examined through real time-PCR amplification. Clinical signs were significantly improved in mice given CMP compared with untreated mice. Histological analysis of the spinal cord revealed that treatment significantly reduced demyelination. The levels of interferon-γ, interleukin-17, and tumor necrosis factor-α were also reduced; conversely, no significant change was detected in interleukin-10 level and regulatory T cell count. The CMP-fed mice showed lower BRP-39 expression compared with the control group. It was ultimately determined that CMP modulates immune responses which could indirectly alter the pathology of an injured central nervous system. The data suggests that CMP may be used as an effective and cheap oral therapeutic agent for multiple sclerosis.