The Effect of IFN-ß Treatment on Plasma Levels of BDNF and IL-6 in Relapsing-Remitting Multiple Sclerosis Patients.

BACKGROUND: In recent investigations addressing neurodegenerative diseases, especially multiple sclerosis (MS), the roles of brain-derived neurotrophic factor (BDNF) and interleukin-6 (IL-6) have been examined. METHODS: Forty-five relapsing-remitting MS (RRMS) patients, including 32 IFN-ß-treated and 13 newly identified untreated cases as well as 45 sex- and age-matched healthy controls, were recruited in the study. Plasma levels of BDNF and IL-6 were assessed using the ELISA method. Data were analyzed by SPSS (ver.21). RESULTS: There were significant differences between the case and healthy control groups in terms of the plasma levels of BDNF (p value = 0.044) and IL-6 (p value <0.001). Besides, the treatment with IFN-ß had no significant impact on the level of BDNF or IL-6 in RRMS patients as compared to healthy controls (p value = 0.716 and 0.623 for BDNF and IL-6, respectively). Furthermore, the increase in the plasma levels of BDNF and IL-6 indicated a direct correlation in the case group (r = 0.508, p value = 0.008). In detail, following the classification of the case group into 2 subgroups of IFN-ß-treated and untreated patients, a direct positive correlation was observed between the plasma levels of BDNF and IL-6 in IFN-ß-treated patients (r = 0.495, p value = 0.026). CONCLUSION: The IFN-ß treatment seems not to be effective for upregulating BDNF and IL-6 in RRMS patients.

Modulation of Th17 Proliferation and IL-17A Gene Expression by Acetylated Form of Apigenin in Patients with Multiple Sclerosis.

The presence of Th17 cells in CNS lesion of MS patients due to their inflammatory cytokines secretion is in line with the deterioration of the disease. Currently, the use of natural compounds with anti-inflammatory properties such as flavonoids have been considered to reduce inflammation in these patients, but the remaining issue is how deliver these compounds to the site of inflammation. Acetylation is a way to better uptake compound by cells and cross through cellular layers with tight junctions. This study aimed to investigate the in vitro effects of the Apigenin 3-Acetate on Th17 cells of MS patients and compare its efficacy with Apigenin and Methyl Prednisolone Acetate. IC50 for Apigenin 3-Acetate, and Methyl Prednisolone Acetate were determined using three healthy volunteers. The peripheral blood mononuclear cells (PBMCs) of five MS patients were isolated and co-cultured with a selected dose of Apigenin, Apigenin 3-Acetate, and Methyl Prednisolone Acetate for 48 hr, and then theproliferation of Th17 cells in isolated PBMCs was assessed by flow cytometry. The levels of RAR-related orphan receptor (RORC) and IL-17A expression were also determined by quantitative real-time PCR. The results showed that Apigenin 3-Acetate inhibited Th17 cells proliferation (P value: 0.018) at 80 µM concentration after 48 hr. Additionally, IL-17A gene expression significantly (P value≤ 0.0001) inhibited by Apigenin, Apigenin 3-Acetate and Methyl Prednisolone Acetate in 80 µM, 80 µM and 2.5 µM (selected dose in IC50 determination) respectively These results demonstrate that Acetate increases anti-inflammatory effects of Apigenin on Th17 cells.

Therapeutic effects of extracellular vesicles from human adipose-derived mesenchymal stem cells on chronic experimental autoimmune encephalomyelitis.

Since in cell therapy, there are always concerns about immune rejection, genetic disability, and malignancies, special attention has been paid to extracellular vesicles (EVs) which are secreted by mesenchymal stem cells (MSCs). In the present study, we assessed and compared the therapeutic effects of human adipose-derived mesenchymal stem cells (hADSC) and hADSC-EVs from adipose tissue on experimental autoimmune encephalomyelitis (EAE). After induction of EAE in C57Bl/6 mice, they were treated with hADSCs, hADSC-EVs, or vehicle intravenously. The clinical score of all mice was recorded every other day. Mice were killed at Day 30 and splenocytes were isolated for proliferation assay and determination of the frequency of Treg cells by flow cytometry. Leukocyte infiltration by hematoxylin and eosin, percentages of demyelination areas by luxol fast blue, and mean fluorescence intensity of oligodendrocyte transcription factor 2 (OLIG2) and myelin basic protein (MBP) by immunohistochemistry were assessed in the spinal cord. Our results showed that the maximum mean clinical score and myelin oligodendrocyte glycoprotein-induced proliferation of splenocytes in hADSC- and hADSC-EV-treated mice were significantly lower than the control mice (p < .05). We also demonstrated that the frequency of CD4+ CD25+ Foxp3+ cells was significantly higher in the spleen of hADSC-treated mice than EAE control mice (p = .023). The inflammation score and the percentages of demyelination areas in hADSC- and hADSC-EV-treated groups significantly declined compared with the untreated control group (p < .05). We also showed that there was no significant difference in MFI of MBP and OLIG2 in the spinal cord of studied groups. Overall, we suggest that intravenous administration of hADSC-EVs attenuates the induced EAE through diminishing proliferative potency of T cells, mean clinical score, leukocyte infiltration, and demyelination in a chronic model of multiple sclerosis.

Mesenchymal Stem Cell-Derived Extracellular Vesicles: A Novel Cell-Free Therapy.

In recent years, mesenchymal stem/stromal cells (MSCs) have widely been considered as therapeutic tools in basic researches and clinical trials. Accumulating evidence supports the idea that MSCs perform their therapeutic roles in paracrine manner especially through trophic factors and extracellular vesicles (EVs). Compared to cells, EVs have several advantages to be used as therapeutic agents, such as they lack self-replicating capabilities, dangers of ectopic differentiation, and tumor formation, genetic instability, and cellular rejection by the immune system. Since the MSC-derived EVs (MSC-EVs) appear to exert similar therapeutic effects of their parent cells, such as ability to arrive themselves to the site of injury and immunomodulatory properties, MSC-EVs have been widely studied in many animal models, including kidney, liver, cardiovascular, immunological, and neurological diseases. Regarding this, MSC-EVs look to be a novel and interesting approach to be studied in clinical trials of different inflammatory diseases. In this review, we summarize the properties and applications of MSC-EVs in different diseases.

Male microchimerism in peripheral blood from women with multiple sclerosis in Isfahan Province.

Multiple sclerosis (MS) is referred to as an organ-specific T-cell-mediated autoimmune disease of the central nervous system (CNS). Different genetic and environmental factors increase the risk of developing MS. In recent years, microchimerism (Mc) has been widely studied in autoimmune diseases, although the exact role of this phenomenon in human health is not known well. Microchimerism is the low level presence of DNA or cells from one individual into the tissue or circulation of another individual. In the current study, we evaluated the association of fetal microchimerism (FMc) with MS in Isfahan province. In this study, we enrolled 68 women in four groups. Two groups were MS patients with or without a pregnancy for a son, and the other two groups were MS-negative patients with or without a pregnancy for a son. The presence of the male genome assessed and compared in these groups. Four millilitres of peripheral blood were collected from all subjects in the tube containing EDTA and DNA was extracted. Real-time PCR assay was used for the DAZ (deleted in azoospermia) region Yq 11.23 as a marker for male microchimerism in all subjects. Our results showed that the percentage of DAZ (male genome)-positive women was significantly higher in MS-positive women given birth to a son in comparison with the other three groups. Our results also revealed no significant correlation between the percentage of DAZ-positive women and Expanded Disability Status Scale (EDSS) score and age of onset in the patients' group. For future studies, we suggest enrolling subjects who MS diagnosis occurred before and after pregnancy with a son. Comparing FMc in these two groups might provide a better understanding of the possible role of FMc in later development of MS.

Evaluation of plasma Osteopontin level in relapsing- remitting multiple sclerosis patients compared to healthy subjects in Isfahan Province.

Background: Multiple sclerosis (MS) is known as a neuroinflammatory disease of the central nervous system (CNS). The neuroinflammation may induce pro-inflammatory cytokines such as Osteopontin (OPN). OPN plays an important role in the inflammation by modulating the T helper1 (Th1) and Th17 responses. Since the exact immune pathogenesis of MS is complex and not well defined and many factors are involved, the need to detect more contributing biomarkers may help in setting new therapeutic strategies.Objective: This study tried to compare plasma OPN levels in relapsing- remitting multiple sclerosis (RRMS) patients during the remission phase with healthy subjects in Isfahan province.Materials and methods: In a case-control study, plasma was collected from the 40 RRMS as well as 38 (age and sex matched) healthy individuals as a control group. PlasmaOPN level was measured and compared between the two groups by Enzyme-linked immunosorbent assays (ELISA).Result: Statistical analysis revealed that plasma OPN level was markedly higher in the case group (RRMS patients during the remission phase) compared with the control group (P- value = 0.039). Our results also showed that there was no statistically significant difference in mean of plasma OPN level among RRMS patients who were treated with interferon (IFN)-ß and those who were not (P- value = 0.332). There was also no correlation between OPN plasma level and EDSS score (r = 0.037, P- value = 0.835), age of onset (r = 0.161, P- value = 0.357) and duration of disease (r = 0.121, P- value = 0.490).Conclusion: Higher OPN plasma level in studied patients suggests that OPN increased in RRMS patients who were in remission phase. It could be hypothesized that plasma OPN level may be increased as part of the pro-inflammatory cytokine milieu taking place in MS patients. OPN may not be specific marker for MS, but targeting it might present promising therapeutic effect to MS patients.

Negative Regulation of Semaphorin-3A Expression in Peripheral Blood Mononuclear Cells Using MicroRNA-497-5p.

BACKGROUND: Semaphorin-3A (Sema3A), as a secreted semaphorin, is an immune modulator molecule participating in the pathogenesis of autoimmune diseases. MicroRNAs (miRNAs) modulate the target-gene expression at the post-transcriptional level. It has been proposed that miRNAs may be crucial to the modulation of the function of semaphorins. Previous findings have proven that miR-497-5p is upregulated and Sema3A is downregulated in some autoimmune disorders. Thus, we aimed to examine the presence of any correlation between Sema3A and miR-497-5p in peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs were cultured and transfected with miR-497-5p mimic using the X-tremeGENE™ reagent. The expression level of Sema3A was assessed after 48 hours in supernatants and cells via the enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction, respectively. Cell viability was evaluated using the methylthiazol tetrazolium assay. All the experiments were done in triplicate, and the statistical analyses were performed with SPSS, version 20. P values equal to or less than 0.05 were considered significant. RESULTS: We observed downregulation of the Sema3A gene (P=0.0001) and its secretion (P=0.032) in the PBMCs through miR-497-5p transfection. Moreover, transfection with miR-497-5p mimic and downregulation of Sema3A did not affect the viability of the PBMCs (P=0.061). CONCLUSION: Based on the obtained results, we suggest that miR-497-5p has a high suppressive effect on Sema3A expression and both Sema3A and miR-497-5p can be considered critical targets for further studies on future therapeutic attempts for the treatment of autoimmune diseases such as multiple sclerosis and rheumatoid arthritis.

Serum level of interleukin 36 in patients with multiple sclerosis.

INTRODUCTION: Multiple sclerosis is a chronic autoimmune demyelinating disorder of central nervous system with unknown origin. In MS disease, T cells are pointed to myelin antigens and it leads to myelin loss and axonal degeneration. Cytokines are important regulators of immune system and has critical roles in MS pathogenesis. Interleukin 36, a member of interleukin 1 family, has been shown having important roles in some autoimmune disorders due to its proinflammatory actions and its role in host immunity. METHODS AND MATERIALS: In the current study, 49 relapsing remitting multiple sclerosis patients and 41 healthy individuals were recruited. IL36 measurement was performed using enzyme-linked immunosorbent assay technique. RESULTS: Mean age of RRMS patient and control group were 31.84 ± 6.89 and 34.27 ± 8.83 years, respectively. Serum level of IL36 were 61.91 ± 16.29 in MS patients and 42.26 ± 17.54 in healthy group (P < 0.001). CONCLUSION: in this study for the first time, significantly higher serum level of IL36 was determined in RRMS patients comparing healthy individuals. This data may suggest important roles of this cytokine in MS pathogenesis.

IL-27 plasma level in relapsing remitting multiple sclerosis subjects: The double-faced cytokine.

Interleukin-27 (IL-27) is a member of IL-6/IL-12 cytokine family which possesses pro- and anti-inflammatory properties and participates in the pathogenesis of various autoimmune diseases. In our case-control study, plasma was collected from healthy subjects as control group (n = 40) and patients with relapsing-remitting multiple sclerosis (RRMS) (n = 40), including new identified cases without treatment (n = 12) and those treated with Interferon beta (IFN-ß) (n = 28). The plasma level of IL-27 was assessed by ELISA method. Our results indicated that plasma level of IL-27 in MS patients increased significantly compared to the control subjects (P = 0.027). Furthermore, after parting case group into the two sub-groups, results revealed a significant difference of IL-27 plasma levels between control group and treated patients (P < 0.001), but not about that of between healthy subjects and untreated MS patients (P = 0.259). Also, mean levels of IL-27 in treated and untreated patients showed a significant difference (P = 0.007). These results demonstrate the possible modulation of IL-27 during autoimmune disease in human which may suggest the suppressive or therapeutic role of IL-27 on inflammatory diseases.

IL-23 plasma level measurement in relapsing remitting multiple sclerosis (RRMS) patients compared to healthy subjects.

BACKGROUND: Multiple sclerosis (MS) is a chronic neuroinflammatory disease with unknown etiology and variable clinical evolution. Interleukin-23 (IL-23), a member of the IL-12 cytokine family is a heterodimeric cytokine composed of the IL-12p40 subunit, and with a novel p19 subunit, its ability to enhance the expansion of T helper type 17 (Th17) cells indicates the responsibility for many of the inflammatory autoimmune responses. OBJECTIVE: The objective of the project is to measure IL-23 level in plasma of multiple sclerosis (MS) patients in comparison with healthy control subjects. METHODS: In a case-control study, plasma was collected from healthy subjects as control group (n = 40) and patients with relapsing remitting multiple sclerosis (RRMS) (n = 40). The plasma level of IL-23 was assessed by ELISA method. Statistical analysis was performed with SPSS (Ver. 16). RESULTS: Plasma level of IL-23 in MS patients was significantly increased compared to control subjects (p Value < 0.001). CONCLUSIONS: Our findings revealed the increased IL-23 level in patients' group. In conclusion, the inhibition of IL-23 might be a novel and promising therapeutic strategy, especially in the therapy of autoimmune inflammatory diseases. IL-23 plays a pivotal role in development of MS and might be a specific marker and therapeutic target for MS.

Antidepressants and the Risk of Psoriasis Induction: A Case-Control Study.

Background: Psoriasis (PSO) is a common chronic autoimmune skin disease with a significant psycho-socio-economic burden. Some antidepressants (ADs) such as fluoxetine and bupropion can induce or exacerbate PSO. This study aimed to investigate the correlation between ADs history before PSO onset, and the risk of PSO induction, in Isfahan province, Iran. Materials and Methods: In this case-control study, 80 patients with PSO were selected by non-probability sampling method, and 80 healthy individuals were selected using simple random sampling. They were interviewed and medical information was recorded. Chi-square, Mann-Whitney, and Kruskal-Wallis tests for dichotomous or categorical data, and independent-sample t test for continuous data were used. Statistical significance was taken as P ≤ 0.05. Results: In this case-control study, a total of 160 individuals, 80 participants in each group, were included. The mean age of the total samples was 44.8 ± 16 years. Forty-three percent of the individuals were women. PSO familial history in the cases was significantly higher than the control group (OR = 11.94, P = 0.001). It was revealed that use of ADs by patients before PSO induction, was greater than the controls (OR = 2.78, P = 0.058). Conclusions: Past history of ADs in the cases before PSO onset, was higher than the controls, indicating a possible association between ADs and the risk of PSO induction. This study can be effective to pay more attention to the possible complications of ADs and PSO risk factors. Accurate knowledge of PSO risk factors will be useful for better management and morbidity reduction.

The Immunomodulatory Aspect of Quercetin Penta Acetate on Th17 Cells Proliferation and Gene Expression in Multiple Sclerosis.

OBJECTIVE: The function of Th17 cells in the neuroinflammatory process in multiple sclerosis (MS) has been previously clarified. It has been suggested that Quercetin can influence MS due to a variety of anti-inflammatory effects. The present study aimed to examine in vitro immunomodulatory aspects of Quercetin Penta Acetate as a modified compound on Th17 cells of MS patients and also to compare its effects with Quercetin. MATERIALS AND METHODS: In this experimental study, peripheral blood mononuclear cell (PBMCs) were isolated and stained with CFSE then, half-maximal inhibitory concentration (IC50) values were determined using different doses and times for Quercetin Penta Acetate, and Methyl Prednisolone Acetate. Th17 cell proliferation was analyzed by flow cytometry and the expression levels of IL-17 and RORc genes were assessed by real-time polymerase chain reaction (PCR) method. RESULTS: The results showed that IL-17 gene expression was inhibited by Quercetin Penta Acetate (P=0.0081), but Quercetin Penta Acetate did not have a significant inhibitory effect on Th17 cells proliferation (P= 0.59) and RORc gene expression (P=0.1), compared to Quercetin. CONCLUSION: Taken together, our results showed some immunomodulatory aspects of Quercetin Penta Acetate on Th17 cells are more effective than Quercetin and it could be considered in the treatment of MS.

Blood IgMs from healthy donors and patients with systemic lupus erythematosus reduce the inflammatory properties of platelets from healthy donors.

Uncontrolled inflammation is the underlining mechanism of many human diseases and the increasing prevalence of such diseases mandate to develop new anti-inflammatory treatments. Utilizing the anti-inflammatory properties as well as other protective/beneficial features of natural IgMs (nIgMs) for treatment of human disorders seems as an easily accessible goal by the use of blood-purified IgMs as an alternative for polyclonal nIgMs. Despite the other blood cells, the functions of platelets have not been inspected under the influence of blood IgMs adequately. However, platelets, the second most numerous blood cells, are involved in the pathology of many inflammatory disorders through the production/expression of many inflammatory molecules. Thus, in the present study, we purified IgMs from serum of healthy donors and plasma of patients with systemic lupus erythematosus (SLE). Subsequently, we carried out comparative analysis of the inflammatory functions of normal platelets (P-selectin expression, GPIIb/IIIa activation, and secretion of soluble CD40L and TNF-α) that were stimulated by SLE microparticles (as key endogenous inflammation-drivers) in the presence or absence of the two IgM preparations; one with normal level of nIgMs (healthy blood IgMs) and the other with likely altered nIgM content (SLE blood IgMs). Both blood IgM preparations could suppress the elevated activation parameters of platelets in response to SLE microparticles. Additionally, the impact of SLE blood IgMs on the platelets was not superior to that of normal blood IgMs. The anti-inflammatory effects of blood IgMs on the activated platelets have been shown for the first time in the present study. Thus, this study provides evidence in favor of use of healthy blood IgMs as an anti-inflammatory therapy in clinical settings.

High-resolution melting curve analysis of polymorphisms within CD58, CD226, HLA-G genes and association with multiple sclerosis susceptibility in a subset of Iranian population: a case-control study.

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system with unknown etiology, which typically is manifested in early to middle adulthood. Recently, genome-wide association studies have identified susceptibility of immune-related genes to be involved in MS predisposition. The goal of the current study was to investigate the association of single nucleotide polymorphisms (SNP) with the immunologically related genes responsible for the disease, composed of CD58 (rs2300747 A>G), CD226 (rs763361 C>T), and HLA-G (rs1611715 A>C), with MS susceptibility. In this case-control study, a total of 200 patients suffering from relapsing-remitting multiple sclerosis and 200 healthy individuals were recruited. DNA was extracted from blood and then all subjects were genotyped for the polymorphism within mentioned genes by high-resolution melting (HRM) real-time PCR method. Statistical analyses were performed using SPSS software (version 20; SPSS, Chicago, IL, USA). Our finding showed that there are significant differences in genotype and allele frequencies between two groups regarding rs763361 (P = 0.035, OR 0.64, CI 95% for C allele) and rs1611715 (P = 0.038, OR 1.57, CI 95% for AA genotype) polymorphisms within CD226 and HLA-G genes, respectively. Concerning rs2300747 polymorphism on CD58 gene, no significant differences were found between cases and controls. In general, results from the current study indicate that CD226 and HLA-G, but not CD58 genetic polymorphisms are associated with increased risk of MS in Isfahan population similar to European populations. However, to elucidate how these SNPs contribute to MS pathogenesis, functional studies are needed.

Interleukin 18 Polymorphisms and its serum level in Patients with Multiple Sclerosis.

BACKGROUND: Multiple sclerosis (MS) is a chronic demyelinating disorder of central nervous system. Although the definite pathogenesis of MS has not been understood, crucial role of environmental and genetic risk factors has been proposed. PROPOSE: To determine the serum level of interleukin-18 (IL-18) as well as gene polymorphisms of IL-18 (rs1946518, rs360719, and rs187238). METHODS: In this case-control study, 110 MS patients diagnosed according to the McDonald criteria and 110 healthy individuals were recruited. IL-18 gene polymorphisms were genotyped by polymerase chain reaction high-resolution melt test, and IL-18 serum level was determined by enzyme-linked immunosorbent assay technique. RESULTS: The mean age of the MS patients (89 females and 21 males) and the control group (89 females and 21 males) was 30.3 ± 9.25 and 30.28 ± 9.13 years, respectively. The mean serum levels of IL-18 in MS patients and healthy individuals were 341.56 ± 39.22 Pg/Ml and 146.52 ± 29.30 Pg/Ml, respectively (P < 0.001). The genotype of rs1946518 (but not rs360719 and rs187238) was significantly different between groups (P = 0.037 and P = 0.069, respectively). CONCLUSION: In this study, we showed the significant higher IL-18 serum level and significant different frequencies of two polymorphisms of IL-18 in MS patients. These results show the important roles of IL-18 in MS pathogenesis. However, more studies are needed to verify our results in larger sample size.

Cell Density Counts of the Intestinal Intraepithelial Lymphocytes in the Celiac Patients.

BACKGROUND: Increased number of intestinal intraepithelial lymphocytes (IELs) is a key histological finding in the diagnosis of celiac disease (CD); however, the number of IELs in celiac patients and healthy subjects may vary from one region to another. Additionally, there are some seronegative celiac patients with a borderline histology. OBJECTIVE: To determine the number of the CD3+ and CD8+ IELs T-cells in the celiac patients and healthy subjects (controls) in Isfahan. METHODS: The duodenal biopsies were obtained from the celiac patients (n=15) and the controls (n=19). The total number of IELs/100 epithelial cells (ECs) were counted using the hematoxylin-eosin (H&E) staining method, and that of CD3+ and CD8+ IELs/100 ECs were counted using the immunohistochemistry (IHC) staining method. RESULTS: This study defined the upper normal limit for each variable as mean + 2SD. Accordingly, the upper normal limits of the total IELs, CD3+ IELs, and CD8+ IELs/100 ECs were calculated as 37 (95% confidence intervals, CI: 33-41), 22 (95% CI: 19-25) and 12 (95% CI: 10-14), respectively. In 3 clinically CD diagnoses, the total IELs counts/100 ECs were below the upper normal limit, and the histopathological and serologic assays were negative. Nevertheless, the CD8+ IELs T-cells counts/100 ECs showed borderline values. Interestingly, these patients responded to a gluten-free diet (GFD). CONCLUSIONS: The study findings suggest that in the clinically diagnosed celiac disease, IELs count/100 ECs below the upper normal limit as well as negative histopathological and serologic assays and the cell density counts of the CD8+ IELs T-cells/100 ECs could be a useful parameter in CD diagnosis and make a decision to put them on a GFD.

Immunoregulatory Effects of Silymarin on Proliferation and Activation of Th1 Cells Isolated from Newly Diagnosed and IFN-ß1b-Treated MS Patients.

Multiple sclerosis (MS) is a central nervous system autoimmune disease characterized by demyelination. Autoreactive T cells mainly interferon gamma (IFN-γ) producing T helper cells (Th1) have an important role in MS pathogenesis. Silymarin is a unique blend produced from milk thistle (Silybum marianum) plant which its imunomodulatory role has been indicated in studies. In the present study, the effects of silymarin on isolated Th1 cells were investigated in newly diagnosed MS patients and those who received betaferon. PBMCs were separated from newly diagnosed and IFN-ß-treated MS patients. The Th1 cell isolation from PBMCs was performed using a human Th1 cell isolation kit. Th1 cells were cultured in the presence of silymarin (50, 100, and 150 µM for 48, 72, and 120 h). Th1 cell proliferation and CD69 expression were assessed by flow cytometry. Also, IFN-γ production and T-bet gene expression were measured by ELISA and real-time PCR respectively. In vitro cultured Th1 cells showed that silymarin suppresses Th1 cell proliferation dose and time dependently in newly diagnosed and IFN-ß-treated MS patients in comparison to DMSO control. Also, CD69 expression as an early activation marker was changed after Th1 cell treatment with different doses of silymarin at different times. T-bet gene expression was significantly decreased in Th1 cells isolated from newly diagnosed and IFN-ß-treated RRMS patients after treatment with silymarin compared to DMSO control. Additionally, IFN-γ production by Th1 cells was decreased after treatment silymarin in newly diagnosed patients; however, in IFN-ß treated after 48-h treatment with silymarin, IFN-γ concentration was decreased at concentrations of 100 and 150 µM, and after 120 h, a significant increase was observed in the IFN-γ level at a concentration of 100 µM in comparison with DMSO. Our findings here clearly show that silymarin is an effective regulator for Th1 response in vitro condition. It not only suppresses Th1 proliferating activity but also inhibits T-bet gene expression and IFN-γ production by these cells.

Serum levels of histamine and diamine oxidase in multiple sclerosis.

BACKGROUND: Multiple Sclerosis (MS) is an autoimmune disease of the central nervous system, mostly affecting young adults. Diamine oxidase is an enzyme essential for histamine production. Histamine which is produced mostly by mast cells can have effects on different aspects of immune system via its different histamine receptors (H1R, H2R, H3R and H4R). The crucial role of diamine oxidase and histamine in immune balance has been documented in different studies and experiments both on MS patients and on experimental allergic encephalomyelitis (EAE). In this regard, we aimed to measure the level of histamine and diamine oxidase in the serum of MS patients. METHODS: A total number of 50 relapsing-remitting MS (RRMS) patients and 41 age and sex matched controls were enrolled in this study. Assessments of serum levels of histamine and diamine oxidase enzyme were performed using enzyme-linked immune sorbent assay (ELISA). RESULTS: The serum levels of histamine and diamine oxidase in RRMS patients were lower than healthy controls (P-value = 0.00, for both). CONCLUSION: Our research team found significant low levels of histamine and diamine oxidase in RRMS patients; however the pathogenesis of this issue was unclear.

Nanofibrous Tubular Membrane for Blood Hemodialysis.

As the most important components of a hemodialysis device, nanofibrous membranes enjoy high interconnected porosity and specific surface area as well as excellect permeability. In this study, a tubular nanofibrous membrane of polysulfone nanofibers was produced via electrospinning method to remove urea and creatinine from urine and blood serums of dialysis patients. Nanofibrous membranes were electrospun at a concentration of 11.5 wt% of polysulfone (PS) and dimethylformamide (DMF)/tetrahydrofuran (THF) with a ratio of 70/30. The effects of the rotational speed of collectors, electrospinning duration, and inner diameter of the tubular nanofibrous membrane on the urea and creatinine removal efficiency of the tubular membrane were investigated through the hemodialysis simulation experiments. It was found that the tubular membrane with an inner diameter of 3 mm elecrospun at shorter duration with lower collecting speed had the highest urea and creatinine removal efficiency. The hemodialysis simulation experiment showed that the urea and creatinine removal efficiency of the tubular membrane with a diameter of 3 mm were 90.4 and 100%, respectively. Also, three patients' blood serums were tested with the nanofibrous membrane. The results showed that the creatinine and urea removal rates were 93.2 and 90.3%, respectively.