Brucellosis is a zoonotic disease in human and animals. Brucella melitensis is one of the most pathogenic species of Brucella in goat and sheep. Omp31 is an outer membrane protein of Brucella that acts as an immunogenic protein. Cytokines are glycoproteins with low molecular weight that play the role of an immune adjuvant and regulate immune responses. Interleukin-2 is one of the most important cytokines, which are secreted by the white blood cells and involved in T cell immune responses. In the present study, a chimeric Omp31-Interleukin2 recombinant protein was generated by means of genetic engineering techniques. This chimeric coding sequence was amplified by using specific primers and using Splicing Overlap Extension (SOE) PCR technique. The fusion of the two mentioned proteins was accomplished using a rigid linker. The generated chimeric IL2-Omp31 fragment was TA cloned, and then subcloned into pEt22b vector as an expression vector. The chimeric protein was successfully expressed in E. coli BL21 (DE3) and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and also Western-blotting analysis. Finally, in order to assess the antigenic features of the recombinant chimeric IL2-Opm31 protein, its secondary structure and antigenicity were predicted in silico.

Background: Denileukin diftitox is a recombinant immunotoxin composed of truncated diphtheria toxin fused to human interleukin 2 (DT389-IL2). It is a candidate for protein therapy of lymphoma. This work aims to investigate DT389-IL2 production using a recombinant strain of Escherichia coli in the lab-scale bioreactor. Materials and Methods: First, the effect of chemical composition in culture medium (the complex and defined) was investigated in DT389-IL2 expression in E. coli BL21 (DE3) containing pET-IDZ plasmid in shack flask and lab-bioreactor culture. To enrich the carbon source, we added glucose 6-8 g/L to the complex culture medium. The composition of the defined medium was enriched by adding amino acids (valine, 0. 0502; phenylalanine, 0. 0132; lysine, 0. 0184; aspartic acid, 0. 016; and serine, 0. 0251 g/L). The growth cell was measured by the optical density method and the expression yield was investigated by SDS-PAGE. Results: The maximum growth rate was determined in a modified complex culture medium containing glucose (6 g/L). The bacteria growth in the medium was achieved to the optical density of 5. 5 in the shack flask culture, and under the same condition, the optical density of medium increased to 13 in the bioreactor. Adding amino acids to the defined medium significantly enhanced cell growth and expression of proteins. Also, the optical densities of 2. 95 and 6. 1 were achieved in the shack-flask and bioreactor culture, respectively. The specific growth rates of bacteria in the complex medium and the defined medium were determined as 0. 89 and 0. 47 h-1, respectively. Conclusion: Adding glucose (carbon source) to the complex medium and elemental enrichment by adding amino acids increased biomass production. The growth cell rate in the bioreactor through suitable control of the environmental conditions and proper mixing was higher compared with the shack-flask culture.

Objective: Multiple sclerosis (MS) is a chronic autoimmune disease due to demyelination of the central nervous system. It is believed that cytokines are involved in the pathogenesis of MS. The interleukin-2 (IL2) gene is powerful functional candidate that is involved in immune regulation and operation. In this study, for the first time, we investigated the effect of -475 A/T and -631 G/A IL2 polymorphisms on MS disease in Iranian patients.Materials and Methods: In this case-control study, 100 MS patients (mean age: 32.95±6.51 years, age range: 20-42 years) selected according to McDonald criteria, and 100 ethnically, sex and age matched healthy controls (mean age: 29±7.8 years, age range: 20-52 years) with no personal or family history of autoimmune diseases were studied. The restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method was applied to define different alleles and genotypes of IL2 promoter single nucleotide polymorphism -475 A/T as well as -631 G/A among individuals. X2 was calculated and Fisher’s exact test was applied to analyze the obtained data. The value of p<0.05 was considered significantly.Results: Evaluation of the -475 IL2 revealed that T allele and A/T genotype are present in 2% and 4% of MS patients, respectively, whereas T allele was absent in control samples. The comparison between alleles and genotypes in MS patients and healthy controls was not significant (p=0.1). For the -631 position, 1% and 2% of MS patients carried A allele and A/G heterozygote genotypes, respectively. All control samples had G allele and G/G genotype. The differences between patients and controls were not significant (p=0.4). Moreover, our results showed a very low frequency of T at -475 and A at -631 IL2 position in each of the two groups.Conclusion: Both -475 and -631 IL2 polymorphisms were higher in MS patients as compared to controls, but the frequency differences were not significant. Based on these data, it is suggested that the -475 and -631 IL2 polymorphisms as functional promoter position may be involved in IL2 expression and regulation. To find out the exact effect of the mentioned SNPs on susceptibility to MS, study on a larger sample size is suggested.

Background: Rheumatoid arthritis (RA) is a chronic autoimmune and inflammatory disease that affects the joints and consequently leads to the destruction of cartilage and bone lesions. Traditionally, ginger has been consumed in treatment of osteoarthritis, joint and muscle pain, neurological diseases, and inflammation of gums, tooth pain, asthma, stroke, diabetes, and constipation. The aim of this study was to determine the effect of ginger on some immunological and inflammatory markers in patients with rheumatoid arthritis. Methods: In this study, which was performed during 2013-2016, 66 patients with active rheumatoid arthritis who referred to the rheumatology clinic at Shariati hospital were en-rolled. Patients were randomly divided into 2 groups: one group consumed 1. 5 gr ginger per day, and the other group took roasted wheat flour (placebo), respectively. To determine the effect of confounding factors on the findings of the study, questionnaires for nutrient intake, physical activity, and medication were filled, and BMI was measured. For each participant, at the beginning and end of the study, Serum hs-CRP and mRNA levels of IL-1β , IL-2 and TNF-α were determined by ELISA and Quantitative Real Time PCR, respectively. Statistical analysis was performed using SPSS software. Significance level was set at p<0. 05. Results: Results of the study showed ginger powder supplementation caused a significant decline in CRP (p=0. 050) and IL-1β mRNA level (p=0. 021). TNFα mRNA levels reduced in ginger group compared to placebo groupalthough the difference was not significant between the 2 groups (p=0. 093). Ginger had no effects on IL2 gene expression. Conclusion: This study showed that ginger reduces inflammatory factors hs-CRP and IL-1β gene expression in patients with active RA and it seems that ginger can improve the inflam-mation in the patients.