Antimalarial efficacy of low molecular weight chitosan against Plasmodium berghei infection in mice.

BACKGROUND & OBJECTIVES: Despite continuous global attempts to fight parasitic infections, malaria still remains one of the major human life threatening diseases. Difficulty of producing efficient antimalaria vaccines and increasing drug-resistant strains, highlight the urgent need to search for a new alternative antimalaria drug. The aim of this study was to find a new agent against malaria parasite with maximum efficacy and minimum range of side-effects. For this, the antiplasmodial activity of commercial chitosan, a natural carbohydrate polymer, was evaluated on Plasmodium berghei via in vivo experiments. This is the first report that to highlight antimalarial effects of low molecular weight chitosan against P. berghei in vivo. METHODS: Low molecular weight chitosan with 95% degree of deacetylation was melted in normal saline with 1% (w/v) acetic acid for preparing 10, 20, 40 and 80 mg/kg concentrations of chitosan, which were then examined for their antimalarial efficacy in P. berghei infected mice. RESULTS: The study showed that differrent concentrations of chitosan exhibited significant antimalarial effect (p= 0.002) when compared with the control group. Also, analysis of mice survival time showed significant differences between 20 and 80 mg/kg concentrations of used chitosan in comparison to negative control group. INTERPRETATION & CONCLUSION: The results of this study showed that the chitosan has potent antimalarial activity and could be suggested as an alternative antimalarial drug component.

Assessment of the immunogenicity and protective efficiency of a novel dual-promoter DNA vaccine, harboring SAG1 and GRA7 genes, from RH strain of Toxoplasma gondii in BALB/c mice.

BACKGROUND: Toxoplasmosis, a protozoan parasitic disease caused by Toxoplasma gondii, has been a serious human and veterinary medicine problem with global distribution. In the current study, we assessed immunogenicity and protective efficiency of a novel dual-promoter DNA vaccine, harboring SAG1 and GRA7 genes, from RH strain of Toxoplasma gondii (T. gondii) with or without CpG-ODN as adjuvant in a murine model. METHODS: BALB/c mice were immunized intramuscularly with pVitro-SAG1-GRA7 alone and pVitro-SAG1-GRA7 with CpG-ODN three times at three-week intervals. Enzyme-linked immunosorbent assay (ELISA) was used to assess total IgG, IgG1 and IgG2a antibodies and gamma interferon (IFN-γ) and interleukin-10 (IL-10) cytokines in mice sera. Four weeks post final vaccination, MTT assay and lethal challenge-infection with 1×103 tachyzoites of T. gondii RH strain were carried out to assess stimulation index (SI) and mice survival time, respectively. RESULTS: The IgG levels in mice immunized with multicomponent vaccines, including pVitro-SAG1-GRA7 alone and pVitro-SAG1-GRA7 with CpG-ODN, were significantly higher than those in control mice or single-gene DNA-vaccinated ones (P<0.05). Furthermore, level of IgG2a in mice receiving pVitro-SAG1-GRA7 with CpG-ODN was significantly higher than that in mice receiving pVitro-SAG1-GRA7 alone (P<0.05). The Toxoplasma lysate antigen (TLA)-stimulated lymphocytes from pVitro-SAG1-GRA7 with CpG-ODN group responded more dramatically than those from control groups or single-gene DNA-vaccinated groups (P<0.001). The pVitro-SAG1-GRA7 with CpG-ODN-vaccinated mice developed high levels of IgG2a and IFN-γ (P<0.001) and low levels of IgG1 and IL-10, compared to control groups, suggesting a modulated immune response type Th1. In addition, survival time of the mice immunized with pVitro-SAG1-GRA7 with CpG-ODN was significantly extended, compared to controls (P<0.05); however, all mice died. CONCLUSION: The multivalent pVitro-SAG1-GRA7 DNA vaccine with CpG-ODN adjuvant is a promising vaccine candidate against toxoplasmosis.

Anti-Toxoplasma activity of various molecular weights and concentrations of chitosan nanoparticles on tachyzoites of RH strain.

BACKGROUND: Natural polysaccharides such as chitosan (CS) are widely used as antimicrobial agents. In recent years, and considering that CS has a strong antimicrobial potential, interest has been focused on antimicrobial activity of chitosan nanoparticles (CS NPs). The main factors affecting the antibacterial activity of chitosan include molecular weight (MW) and concentration. In this regard, the aim of this study was to produce various MWs and concentrations of CS NPs, through the ionic gelation method, and investigate their potential anti-parasitic activity against tachyzoites of Toxoplasma gondii RH strain. MATERIALS AND METHODS: The MWs and degree of deacetylation of the CS were characterized using viscometric and acid-base titration methods, respectively. The efficacy of various MWs and concentrations of NPs was assessed by performing in vitro experiments for tachyzoites of T. gondii RH strain, such as MTT assay, scanning electron microscopy, bioassay in mice and PCR. In vivo experiment was carried out in BALB/c mice which were inoculated with tachyzoites of T. gondii RH strain and treated with various MWs of CS NPs. RESULTS: The results of in vitro and in vivo experiments revealed that anti-Toxoplasma activity strengthened as the CS NPs concentration increased and the MW decreased. In vitro experiment showed 100% mortality of tachyzoites at 500 and 1,000 ppm concentrations of low molecular weight (LMW) CS NPs after 180 min and at 2,000 ppm after 120 min. Furthermore, a 100% mortality of tachyzoites was observed at 1,000 and 2,000 ppm concentrations of medium molecular weight (MMW) CS NPs and at 2,000 ppm concentration of high molecular weight (HMW) CS NPs after 180 min. Growth inhibition rates of tachyzoites in peritoneal exudates of mice receiving low, medium and high MWs of CS NPs were found to be 86%, 84% and 79% respectively, compared to those of mice in sulfadiazine treatment group (positive control). CONCLUSION: Various MWs of CS NPs exhibited great anti-Toxoplasma efficiency against tachyzoites of RH strain, with the greatest efficacy shown by LMW CS NPs in both experiments. It seems that CS NPs can be used as an alternative natural medicine in the treatment of toxoplasmosis.