Effect of interleukin-22 on immunogenicity of DNA vaccine encoding TSA gene of leishmania major in BALB/c mice

Background and purpose: Previous Research shows the use of plasmids containing genes TSA to be useful as vaccines for Leishmania major. Recently, the role of interleukin-22 (IL-22) in tissue repair has been demonstrated. In this research, the effect of IL-22 on encoding TSA gene of Leishmania major in BALB/c mice was assessed

Expression of complete rhoptry protein 2 (ROP2) gene of Toxoplasma gondii in eukaryotic cell

Toxoplasma gondii is the intracellular protozoan parasite responsible for animal and human toxoplasmosis. In immunodeficient patients, chronic infection with T. gondii can reactivate and produceencephalitis, which is often lethal. ROP2 (rhoptry protein of T. gondii) is one of the most important interferer in organelle and PVM blending. ROP2 protein is recognized by clone T-cell (Tcc32) in humanbody and also has epitope for B-cell. All of these characteristics of ROP2 makes it a candidate for cocktail vaccine and recombinant vaccine against toxoplasmosis. We described the expression of thegene which encodes the complete rhoptry protein 2 (ROP2) of T. gondii in CHO cells and confirmed it by SDS-PAGE and Western blot analysis. In the present work, genomic DNA of T. gondii was extractedand used for amplifying of ROP2 gene as a template. Then PCR product was cloned into pTZ57R/T vector, and plasmid containing ROP2 gene (pT-ROP2) was extracted from transformed bacteria andsequenced. We hope to use from this recombinant plasmid (pT-ROP2) to make DNA vaccine against toxoplasmosis

Effect of IL-22 on DNA vaccine encoding LACK gene of Leishmania major in BALB/c mice

In the present study, the effect of IL-22 together with the plasmid encoding LACK (Leishmania homolog of receptors for activated C-kinase) gene of Leishmania major on the trend of leishmaniasis in BALB/c mice was evaluated.Evaluation of the cellular and humoral immunity was performed by measurement of IL-4 and IFN-γ, culture of splenocytes and MTT assay, and measurement of total IgG, IgG1, and IgG2a in the control and immunized groups. Clinical evaluations were also carried out by measurement of the lesion size, survival rate, and body weight of mice.Comparison of the mean size of lesions in the LACK and LACK. +. IL-22 groups demonstrated that the mean size of lesions of the two groups was significantly different from week four (p<. 0.05).The survival rate at day 170 after challenge for the PBS, pcDNA3 (empty plasmid), pcLACK (pcDNA3 containing LACK gene), and pcLACK. +. IL-22 groups were 20%, 40%, 60%, and 80%, respectively.According to the results of IFN-γ, IL-4, total IgG, IgG1, and IgG2a measurement and the MTT assay, IL-22 obviously caused an increase in IFN-γ production and a decrease in IL-4 production before and after the challenge (p<. 0.05). The results showed the effectiveness of IL-22 in DNA vaccine. It showed that IL-22 brought about Th1 cytokine responses and high survival rate of mice. © 2013 Elsevier Inc

DNA vaccination with a plasmid encoding LACK-TSA fusion against Leishmania major infection in BALB/c mice

Vaccination would be the most important strategy for the prevention and elimination of leishmaniasis. The aim of the present study was to compare the immune responses induced following DNA vaccination with LACK (Leishmania analogue of the receptor kinase C), TSA (Thiol-specific-antioxidant) genes alone or LACK-TSA fusion against cutaneous leishmaniasis (CL). Cellular and humoral immune responses were evaluated before and after challenge with Leishmania major (L. major). In addition, the mean lesion size was also measured from 3th week post-infection. All immunized mice showed a partial immunity characterized by higher interferon (IFN)-gamma and Immunoglobulin G (IgG2a) levels compared to control groups (p< 0.05). IFN-gamma/Interleukin (IL)-4 and IgG2a/IgG1 ratios demonstrated the highest IFN-gamma and IgG2a levels in the group receiving LACK-TSA fusion. Mean lesion sizes reduced significantly in all immunized mice compared with control groups at 7th week post-infection (p< 0.05). In addition, there was a significant reduction in mean lesion size of LACK-TSA and TSA groups than LACK group after challenge (p< 0.05). In the present study, DNA immunization promoted Th1 immune response and confirmed the previous observations on immunogenicity of LACK and TSA antigens against CL. Furthermore, this study demonstrated that a bivalent vaccine can induce stronger immune responses and protection against infectious challenge with L. major

Partial immunity in murine by immunization with a toxoplasmic DNA vaccine

Toxoplasma gondii is an obligate intracellular protozoan that is causative agent of atoxoplasmosis, a disease which may result in a spectrum of consequences. Previous studies have reported that DNA vaccine can be effective in partial protection against this parasite. In this study, we constructed a single DNA vaccine containing rhoptry protein 1 (ROP1) and evaluated its immune response in Balb/c mice. We used alum as an adjuvant to enhance the immune response. After intramuscular injection, we evaluated the immune response using cytokine and antibody assay and mortality rate. The results show that mice immunized by pcROP1 with or without alum produced high Th1 immune response compared with the control groups. This kind of DNA vaccine prolonged survival time. The current study showed that ROP1 DNA vaccine could induce partial protective response against toxoplasmosis.Key words: Toxoplasma gondii, DNA, vaccine, Rhoptry protein 1 (ROP1)

A systematic review of the effects of Iranian pharmaceutical plant extracts on Giardia lamblia

This study aimed to provide a systematic review regarding anti-Giardia effect of different Iranian plant extracts used in vivo and in vitro on cysts and trophozoites. Many reports indicated that most of plant extracts used as anti-Giardia were obtained from Liliaceae, Apiaceae, Asteraceae, and Myrtaceae. These extracts included different fractions such as aqueous, alcoholic and chloroform extracts as well as Soxhlet extraction of juice or essence. The findings of this review showed that hydroalcoholic extract of asafoetida, Chenopodium botrys, and chloroformic extract of feverfew (Tanacetum parthenium) have the maximum effect (100 efficacy) on in vitro application against Giardia. However, the highest in vivo effect of 100 therapeutic significance was recorded for the extract of Allium sativum at 80 mg/mL concentration. Given the plant species richness of Iran in terms of herbal medicines with fewer side effects, it can be a good alternative to chemical drugs used to treat giardiasis. © 2015 Asian Pacific Tropical Medicine Press

The Effect of Vitamin D3 Alone and Mixed With IFN-γ on Tachyzoites of Toxoplasma gondii (RH Strain) Proliferation and Nitric Oxide (NO) Production in Infected Macrophages of BALB/C Mice

Introduction: Toxoplasma gondii is an obligatory interacelullar parasite that infects nucleated cells in its intermediate hosts. The aim of the present study was to determine the effect of vitamin D3 on the multiplication of T. gondii in peritoneal macrophage of Balb/c mice and nitric oxide production by macrophages. Methods: According to usage of vitamin D3 (one dose or seven doses) and INFγ in vitro and in vivo, this study was divided into four experiments. In all experiments, the macrophages were col­lected from peritoneum and cultured in RPMI-1640. Then the supernatants were collected after 24 h and their nitric oxide was measure. After 96 h, the macrophages were collected and stained and the number of tachyzoites was measured. Results: The first experiment (the mice were infected with tachyzoites and after 2 h, got one dose vita­min D3 intraperitonealy) showed the best results. The mean of tachyzoites per macrophages was 2.37, and mean ± SD of nitric oxide was 187.8 ± 9.Discussion: High-level production of nitric oxide may be related to the only one injection of vita­min D3. The injection in long time might suppress the immune system

Molecular assessment of Neospora caninum and Toxoplasma gondii in hooded crows (Corvus cornix) in Tehran, Iran

Abstract Neospora caninum and Toxoplasma gondii are two closely related protozoan parasites that have been detected from various species of bird hosts. However, little is known about the prevalence of N. caninum and T. gondii in crows. Hence, we examined the molecular frequency of N. caninum and T. gondii in the brain samples of hooded crows (Corvus cornix) that collected from different public parks of Tehran, Iran by nested-PCR method. We used the primers targeting the Nc5 and GRA6 genes for detection of N. caninum and T. gondii, respectively. From a total of 55 brain samples, 5 (9.9%) and 9 (16.36%) samples were positive for N. caninum and T. gondii, respectively. Sequencing of a N. caninum isolate revealed 95%–100% identity with the deposited N. caninum in GenBank. Genotyping of T. gondii isolates by PCR-RFLP analysis of the GRA6 gene revealed type III genotype in 8 isolates. The results of this study indicate that hooded crows may have a putative role in transmission of N. caninum and T. gondii to canines and felines definitive hosts, respectively