Background:The premature termination of high producer clones, which will be killed due to cell proliferation and proteins production antagonism, is one of the basic drawback in recombinant proteins technology. Furtheremore, it is supposed some toxic proteins like INTERFERON which we intended to clone and express, inhibit host cells’ proliferation. So, it is necessary to tightly control IFN-γ production during growing and selecting of highly producing clones. Methods: In the present study, we constructed an expression vector, pMPGB43P2(6)K containing the cumate-regulatable expression cassette to high production of human IFN-γ in Chinese Hamster Ovary- Cumate Transactivator (CHO-CTA). The clones were selected in the cumate and without cumate treated medium. Results: Our results showed, induced IFN-γ expression level was about 5 of magnitude higher than the constitive transgenic system. Conclusion: Application of cumate-regulatable expression cassette, which can be switch off and on by cumate, is useful to production of high producer clones and express toxic proteins in animal cells.      

INTERFERON gamma (IFN-g) is a cytokine produced by activated T lymphocytes and natural killer cells. It plays an important role in innate and adaptive immunity. IFN-g gene is polymorphic and its haplotype has been associated with resistance to nematode infestations which are the most important parasites of domestic sheep worldwide. To evaluate IFN-g polymorphism in Iranian Shaul sheep population, 136 blood samples were collected. Genomic DNA were extracted and amplified by PCR using specific primers for exon 3 of IFN-g gene. After gene amplification, SSCP and Heteroduplex mobility analysis carried out on polyacrylamid gel. Five unique SSCP patterns were detected in this population which 3 of them represent 3 alleles. Although heteroduplex mobility analysis demonstrated that all samples were homozygous, SSCP could reveal heterozygosity at frequency of 40. 4% in the studied population. Data obtained from the present study have revealed that IFN-g gene was a highly polymorphic in Iranian Shaul ecotype. This will be useful to study host-pathogen interaction and to evaluate association between resistance or susceptibility to disease with IFN-g in this ecotype.

Background: Not enough data are available about the effectiveness of consensus INTERFERON (CIFN) among HCV genotype 3 patients who failed to respond to pegylated INTERFERON and ribavirin.Objectives: We aimed to assess the efficacy and safety of CIFN and ribavirin in non-responders and relapsers to pegylated INTERFERON with ribavirin therapy.Patients and Methods: This open-label investigator-initiated study included 44 patients who received CIFN 15 mg /day plus ribavirin 800-1200 mg daily. In patients with an early virological response (EVR), the dose of CIFN was reduced to 15 mg thrice a week for further 36 weeks. Patients with delayed virological response continued to receive daily CIFN plus ribavirin to complete 48 weeks. The patients were considered “non-responders” if there were less than 2 log reduction in HCV RNA at 12 weeks and detectable HCV RNA at 24 weeks.Results: Twenty-four patients (55%) were non-responders and 20 patients were relapsers to the previous treatment with pegylated INTERFERON plus ribavirin (mean age 43.6 ± 9.4 years, males 25 (57%)). Nine patients were clinically cirrhotic (Child A). End of treatment virological response was achieved in 19 (43.1%) patients and sustained virological response (SVR) occurred in 12 (27.3%). Out of these 12 patients, eight were non-responders and four were relapsers to the previous treatment. Advanced fibrosis or clinical cirrhosis was associated with low SVR. Adverse events were fever, myalgia, anorexia, depression, and weight loss. Two patients received granulocyte colony stimulating factor for transient neutropenia. Seven patients were given erythropoietin to improve hemoglobin, and six were treated for mild depression. Two patients developed portosystemic encephalopathy.Conclusions: More than one-quarter of treatment-experienced patients with HCV genotype 3 achieved SVR after re-treatment with consensus INTERFERON plus ribavirin.

Objective Resistance exercise and supplementation by Shirazi thyme has beneficial effects on inflammatory processes. The present study aims to assess the effect of 8 weeks of resistance exercise combined with Shirazi thyme supplementation on plasma levels of INTERFERON-gamma (INF-γ, ) and Tumor Necrosis Factor-alpha (TNF-α, ) in men with Non-Alcoholic Fatty Liver Disease (NAFLD). Methods This is a clinical trial with a quasi-experimental design. Participants were 60 non-athlete men with NAFLD selected using a convenience sampling method and divided into four groups of placebo (n=15), exercise (n=15), supplementation (n=15), and exercise + supplementation (n=15). The exercise program consisted of eight weeks of resistance exercise at 3 sessions per week with an intensity of 50-80% of one repetition maximum). For supplementation, a 20 mg soft capsule (Barij Gastrolith) was prescribed three times per day and half an hour before meals. Blood samples were taken before and after the intervention. Data analysis was conducted in SPSS software using one-way analysis of variance considering a significance level of 0. 05. Results Resistance exercise (9. 20±, 5. 67), Shirazi thyme supplementation (8. 36±, 4. 80) and their combination (7. 80±, 4. 02) had no significant effect on INF-γ,level (p≥, 0. 05). Moreover, resistance exercise (4. 56±, 1. 69), Shirazi thyme supplementation (4. 25±, 2. 43) and their combination (5. 21±, 2. 22) had no significant effect on TNF-α,level (p≥, 0. 05). Conclusion Resistance exercise alone, Shirazi thyme supplementation alone, or their combination cannot improve inflammatory markers of fatty liver in people with NAFLD. However, it requires further research.

Hepatitis C Virus is one of the main reasons for chronic liver disease and hepatocellular carcinoma. Combination therapy with INTERFERON (peg-IFN-α) and Ribavirin (RBV) clear the virus more likely than the others. Different factors like virus and host characteristics influence on response to treatment. The most important viral factors include virus genotype and viral load, host factors like genetic, gender, race, age, weight and liver enzymes are also important. Previous studies have shown that single nucleotide polymorphisms (SNPs) in IFNR genes can regulate and influence on treatment with IFN. The purpose of this study is to investigate the association between SNPs in IFN-α receptor (IFNAR1 & IFNAR2) genes among subjects affected with chronic hepatitis C, who have treated with IFN and RBV, and also relationship between HCV genotypes and response to combination antiviral therapy. Peripheral blood mononuclear cells (PBMCs) were taken from whole blood of 61 patients affected with chronic hepatitis C who were treated with IFN and Ribavirin. Then, DNA was extracted from PBMCs and quality of DNA was assessed with Nanodrop finally two SNPs [Ex4-30G>C] and [Ivs1-4640 G>A] of IFN receptor genes (IFNAR1 and IFNAR2) were measured by TaqMan Real-Time PCR in ABi Prism 7900 system. Also to confirm the response rate to therapy, RNA was extracted then RT PCR was performed and final product was studied with gel electrophoresis and UV spectroscopy. Statistical analysis was performed using SPSS version 18.0 for Windows. The analysis of results from TaqMan SNP Genotyping has been shown that two SNPs (Ex4- 30G>C and Ivs1-4640 G>A) of IFNAR1 and IFNAR2 did not show any relationship with response to combined therapy in subjects affected with chronic hepatitis C who have treated with peg-IFN-α and Ribavirin.61 patients complete the treatment period.54 patients (%88/5) of them responded to treatment and 7 patients (%11/5) did not. Research and data analysis have shown that there is no significant relationship between sex (P=0.7) and age (P=0.2). But there is a relationship between genotype-3a and response to combined therapy of IFN-α and RBV (0.02). Studies have shown that gene polymorphisms in IVSS1-22G location of IFNAR1 gene had a relationship with IFN treatment response. But current study has shown that there is no significant relationship between two SNPs Ex4-30G>C and Ivs1-4640 G>A and response to IFN therapy. In continue we suggest that it would be better to use this technique to evaluate other SNPs in IFN genes.