Mohsen Mazloomrezaei; Mahsa Sadat Hosseini; Nahid Ahmadi; Elham Mahmoudi Maymand; Ebrahim Eftekhar; Amir Asgari; Amin Ramezani
Abstract
Background: It is advantageous to develop an effective purification procedure to produce recombinant protein drugs (rPDs) without any tags. To remove N- or C-terminus tags from the rPDs, several cleavage site-based endopeptidases were used. Separating the endopeptidase enzyme from the rPDs is a time-consuming ...
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Background: It is advantageous to develop an effective purification procedure to produce recombinant protein drugs (rPDs) without any tags. To remove N- or C-terminus tags from the rPDs, several cleavage site-based endopeptidases were used. Separating the endopeptidase enzyme from the rPDs is a time-consuming and costly process. Objective: To design and develop a new method for the purification of human interleukin (IL)-4 with potential application for other cytokines. Methods: Met-like amino acids were substituted at position 120 to reduce the possibility of alteration in the structure of IL-4 and its biological activity. Based on the in silico analysis, isoleucine was chosen as an alternative amino acid, and the M120I mutant IL-4 (mIL-4) model was selected for the downstream analysis. Recombinant mIL-4 was produced in the E.coli BL21 host and purified with CNBr. Then in vitro evaluations of the native and mutant IL-4 were performed. Results: The results showed that both the native and mutant IL-4 had the same effect on TF-1 cell proliferation. On the other hand, there was no significant difference between the effects of native IL-4 (nIL-4) and mIL-4 on the expression of IL-4 and IL-10 in activated peripheral blood mononuclear cells. Native and mutant IL-4 have similar biological activities. Conclusion: Here, an efficient and straightforward system is introduced to purify IL-4 cytokine using CNBr, which could be applied to other rPDs.
Xiaoling Liu; Xiaojia Liu; Yu Ren; Hongxin Yang; Xiaolei Sun; Haiyun Huang
Abstract
Background: Vitamin D supplementation has been proven to be effective in the treatment of allergic rhinitis (AR). Objective: We conducted the present study to explore the role and efficacy of vitamin D adjuvant therapy for the treatment of inflammation in patients with AR. Methods: Out of 127 patients ...
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Background: Vitamin D supplementation has been proven to be effective in the treatment of allergic rhinitis (AR). Objective: We conducted the present study to explore the role and efficacy of vitamin D adjuvant therapy for the treatment of inflammation in patients with AR. Methods: Out of 127 patients with potential eligible AR, 60 were randomly assigned into two groups and were finally included in our analysis (n=30 for each intervention). The patients with potential eligible AR were randomly allocated to intervention with desloratadine citrate disodium (DCD, 8.8 mg/day) without and with vitamin D3 nasal drops (1.5х106 IU, once/week) for four weeks. Thirty healthy control subjects were included in our study. We assessed the changes in the serum 25(OH)D, peripheral blood eosinophils, interleukin (IL)-4 levels, and nasal symptoms. Serum 25(OH)D, peripheral blood eosinophils, and IL-4 levels were detected respectively with liquid chromatography-tandem mass spectrometry (LC-MS/MS), a blood detector, and enzyme-linked immunosorbent assay. Results: Our patients who received vitamin D3 adjuvant therapy had a higher serum 25(OH)D level (47.57 ± 2.83 vs. 31.51 ± 2.95 ng/ml, p=0.000) and lower AR symptoms score (2.07 ± 1.89 vs. 3.37 ± 1.50, p=0.005), serum IL-4 (10.38 ± 3.41 vs. 12.79 ± 5.40 pg/ml, p=0.043), and peripheral blood eosinophils (0.34 ± 0.09 vs. 0.41 ± 0.10 109/l, p=0.003) compared with DCD single treatment. The efficacy rates of DCD with and without vitamin D3 in AR were 97% and 84%, respectively. Conclusion: Nasal vitamin D3 combined with DCD could improve the clinical symptoms of AR. Vitamin D3 adjunct therapy showed significant effects on inhibiting inflammation in patients with AR. We concluded that vitamin D3 supplementation could be an effective adjuvant therapy in AR patients.
Farhana Shahzad; Shahzad Tawwab; Nadeem Afzal
Volume 7, Issue 2 , June 2010, , Pages 109-116
Abstract
Background: Atherosclerosis is an inflammatory and multifactorial disease, with a high prevalence rate in Pakistan. Objective: To find a relation between serum IL-4 and IgE levels with oxidized LDL in atherosclerosis. Methods: In this observational, cross sectional study 99 male patients, between forty ...
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Background: Atherosclerosis is an inflammatory and multifactorial disease, with a high prevalence rate in Pakistan. Objective: To find a relation between serum IL-4 and IgE levels with oxidized LDL in atherosclerosis. Methods: In this observational, cross sectional study 99 male patients, between forty and sixty years of age, with a history of ischemic heart disease (IHD) and established atherosclerotic plaques on angiography were recruited. The study was completed within three years (Jan 2007 to Jan 2009). One hundred and one age and gender matched healthy subjects with no known history of IHD were also recruited. All the study participants were non-diabetics. Serum IL-4, IgE and oxidized LDL (ox-LDL) levels were measured by quantitative ELISA technique. Results: Serum IL-4 levels were generally undetectable or very low, but were higher in the patient group compared to the control subjects. Similarly, oxidized LDL and serum IgE levels were also increased in the patient group compared to the control, but the differences were not statistically significant. Conclusion: Our study could not detect any relationship between IL-4 and IgE levels with LDL oxidation in atherosclerosis.
