فهرست مطالب eisa salehi

Mechanisms underlying the systemic lupus erythematosus (SLE) have not yet been elucidated. In this study, we evaluated the balance of T cell subsets in BALB/c mice model of SLE induced; using Con A and polyamines as DNA immunogenicity modifiers. BALB/c mice were immunized subcutaneously with 50 µg extracted DNA from cells cultured in different conditions: splenocytes+ polyamines (group P), splenocytes+ Con A (group A), splenocytes+ polyamines+ Con A (group PA) and splenocytes only (control). Anti-double-stranded DNA –(ds-DNA) antibodies, proteinuria, and antinuclear autoantibodies were assessed by enzyme-linked immunosorbent assay, Bradford method, and immunofluorescence respectively. Transcription factors of different T helper subsets were examined by real-time polymerase chain reaction. The serum level of the anti-dsDNA antibody in group PA was higher than that in the other groups (p>0.05). Antinuclear antibody (ANA) titer increased in groups A and PA. Proteinuria level in group PA was significantly higher than that in the control group (p<0.001). Expression of Foxp3 was decreased in group A (p=0.001). Additionally, the ratios of T-bet/GATA3 and T-bet/Foxp3 were also increased in group A. (p>0.05). Our results revealed an increased ratio of Th1 to Th2 and decreased expression of Foxp3 in group A, but group PA manifested more obvious signs of the disease. These results suggest that other mechanisms rather than disturbance in T cells' balance may involve the development of disease symptoms.

It is estimated that 10% of people receiving hepatitis B (HB) vaccination fail to produce a protective level of hepatitis B surface antibody (HBsAb). Various methods such as administration of immune-enhancing medications, increasing the dose of vaccine, or changing injection route are implemented to overcome this issue.
In this regard, to the current study aimed at assessing the efficacy of atorvastatin treatment in subjects not responding to HB vaccination and evaluating the possible molecular pathways included in the process.
In the current clinical trial, healthy subjects with HBsAb titers of less than 10 IU/mL after a complete course of HB vaccination were included. Participants were randomly assigned into two groups of case and control. Subjects in the case group received daily atorvastatin (40 mg) tablets for 10 days, while the controls were given identical placebo tablets. On the 5th day all subjects received 1.0 mL intramuscular HB vaccine. Four to eight weeks after vaccination, blood samples were drawn from the participants for laboratory assessments including enzyme-linked immunosorbent assay (ELISA) to measure the level of HBsAb, interleukin (IL)-4, IL-17, interferon (IFN)-γ and transforming growth factor (TGF)-β cytokines, and real-time polymerase chain reaction (PCR) was also used to evaluate the expression of their corresponding genes; T-Bet, RORc, and GATA3.
A total of 30 subjects (five females and 25 males) were included, with 15 participants in each group. There was no significant difference between the two groups considering baseline characteristics of the participants. Final laboratory assessment of HBsAb levels revealed that 12 (80%) subjects from the case and 5 (33.3%) from the control groups produced a protective level of antibodies (P = 0.025). No significant difference was observed in the concentration of IL-4, IL-17, IFN-γ, and TGF-β, and the expression of their corresponding genes between the two groups.
The study showed that short-term atorvastatin administration was associated with an increased level of HBsAb to that of protective levels against HB, but this response was not induced through the assessed immune pathways. Further investigations are required to identify the exact mechanism for this effect of atorvastatin.

Preeclampsia is one of the most common complications of pregnancy that occurs after the 20th weeks of pregnancy. The pathophysiology of this disease is not exactly known. Transforming Growth Factor-Beta (TGF-β) and Nitric Oxide (NO) are the key regulatory factors secreted by Mesenchymal Stem Cells (MSCs). The aim of the present study was to evaluate the TGF-β and NO secretion by adipose-derived MSCs in normal and preeclamptic pregnant women.

The adipose tissues were collected from 10 preeclamptic patients and 10 age-matched normotensive controls at the time of cesarean section delivery. After isolation and expansion of MSCs, their capability of differentiation and immunophenotyping characteristics were assessed. Next, the release of TGF-ß was evaluated making use of ELISA sandwich method and Griess method was used to measure the level of NO.

Adipose derived MSCs in both groups were differentiated into osteocytes and adipocytes. The expression of CD90, CD73, CD44, and CD105 markers and lack of expression of CD-14, CD34, CD45, and HLA-DR markers in cells isolated from adipose in both groups was observed using flow cytometric analysis. The levels of TGF-ß secretion in preeclamptic women were significantly higher than those in control group, but the mean level of NO secreted by adipose derived MSCs did not significantly change in the two groups.

It can be concluded that significant increase in the secretion of TGF-β owing to MSCs in preeclamptic participants shows the importance of these cells in controlling immunological balance in these patients. Therefore, MSCs-based therapy seems to regulate TH1/Th2 balance in preeclampsia.

Preeclampsia is one of the most common complications of pregnancy that occurs after the 20th weeks of pregnancy. The pathophysiology of this disease is not exactly known. Transforming Growth Factor-Beta (TGF-β) and Nitric Oxide (NO) are the key regulatory factors secreted by Mesenchymal Stem Cells (MSCs). The aim of the present study was to evaluate the TGF-β and NO secretion by adipose-derived MSCs in normal and preeclamptic pregnant women. The adipose tissues were collected from 10 preeclamptic patients and 10 age-matched normotensive controls at the time of cesarean section delivery. After isolation and expansion of MSCs, their capability of differentiation and immunophenotyping characteristics were assessed. Next, the release of TGF-ß was evaluated making use of ELISA sandwich method and Griess method was used to measure the level of NO. Adipose derived MSCs in both groups were differentiated into osteocytes and adipocytes. The expression of CD90, CD73, CD44, and CD105 markers and lack of expression of CD-14, CD34, CD45, and HLA-DR markers in cells isolated from adipose in both groups was observed using flow cytometric analysis. The levels of TGF-ß secretion in preeclamptic women were significantly higher than those in control group, but the mean level of NO secreted by adipose derived MSCs did not significantly change in the two groups. It can be concluded that significant increase in the secretion of TGF-β owing to MSCs in preeclamptic participants shows the importance of these cells in controlling immunological balance in these patients. Therefore, MSCs-based therapy seems to regulate TH1/Th2 balance in preeclampsia.

Regulatory T cells (Tregs) are important components of the immune system that modulate responses of other cells. These cells are involved in peripheral tolerance mechanisms, so defect in development and function of these cells can result in autoimmune disease. Increasing evidence supports the role of microRNAs-21 (miR-21) in the regulation of forkhead box P3 (Foxp3) expression in Tregs. We aimed to determine whether miR-21 transfection to naive CD4 T cells can be useful in generation of iTregs in-vitro. We investigated in-vitro differentiation of miR-21-transfected naive CD4 T cells to iTregs and compared these iTregs to cytokine-differentiated iTregs and control group. We showed that expression of Foxp3, transforming growth factor beta (TGF-β), and interleukin-10 (IL-10) are increased in iTregs generated after miR-21 transfection in comparison with cytokine-differentiated iTregs and control group. Our findings demonstrate that miR-21 has positive role in in-vitro generation of induced regulatory T-cells (iTregs).

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease which results in damage to various organs. Some animal studies have revealed that activation of Toll-like receptors (TLRs) is important in the pathogenesis of SLE. In the present study, the percentage of different immune cell subsets in 35 SLE patients and 38 control subjects was analyzed by flow cytometry. We also assessed the expression of TLR9 in the population of peripheral blood mononuclear cells (PBMCs) including T lymphocytes (CD4 and CD8), B lymphocytes (CD19), NK cells (CD56) and monocytes (CD14) in SLE patients and healthy controls. The results showed that the percentage of CD8 T lymphocytes and CD14 monocytes were significantly higher (p˂0.001) in the SLE patients than the healthy control subjects. Moreover, the percentage of CD56 NK cells were significantly lower in the SLE patients than the healthy control subjects (p=0.001). The findings indicated that the expression of TLR9 was significantly higher in CD4 and CD8 T lymphocytes and CD19 B lymphocytes of SLE patients than in control subjects (all p˂0.05). The difference in TLR9 expression are involved in pathogenesis of the SLE, hence it can be used as an indicator for SLE diagnosis.

An appropriate differentiation of distinct human CD4+ T cell subset is critical for manipulating these cells for using in immunity related diseases. Despite various attempts to clarify the role of different factors involved in Th17 differentiation, many crucial contradictions yet remained to be optimized. Although it has been shown that the differentiation of in-vitro Th17 cells culture conditions requires the presence of IL-1beta, IL-23, IL-2, IL-21, IL-6 and TGF-β, the optimum amount of TGF-β regulating in vitro human Th17 cell differentiation is still unclear. In the current study, a flow cytometric assay was used to evaluate the effect of different concentrations of TGF-β and a combination of IL-1beta, IL-23, IL-2 without using IL-6 on development of Interleukin (IL)-17–producing T helper (Th17) cells. According to our findings, 0.1 ng/ml of TGF-β significantly increases the expression of IL-17 in comparison to other concentrations of this cytokine. Results indicated the vital role of TGF-β cytokine in the polarization of human Th17 cells in vitro.

Recent studies have shown the immunomodulatory effect of vitamin D3 through down-regulation of Toll-like receptor (TLR) expression in human monocytes. In this study, the effects of vitamin D treatment on TLR2 and TLR4 expression on monocytes derived from type 2 diabetes was investigated.
To assess the influence of vitamin D3 on expression of TLR2 and TLR4 on monocytes from patients with type II diabetes, peripheral blood sample was taken of 30 patients. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifuge and then monocytes were isolated from these cells with using the magnetic activated cell sorting (MACS). To investigate the effect of vitamin D3 on the expression of TLR2 and TLR4, monocytes were cultured in the presence of vitamin D3 (10-9 M) for 48 hours. Then the expression of TLR2 and TLR4 was determined by Real-time PCR.
We found that vitamin D3 suppresses the mRNA expression of TLR2 and TLR4 in patients with type II diabetes. TLR2 and TLR4 expression in the patients exposed to vitamin D3 were significantly decreased in comparison with patients who were not treated with vitamin D3.
It can be concluded that vitamin D3 supplements may be further analyzed as a therapeutic option by reducing TLR2 and TLR4 expression in patients with type II diabetes.

Recurrent respiratory infections are one of the clinical protests of those with delayed complications of exposure to mustard gas. Studies Show that inflammatory mediators have been changed in serum, sputum and Broncho-Alveolar Lavage (BAL) samples of chemical victims. Regarding TLR4 importance in inflammation and response to infections, this study aimed to investigate TLR4 gene expression in chemical victim's lung tissues. In this case-control study, paraffin blocks of lung tissue samples were collected from 28 veterans exposed to mustard gas with delayed pulmonary complications (cases group) and 9 pulmonary samples from the subjects with no mustard exposure (controls group) from the archive of some hospitals during 2013-2014. Slides samples from lung paraffin blocks stained with Hematoxylin and Eosin (H&E) were prepared. Real-time PCR was used to evaluate RNA expression. ΔΔCT median was used to measure the relative genes expression. Comparison of data was done using Mann-Whitney test by SPSS 21 software. In case group, 12 patients (42.8%) were diagnosed with constructive bronchiolitis, 8 patients (28.6%) with respiratory bronchiolitis and 8 patients (28.6%) with other disease and in control group, 4 patients (44.4%) with constructive bronchiolitis, 1 patient (11.2%) with respiratory bronchiolitis and 4 patients (44.4%) were diagnosed with other diseases. Case group ΔΔCT median was 5.28±3.58 cyc and that of control group was 5.81±3.29cyc that gad no significant difference (p>0.05). As there is pathological similarity in lung tissues of both groups, it seems that TLR4 gene expression undergoes same changes.

Epidermal growth factor (EGF) is a polypeptide molecule, with important functions in epithelial growth and wound repair. It exerts its effects on cells by binding to receptors on the cell surface. The aim of this study was to evaluate and compare salivary EGF levels in patients with gingivitis and advanced periodontitis as well as in healthy controls. Unstimulated salivary samples were collected from patients with gingivitis and advanced periodontitis and healthy individuals. The clinical parameters of plaque index (PI), bleeding on probing (BOP), probing pocket depth (PPD) and clinical attachment level (CAL) were measured and recorded using a Williams probe. The enzyme-linked immunosorbent assay (ELISA) was used to determine salivary levels of EGF. One-way ANOVA was used for data analysis. The mean salivary level of EGF in healthy individuals (99.27) was significantly higher than that in patients with gingivitis (61.53). This value in patients with gingivitis (61.53) was also significantly higher than that in subjects with periodontitis (36.14) (P<0.001). The reduction in salivary level of EGF in patients with periodontal disease may be related to the pathogenesis of periodontitis.

Multiple Sclerosis (MS) is a chronic inflammatory disease that leads to degeneration of the brain and spinal tissue. Imbalances of CD4+ T cells including Thelper1 (Th1)/Thelper2 (Th2) and Thelper17 (Th17)/Tregulatory (Treg), their secreted cytokines and gene expressions, are important aspects of in immunopathogenesis of MS. Vitamin A and its metabolites can regulate the immune system and appears to be effective in preventing progression of the autoimmune disease such as MS. Disease progression was evaluated By Magnetic Resonance Imaging (MRI), Expanded Disability States Scale (EDSS) and Multiple Sclerosis Functional Composite (MSFC) tests. Cytokine levels were measured using ELISA kits and gene expression was quantified by Real time PCR (RT-PCR) system. According to the difference between the epidemiological and clinical data on the relationship between vitamin A and immune system regulation, this study of the first time assesses Immune function as well as gene expression and progression of the disease following administration of vitamin A supplement.

Interleukin (IL)-17-producing T helper (Th)-17 cells have recently been explained as a distinct population of CD4+ T cells which play an important role in immunity against infectious agents. Establishment of persistent phenotype of Th17 cells and recognition of lineage-deviating factors are of most attractive goals in modern researches in immunology. Although IL-6 and TGF-β are frequently used to differentiate naive T cells to Th17 phenotype in mouse models, the application of IL-23 and its importance in preventing cells from plasticity needs to be more investigated. Our main objective was to evaluate the role of IL-23 in Th17 to Th1 plasticity. In this research project, we generated in vitro Myelin oligodendrocyte glycoprotein (MOG)-specific Th17 cells in the presence of TGF-β, IL-6, IL-23 and peptide MOG35-55. Th17 development was confirmed by assessment of relevant transcription factors and secreted cytokines by flowcytometry and ELISA, respectively. Th17 to Th1 plasticity was monitored by consecutive samplings in different time points without any extra supplementation of IL-23. Cell culture supernatant was evaluated for Interferon (IFN)-γ secretion and cells were evaluated for intracellular expression of this cytokine. Our results showed that the employed method was relatively convenient in developing antigen-specific Th17 cells. We also showed that IL-23 deprivation which happens by prolongation of culture period, can convert IL-17 producing cells to IFN-γ secreting Th1 phenotype. IL-23 can be considered as a Th17 phenotype stabilizing factor for in-vitro developed lineages.

Preeclampsia is one of the most common complications during pregnancy that occurs after 20th weeks of pregnancy in women with normal blood pressure. The pathophysiology of this disease is unknown. Due to changes in serum level of nitric oxide in women with preeclampsia and also the important role of mesenchymal stem cells in the secretion of nitric oxide as an immunoregulator، in this study، we aimed to evaluate the level of nitric oxide in secretions of adipose tissue-derived mesenchymal stem cells in women with normal pregnancy and patients with preeclampsia. Subcutaneous adipose tissue of 10 preeclamptic patients and 10 healthy pregnant women was collected during cesarean operation. After isolation and proliferation of mesenchymal stem cells، capablity of their differentiation and their immunophenotyping charactetistics were assessed. Then، their release of nitric oxide was evaluated using Griess method. Stem cells isolated from adipose tissue in both groups differentiated into osteocyte and adipocytes. Flow-cytometric analysis showed the expression of the markers CD90، CD73 CD44 and CD105 and lack of expression of the markers CD-14، CD34، CD45 and HLA-DR in both groups. No significant change was observed for the level of nitric oxide secretion in both groups. The present results suggest that nitric oxide secreted by mesenchymal stem cells do not have a significant contribution in variation of serum level of this factor and possibly do not play a role in the pathology of preeclampsia. However، such study with a higher sample size is suggested.

Allergic rhinitis (AR) is an inflammatory disorder of the nasal mucosa with high morbidity and prevalence. Natural killer (NK) cells might have a role in AR. We aimed to evaluate the changes of the markers and receptors on NK cells in AR patients compared to the non-atopic controls. Flow cytometric analysis was used with double staining of the Peripheral Blood Mononuclear Cells (PBMCs) to examine the expression of CD25 and CD69 markers, and NKG2D and NKG2A receptors on NK cells of 20 patients with AR and 20 non-atopic controls. The serum total IgE level was measured by Enzyme-linked Immunosorbent Assay. The expression of CD69 antigen on NK cells in AR patients was significantly higher than that of healthy group (p=0.03). No significant changes were observed between CD25, NKG2D and NKG2A expression on the surface of NK cells from healthy and AR subjects. Our study also showed that there was no significant correlation between the expression of CD69, CD25, NKG2D and NKG2A and level of serum total IgE in AR patients and normal subjects. These results indicated that the expression of CD69 antigen on NK cells of AR patients was increased. The high expression of CD69 on NK cells in AR patients suggested that these cells were activated, probably due to the cytokines secreted from allergen-stimulated T cells and activated monocytes.

Atherosclerosis is a chronic inflammatory condition that affects the arterial wall. Oxidized lowdensity lipoprotein (ox-LDL) seems to have an important role in atherosclerotic plaque formation. This study was performed to investigate the effects of ox-LDL as well as PHA on proliferation and gene expression of peripheral blood mononuclear cells (PBMCs) in patients with atherosclerosis compared to healthy controls. Proliferation of PBMCs was assessed by BrdU assay, while gene expression was assessed by real-time PCR. Both PHA and ox-LDL significantly induced proliferation of PBMCs of patients and controls. PBMCs from controls showed significantly higher proliferation when stimulated with ox-LDL compared to patients. Expression of TGF-. was significantly lower in PBMCs from patients compared to healthy controls (p<0.001). Following simulation with PHA, TGF-. and Foxp3 gene expression levels in patients and controls were significantly decreased (p<0.001). Expression of Foxp in PBMCs treated with ox-LDL was significantly decreased in patients and controls. Decreased expression of TGF-. and Foxp3 genes after ox-LDL stimulation may be due to more sensitivity of Treg cells than effector T cells to ox-LDL. Presence of ox-LDL within atheroma could be associated with the diminished population of Treg cells in the atherosclerotic patients.

All-trans retinoic acid (ATRA), as an active metabolite of vitamin A, has been shown to affect immune cells. This study was performed to evaluate the effect of ATRA on viability, proliferation, activation and lineage-specific transcription factors of CD4+ T cells. CD4+ T cells were separated from heparinized blood of healthy donors and were cultured in conditions, some with, some without ATRA. Viability was assessed by PI flowcytometry and proliferation was measured by MTT assay. CD69 expression was determined by flowcytometry as a measure of cell activation. Lineage-specific transcription factors (FOXP3, RORγt and T-bet) were examined by intracellular staining and flowcytometry. High doses of ATRA (0.1-1 mM) caused extensive cell death in both PBMCs and CD4+ T cells. Doses of ATRA equal to or lower than 10 µM did not adversely affect cell viability and proliferation in comparison to culture medium without ATRA. Doses of ATRA between 10 µM and 1nM significantly increased cell activation when compared to culture medium without ATRA. ATRA could increase FOXP3+ and also FOXP3+RORγt+ T cells while it decreased RORγt+ and T-bet+ T cells. This study showed that doses of ATRA up to 10 µM are safe when using with CD4+ T cells in terms of cell viability, proliferation and activation. We could also show that ATRA diverts the human immune response in neutral conditions (without adding polarizing cytokines) by increasing FOXP3+ cells and decreasing RORγt+ cells. ATRA could be regarded as a potential therapy in inflammatory conditions and autoimmunities.

Although current treatment options for cervical cancer rely on platinum-based chemoradiotherapy, individualized approaches to therapy may improve response or reduce unnecessary toxicity. Overexpression of Excision repair cross-complementing 1 (ERCC1) has been associated with Cisplatin resistance in some tumors. We hypothesized that ERCC1 overexpression is related to treatment response. 32 patients with cervical cancer were enrolled. Malignant tissue was isolated from pretreatment biopsies, and quantitative real-time reverse transcriptase polymerase chain reaction assays were performed to determine ERCC1 expression. Patients were divided to ERCC1 positive and ERCC1 negative. Response to chemoradiotherapy was evaluated and compared among the two groups. The mean age of participants was 56.6±12 years. Objective response was obtained in 24 patients (75%). ERCC1 was 2.8 times higher in patients who did not respond to treatment compared with the responders (OR: 2.8). ‍ Assessment of ERCC1 expression in tumoral tissue is possible in the clinical setting and predicts response to chemoradiotherapy. Further studies are necessary for final judgment.

Allergic rhinitis is a common disorder with great morbidity. Its prevalence has increased during recent years, therefore attracting attentions to its mechanisms. Type 2 cytokines play a major role in allergies. It has been proposed that Natural killer (NK) cells may be able to produce type 2 cytokines. This study was done to evaluate NK cells number and subtypes in patients with allergic rhinitis, comparing healthy subjects. >>>

Apart from genetic and environmental factors, activation of autoreactive mechanisms has been proposed to play a role in the pathogenesis of schizophrenia. In re-cent years, considerable work has been carried out to understand the role and contribution of the immune system in this disease. To investigate the T cell response to phytohaemagglutinin (PHA) and determine the serum levels of anti-nuclear antibody (ANA), anti-cytoplasmic antibody (ACA), and circulating immune complexes (CIC) in schizophrenic patients. A total of 30 drug-free schizophrenic patients and 42 healthy controls were enrolled in this study. T cell proliferation in response to PHA was measured using Methyl Thiazol Tetrazolium test. ANA and ACA were measured by indi-rect immunofluorescence. CIC concentration was determined using poly ethylene glycol precipitation assay. Mean PHA response was 1.96 ± 0.83 in patients and 3.72±1.39 in healthy controls (p < 0.001). ANA and CIC concentrations were not signifi-cantly different between two groups. In addition, ACA was detected only in patients. Increased production of ACA together with lower T cell response to mito-gens in our patients provides evidence for the involvement of autoimmune mechanisms in the pathogenesis of schizophrenia.

Schizophrenia has been associated with altered immunity. Different studies regarding natural killer cell activity (NKA) in schizophrenic patients have shown inconsistent results. To evaluate NK cell activity in schizophrenic patients in comparison with healthy control individuals. 30 medication-free schizophrenic patients and 41 healthy sex، age and smoking status matched individuals were included in this study. NK cell activity of case and control subjects was measured by Methyl-Thiazol-Tetrazolium (MTT) test. Statistical analysis of the data was done using SPSS 11. 5 software. NK activity of patients and normal subjects had a mean of 36. 94 ± 26. 15 (Mean ± SD) and 22. 31 ± 17. 92، respectively. A significant increase in NK activity in schizophrenic patients compared to controls (P = 0. 011). Among patients، NK activity of smokers was significantly lower than that of non-smokers (P = 0. 02). Other demographic factors didn''t show any influence on NK activity. The higher activity of NK cells in the schizophrenic patients as compared with the control population could explain the low incidence of cancer in these patients. Decreasing the effect of smoking on NK activity in the patients could be one of the responsible factors for the inconsistency in the results of different studies.