Enhancement the antioxidative and immunomodulatory functions of mesenchymal stem cells by tetrandrine.

In this study, mesenchymal stem cells (MSCs) were primed with Tetrandrine (TET) having anti-inflammatory and immunomodulatory effects to examine the effects of this molecule on the antioxidative potential of MSCs as well as their modulatory effects on activated peripheral blood mononuclear cells (PBMCs). The viability of primed MSCs was detected using MTT assay and Trypan blue staining. Moreover, flow cytometry technique was applied to evaluate cell cycle distribution and immunophenotype of MSCs. The production of superoxide dismutase 3 (SOD3), malondialdehyde (MDA), kynurenine, TGF-ß, and IFN-γ were also measured by spectrophotometry to assess the alteration of antioxidative and immunomodulatory potential of MSCs. Then, TET-primed MSCs were cocultured with PBMCs. The MTT assay was used to measure the proliferation of PBMCs. Cell cycle progression of PBMCs and frequency of regulatory T cells were evaluated using Flow cytometry. ELISA assay was also applied to determine the concentrations of TGF-ß and IFN-γ after coculturing. According to our data, TET enhanced the secretion of SOD3 and kynurenine from MSCs, while the production of IFN-γ was reduced. No changes were observed in the viability, proliferation, and immunophenotype of MSCs after priming with TET. Moreover, the proliferation and frequency of PBMCs in the S and G2/M phases of cell cycle reduced after co-culturing with TET-primed MSCs. The concentration of TGF-ß was increased in the supernatant of PBMCs, but the level of IFN-γ was reduced. Our data suggested this priming method as a novel strategy for increasing the antioxidative and immunomodulatory activity of MSCs.

Silibinin improved the function of T cells in peripheral blood mononuclear cells (PBMCs) co-cultured with U-87 MG cell line.

Objective: Silibinin has exhibited antitumor activities. However, there are few reports about the immunomodulatory properties of silibinin on T lymphocyte function in the tumor microenvironment. Here, we determined the effects of silibinin on T cells of peripheral blood mononuclear cells (PBMCs), cultivated alone or with a human cell line of glioblastoma (U-87 MG). Materials and Methods: The proliferation of T lymphocytes was assessed by MTT test in the presence of silibinin (15 and 45 µM). Also, total antioxidant capacity (TAC), the activity of superoxide dismutase-3 (SOD3), and the levels of two cytokines interferon gamma (IFN-γ) and tumor growth beta (TGF-ß) were compared between treated and untreated PBMCs alone or co-cultured with U-87 cells. Results: According to our results, silibinin raised the TAC levels and SOD3 activity in the PBMCs and in the co-culture condition. Moreover, silibinin-treated PBMCs showed higher IFN-γ levels and lower TGF-ß levels. Interestingly, silibinin protected PBMCs against the U-87-induced suppression. Conclusion: Altogether, these results proposed the immunomodulatory potential of silibinin on T cells of PBMCs, as well as its partially protective effects on PBMCs against the suppression induced by U-87 MG cells.

Metabolic Regulation of T cell Activity: Implications for Metabolic-Based T-cell Therapies for Cancer

Immunometabolism is an emerging field in tumor immunotherapy. Understanding the metabolic competition for access to the limited nutrients between tumor cells and immune cells can reveal the complexity of the tumor microenvironment and help develop new therapeutic approaches for cancer. Recent studies have focused on modifying the function of immune cells by manipulating their metabolic pathways. Besides, identifying metabolic events, which affect the function of immune cells leads to new therapeutic opportunities for treatment of inflammatory diseases and immune-related conditions. According to the literature, metabolic pathway such as glycolysis, tricarboxylic acid cycle, and fatty acid metabolism, significantly influence the survival, proliferation, activation, and function of immune cells and thus regulate immune responses. In this paper, we reviewed the role of metabolic processes and major signaling pathways involving in T-cell regulation and T-cell responses against tumor cells. Moreover, we summarized the new therapeutics suggested to enhance anti-tumor activity of T cells through manipulating metabolic pathways.

Protective potential of piroxicam on human peripheral blood mononuclear cells against the suppressive capacity of glioblastoma cell lines.

Dexamethasone, a common medication used in the treatment regimen of glioblastoma, has broad inhibitory effects on the immune responses. Here, in an in vitro study, we examined the effects of piroxicam, a potent substitute for dexamethasone, on peripheral blood mononuclear cells (PBMCs) co-cultured with two glioblastoma cell lines, U-87 MG and A-172 cells. MTT assay was used to determine the proliferation of PBMCs treated with piroxicam, or dexamethasone. In addition, to evaluate the effects of drugs on the cell cycle distribution, DNA content per cell was analyzed in PBMCs and A-172 cell lines using flow cytometry. Oxidative parameters, including superoxide dismutase-3 (SOD3) activity and total anti-antioxidant capacity, lactate dehydrogenase (LDH) activity, as well as IFN-γ and TGF-ß levels were measured in PBMCs alone or in the presence of cell lines using ELISA. Unlike dexamethasone, piroxicam showed a protective effect on PBMCs against both glioblastoma cell lines. Furthermore, while dexamethasone reduced the proliferation of PBMCs, piroxicam had no adverse effect on the proliferation. Cell cycle analysis showed a reduction in the G2/M phase in piroxicam-treated A-172 cells. Additionally, dexamethasone limited the cell cycle progression by increasing the fraction of PBMCs in G0/G1. Interestingly, after co-culturing piroxicam-treated PBMCs with cell lines, a remarkable rise in the LDH activity was observed. Although not significant, piroxicam partially decreased TGF-ß levels in both cell lines. Our findings suggested a protective effect of piroxicam, but not dexamethasone, on PBMCs against inhibitory mechanisms of two glioblastoma cell lines, U-87 and A-172 cells.

Effect of turmeric extract on bone healing in an experimental model of femoral bone fracture.

Objective: Following bone trauma, several factors participate in making a balance between the activity of osteoblasts and osteoclasts. The receptor activator of nuclear factor kappa B ligand (RANKL), receptor activator of nuclear factor kappa B (RANK), and osteoprotegerin (OPG) molecules play critical roles in the healing process via regulation of osteoclasts function. Turmeric is suggested to have an anti-osteogenic potential; however, its effect on accelerating bone healing has not been adequately studied. Here, we used a rat model of femur fracture to explore the effect of treatment with turmeric extract on the bone repair and the expression of RANK, RANKL, and OPG molecules. Materials and Methods: Eight rats were subjected to surgery, randomly divided into two groups, and treated orally with turmeric (200 mg/kg), or olive oil. Four oil-treated rats without bone fracture were used as control group. After six weeks of treatment, the femurs of animals were examined for radiological, histological, and gene expression analysis. Results: X-ray radiography showed thicker callus and a more obscure fracture line in the turmeric group. Furthermore, higher osteoblast percentages but no osteoclasts were observed in turmeric-treated animals, representing better repair of bone in the fracture site. Also, real-time analyses showed that treatment with turmeric reduced RANK and RANKL expression (p<0.0001) and lowered RANKL/OPG ratio (p=0.01) in femoral bone tissue. Conclusion: Our findings indicated the turmeric ability to facilitate bone hemostasis and optimize the expression of key markers involved in the bone metabolism.

Effect of Treatment with Thyme Extract on Urinary Levels of Melatonin in an Experimental Autoimmune Encephalomyelitis Mouse Model.

BACKGROUND: Thymus vulgaris, or thyme belongs to the Lamiaceae family of aromatic plant species and has established antioxidant and anti-inflammatory properties. We examined the association between thyme extract treatment to recovered urinary levels of melatonin, a hormone with neuroprotective effects, in mice induced with EAE. METHODS: Eight B6 mice induced with EAE were randomized into two groups and exposed to either 50 mg/kg of thyme extract or PBS. After EAE induction, mice were injected i.p every other day from day 0 to 21. Four B6 mice without EAE were considered the healthy control group. Urine samples were collected consecutively for two 24 h periods on day 19 and 20. We examined whether thyme extract treatment modified urinary melatonin sulfate concentration (ng/mL) in EAE-induced mice using an ELISA. RESULTS: The clinical score and body weight in thyme-treated EAE group were significantly lower in comparison to the EAE control group at indicated time points. The urinary melatonin concentration was significantly lower in the EAE control group compared to the healthy mice. There was no significant difference between thyme-treated and EAE groups regarding the urine melatonin concentration. CONCLUSION: Our results show that exposing EAE mice to thyme extract improved their clinical symptoms, however, there was no significant effect on urinary melatonin concentration.

Interaction Between Air Pollutants and Pollen Grains: The Role on the Rising Trend in Allergy.

Asthma and allergic diseases cases have risen in recent decades. Plant pollen is considered as the main aeroallergen causing allergic reactions. According to available data, urban residents experience more respiratory allergies than rural residents mainly due to the interaction between chemical air pollutants and pollen grains.This interaction can occur through several mechanisms; chemical pollutants might facilitate pollen allergen release, act as adjuvants to stimulate IgE-mediated responses, modify allergenic potential, and enhance the expression of some allergens in pollen grains. This review focuses on the most recent theories explaining how air pollutants can interact with pollen grains and allergens.

Impact of traffic-related air pollution on the expression of Platanus orientalis pollen allergens.

Air pollutants and their interaction with environmental allergens have been considered as an important reason for the recent increase in the prevalence of allergic diseases. The aim of this study was to investigate the traffic pollution effect, as a stressor, on Platanus orientalis pollen allergens messenger RNA (mRNA) and protein expression. P. orientalis pollen grains were collected along main streets of heavy traffic and from unpolluted sites in Mashhad city, in northeast Iran. The pollen samples were examined by scanning electron microscopy. To assess the abundance of pollen allergens (Pla or 1, Pla or 2, and Pla or 3) from polluted and unpolluted sites, immunoblotting was performed. Moreover, the sequences encoding P. orientalis allergens were amplified using real-time PCR. Scanning electron microscopy showed a number of particles of 150-550 nm on the surface of pollen from polluted sites. Also, protein and gene expression levels of Pla or 1 and Pla or 3 were considerably greater in pollen samples from highly polluted areas than in pollen from unpolluted areas (p < 0.05). In contrast, no statically significant difference in Pla or 2 protein and mRNA expression level was found between samples from the two areas. We found greater expression of allergens involved in plant defense mechanisms (Pla or 1 and Pla or 3) in polluted sites than in unpolluted ones. The high expression of these proteins can lead to an increase in the prevalence of allergic diseases. These findings suggest the necessity of supporting public policies aimed at controlling traffic pollution to improve air quality and prevent the subsequent clinical outcomes and new cases of asthma.

Quantification of Pla or 3, a Platanus orientalis Allergen, Grown under Different Environmental Conditions, by Sandwich ELISA.

BACKGROUND: Platanus species are widely cultured around the world and considered an important cause of allergic reactions. In the present study, we developed a sandwich ELISA to quantify Pla or 3 allergen in P. orientalis pollen extracts grown near high-traffic roads and compared it to pollen extracts collected from rural areas as control. METHODS: Pollen samples were collected from three polluted and two unpolluted sites in Mashhad, northeast Iran. Recombinant Pla or 3 was expressed and used for polyclonal antibody production in rabbit. A sandwich ELISA was developed and validated to quantify Pla or 3 levels in pollen extracts from the different sites. RESULTS: The coefficients of variation (CVs) for the intra- and inter- day assays were less than 5 and 18%, respectively. The working range of the standard curve was between 0.1 and 25 ng/ml, with the detection limit being 0.037 ng/ml. The recovery percentage was 88-106.4% at working concentrations from 0.31 to 26.5 ng/ml. Pla or 3 levels were significantly greater in pollens grown near high-traffic roads than in those grown in rural regions (p < 0.0001). CONCLUSION: A sandwich ELISA was developed and validated to quantify Pla or 3 in pollen extracts. Using this validated ELISA, we showed a substantial difference between the amounts of Pla or 3 in pollens grown in different environments. This finding should be considered in developing public policies to reduce traffic pollution, which leads to reduced allergic reactions in atopic subjects.

Alteration in mononuclear cell subpopulations in dogs immunized with gentamicin-attenuated Leishmania infantum.

The impact of immunization with gentamicin-attenuated Leishmania infantum (H-line) on the immunophenotypic profile of popliteal lymph node (PLN) and peripheral blood mononuclear cells (PBMCs) of dogs was assessed by flow cytometry and immunohistochemistry. Compared with the dogs infected with L. infantum wild-type (Group WT), there was a significantly higher percentage of CD4+, CD44+ T cells and CD14+, MHC-II+ cells and a lower percentage of CD4+ CD25+ regulatory T cells in PLN of the immunized dogs with L. infantum H-line (Group H). The percentage of CD4+ and CD8+ T cells in PBMCs of immunized dogs was higher than that in dogs of Group WT. The CD4:CD8 ratio in PLN of dogs of Group H was significantly higher than that in dogs of Group WT. A significantly higher percentage of CD21+ B cells and a lower percentage of CD79b+ cells were found in PLN of the immunized dogs compared with dogs of Group WT. Immunohistochemical investigation showed no parasites in the PLN of immunized dogs whereas there were parasites in the PLN of 60% of dogs infected with L. infantum WT. In this study, the immunophenotypic profile of mononuclear cells of the immunized dogs correlates with cellular immunity.