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Showing 10 results for Delirezh

Kabiri F, Nejati V, Tukmechi A, Delirezh N, Nikbakhsh P,
Volume 68, Issue 12 (6 2011)
Abstract

Background: Lactobacillus species are genetically diverse groups of Lactic Acid Bacteria (LAB) that have been introduced as probiotics, because of some characteristics such as their anti-tumor properties, helping the intestinal flora balance, production of antibiotics, stimulation of host immune response, etc. The aim of this study was to investigate the effects of cytoplasmic extraction and cell wall of Lactobacillus species isolated from the intestine of common carp on human chronic myelocytic leukemia or K562 cancer cell lines.
Methods: The intestinal contents of 115 common carp captured from the natural resources of West Azerbaijan province in Iran were examined for LAB. After isolation, the identification of Lactobacilli was done according to traditional and molecular bacteriological tests. Subsequently, a suspension of each bacterium was prepared and the protein content of the cytoplasm was extracted. Cell wall disintegration was done by cell lysis buffer and sonication. The effects of cytoplasmic extraction and cell wall on K562 cell line proliferation were investigated by MTT assays.
Results: The cytoplasmic extraction of the isolated Lactobacilli had significant (p<0.05) anti-proliferative effects on K562 cells. The cytoplasmic extractions of Lactobacillus paracasei and Lactobacillus casei inhibited K562 cell proliferation by 66.56% and 54.28% at 83.33 μg/ml concentration, respectively.Nevertheless, the Lactobacillus cell wall could not inhibit the proliferations of K562 cells (p<0.05).
Conclusion: In this study, the cytoplasmic extractions of the isolated Lactobacilli from the intestine of common carp had anti-proliferative effects on K562cell line.


Asadi M, Farokhi F, Ganji Bakhsh M, Delirezh N, Nejati V, Gholami K,
Volume 69, Issue 1 (4 2011)
Abstract

Background: Nowadays, dendritic cells (DC) are used for tumor immunotherapy as they can induce immune responses against tumor cells. In this research, we comprehensively studied the maturation stimulus addition, PHA-activated T-cell (PHA- TCM) conditioned medium, autologous monocyte-conditioned medium (MCM) and TNF-α for their ability to promote uniformly mature dendritic cells that elicit T-cell responses. Methods: Plastic adherent monocytes were cultured with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) for five days and two days with monocyte-conditioned medium (MCM), tumor necrotizing factor-α (TNF-α) without TCM (PHA-activated T-cell conditioned medium). Phenotypic and functional analyses were carried out using anti-CD14, anti-CD80, anti-CD86, anti-CD83 monoclonal antibodies. Phagocytic activity, mixed lymphocyte reaction (MLR) and cytokine production were also evaluated. Results: The generated dendritic cells had high expression of surface molecules i.e. CD80, CD83, CD86 and HLA-DR. Moreover, the cells had low phagocytic and high T- lymphocyte stimulating activities. Measurement of the produced cytokines showed the generation of type-1 dendritic cells (DC1) in the study. Conclusion: The findings indicated that more efficient maturation of dendritic cells could be achieved by the use of PHA-activated T-lymphocyte conditioned medium in the culture medium. The aforesaid supernatant can be used as a maturation factor for the production of efficient dendritic cells with the ability to be used for tumor immunotherapy. This conditioned medium can provide new strategies and evolve into more advance tools for the generation of dendritic cells in vitro for tumor immunotherapy.
Gholami K, Nejati V, Delirezh N, Ganji Bakhsh M, Asadi M,
Volume 69, Issue 3 (5 2011)
Abstract

Background: The innate and adaptive immune responses are dependent on the migration of leukocytes across endothelial cells. Dendritic cells (DCs) play an important role in the initiation of cellular immune responses during their migration from tissues into the lymph nodes where they interact with endothelial cells of lymphatic vessels. We investigated the effects of surface-adherent and non-activated endothelial cells on phenotypic and functional characteristics of dendritic cells. Methods: Immature dendritic cells were generated from the isolation of peripheral blood mononuclear cells and their subsequent culture in DC-RPMI 1640 medium containing 10% FCS, interleukin-4 and granulocyte-macrophage colony-stimulating factor (GM-CSF) for five days. On day five, a maturation factor (composed of monocyte-conditioned medium, tumor necrosis factor-α (TNF-α) and poly I:C) was added to the RPMI medium where immature DCs were co-cultured with endothelial cell monolayer for 24 h. The maturation of harvested DCs on day seven was evaluated via flow cytometry, a beta-counter and an ELISA kit. Results: This study showed that the endothelial cells interact with dendritic cells generated from peripheral blood monocytes via cell-to-cell interaction. This interaction inhibits the maturation of DCs via decrease in the expression of CD83, CD86, CD80, HLA-DR and up-regulation of CD14. The interaction also inhibits the stimulation of T-lymphocytes resulting in a decrease in their proliferation. Conclusion: According to the findings of this study, it could be concluded that the endothelial cells can act as a potent regulator for DCs differentiation and function at the encounter made between them during the migration of DCs from tissues to lymph nodes.
Tanhaye Kalate Sabz F, Farokhi F, Delirezh N, Chapari H, ,
Volume 69, Issue 4 (6 2011)
Abstract

Background: Cell-therapy provides a promising alternative for the treatment of type 1 diabetes. Monocytes which have a reprogramming or differentiation potential and are more available than any other types of stem cells, have been recognized as candidates for such investigations. The aim of the present study was to evaluate the differentiation potential of rat peripheral blood monocytes into insulin-producing cells by the use of rat pancreatic extract (2 days after a 60% pancreatectomy).

Methods: Rat peripheral blood monocytes were isolated and cultured. Adherent monocytes were induced to differentiate into programmable cells in RPMI supplemented by 10% FCS, &beta-mercaptoetanol, M-CSF and IL-3 for six days. The dedifferentiated cells were analyzed by invert microscopy. Cultures of Programmable Cells of Monocytic Origin (PCMOs) were continued in RPMI, containing 10% FBS, pancreatic extract and 5 mmol/L glucose for 15 days. The medium was replaced every three days. At the end of the protocol, insulin and c-peptide excreted by the differentiated cells were tested by radioimmunoassay on days 6, 14, and 21. In order to verify insulin production in the cells, dithizone-staining, which is a method for insulin identification, was employed.

Results: The results showed that the cells cultured in rat pancreatic extract secreted insulin and c-peptide relative to the control group. Dithizone-staining was positive in the aforesaid cells (P<0/05).

Conclusion: The results of the current study showed that pancreatic extract treatment can differentiate rat peripheral blood monocytes into insulin-producing cells which can be regarded as a potential source for the treatment of diabetes.


Ganji Bakhsh M, Nejati V, Asadi M, Delirezh N, Farokhi F,
Volume 69, Issue 11 (4 2012)
Abstract

Background: Nowadays, dendritic cells (DCs) have a special place in cancer treatment strategies and they have been used for tumor immunotherapy as they can induce immune response against tumor cells. Researchers have been trying to generate efficient dendritic cells in vitro therefore, this research was done to generate them for use in research and tumor immunotherapy.

Methods: This study took place at Urmia University in 2010-2011 years. In this study plastic adherent monocytes were incubated with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) for five days. Finally, fully matured and stable DCs were generated by 48 hours of incubation in a monocyte conditioned medium (MCM) containing tumor necrosis factor-α (TNF-α) and epithelial cells. Phenotypic and functional analysis were carried out by using anti-CD14, anti-CD80, anti-CD86, and anti-CD83 monoclonal antibodies, and by determining their phagocytic activity, mixed lymphocyte reaction (MLR) and cytokine production, respectively.

Results: Dendritic cells were produced with high levels of surface molecule, i.e. of CD80, CD83, CD86, HLA-DR, expression and low levels of CD14 expression. Dendritic cells showed efficient phagocytosis and ability to stimulate T-lymphocytes. Moreover, dendritic cells could secrete high levels of interleukin-12 (IL-12) cytokine which was depictive of their full maturation. Measurement of the produced cytokines showed the generation of type-1 dendritic cells (DC1).

Conclusion: Our study showed that skin epithelial cells could induce maturation of monocyte-derived dendritic cells (DCs). This feeder layer led to the production of efficient dendritic cells with the ability to be used for tumor immunotherapy.


Chapari H, Farokhi F, Delirezh N, Javadi Sh, Tanhaye Kalate Sabz F,
Volume 69, Issue 11 (4 2012)
Abstract

Background: The characteristic of stem cells in self renewal and differentiation to different types of cells has stimulated the interests for using stem cells as a starting material for generating insulin secreting cells. We've evaluated the differentiation potential of Programmable cells of monocytic origin (PCMOs) into insulin producing cells effected from the growth factors and fibroblasts conditioned media (FCM).

Methods: Peripheral blood monocytes of rat were cultured for 6 days in RPMI with 15% FBS, β- mercaptoethanol, MCSF and interleukin-3. Then, these cells were incubated in differentiation media with HGF, EGF, Nicotinamide, 15% fibroblasts conditioned media and glucose for 15days. Morphological differences of cells were studied by invert microscope. In several stages, the amounts of insulin in supernatant of cells were measured by radioimmunoassay kit. Also productions of insulin from differentiated cells were studied with DTZ special staining.

Results: In response to MCSF and IL-3, monocytes dedifferentiated. These programmable cells of monocytic origin (PCMOs) were capable of differentiating into insulin producing cells in differentiation media. The morphology of differentiated cells was similar to Beta cells and the amount of insulin in supernatant of differentiated cells was much higher than PCMOs (P<0.05).

Conclusion: HGF, EGF, Nicotinamide and fibroblasts conditioned media are differentiation factors of PCMOs into insulin producing cells. According to the results insulin producing cells can be differentiated from programmable cells of monocytic origin in presence of fibroblasts conditioned media.


Abtahi Froushani Sm, Delirezh N, Hobbenaghi R, Mosayebi Gh,
Volume 69, Issue 11 (4 2012)
Abstract

Background: Recent studies have demonstrated an essential role for IL-17 in the pathogenesis of experimental autoimmune encephalomyelitis (EAE). Furthermore, it has been shown that FoxP3+Treg cells play an important role in the suppression of autoinflammatory reactions. Although, previous studies have determined the immunomodulatory potentials of all-trans-retinoic acid (ATRA), but these immunomodulations have been mostly justified by alteration in Th1/Th2 cytokines. The present study was carried out to investigate the therapeutic effects of ATRA on EAE and its effects on T-helper cells responses.

Methods: EAE was induced by MOG35-55 peptide and complete Freund's adjuvant in female C57BL/6 mice. The mice were allocated to two therapeutic groups (n=7 per group). Treatment with ATRA (500 μg/mouse every other day) was initiated in treatment group on day 12 when they developed a disability score. EAE controls received vehicle alone with the same schedule. Signs of disease were recorded daily until day 33 when the mice were sacrificed. Splenocytes were tested for proliferation by MTT test, cytokine production by ELISA and FoxP3+Treg cell frequency by flowcytometry.

Results: ATRA significantly reduced the clinical signs of established EAE. Aside from decreasing lymphocytic proliferation (P<0.05), ATRA significantly inhibited the production of pro-inflammatory IL-17 (P<0.005) as well as IFN-γ (P<0.0005) upon antigen-specific restimulation of splenocytes. FoxP3+Treg cell frequency and IL-10 levels were not altered significantly. However, IFN-γ to IL-10 and IL-17 to IL-10 ratios decreased significantly (P<0.0005).

Conclusion: Parallel to reducing autoreactive lymphocyte proliferation and cytokine production in favor of pro-inflammatory cytokines, all-trans-retinoic acid ameliorated established experimental autoimmune encephalomyelitis.


Loghmanni A, Delirezha N, Ownagh A, Mohebalianc H,
Volume 70, Issue 12 (5 2013)
Abstract

Background: Multiple Sclerosis (MS) is an autoimmune disease with impairment in function of central nervous system. Macrophages and dendritic cells play important roles in alleviating or progression of the disease. These cells can cause inflammation and damage to the myelin of nerve cells by realizing of harmful substances when these cells get matured. We studied the effect of Alternaria alternata extract on maturation of monocyte- derived dendritic cell (modc) and T-cell responses in the presence of Myelin Basic Protein (MBP) as a laboratory model of multiple sclerosis (MS). The purpose of this study is suitable dendritic cells production for usage in MS immunotherapy.
Methods: For this study plastic adherent monocytes were cultured with granulocyte/ macrophage- colony stimulating factor (GM-CSF) and interleukin -4 for converting these cells to modc and pulsed with MBP and matured in the presence of monocyte-conditioned medium (MCM) in control group and MCM + Alternaria alternata extract in treatment groups. Anti-CD14, anti-CD83, anti-human leukocyte antigen-DR (anti HLA-DR) monoclonal antibody were carried out for phenotyping. Autologos T cell responses and cytokine production were evaluated.
Results: The results showed that the expression of CD14 decreased and CD83, HLA-DR increased in treatment groups in comparison with control groups. The production amount of IL-10 overcame IL-12 and in T cell the production of cytokines, IL-17 and Interferon-γ (IFN-γ) decreased and IL-4 was increased (P<0.05). These effects escalated with increasing of dosage from 50 to 100 (mg/ml) (P<0.001).
Conclusion: Alternaria alternata extract can cause maturation of MBP-pulsed modc and skewing of T- lymphocyte toward Th2 and thereby can evolve into a new strategy in immunotherapy of MS.


Leyla Soleymani , Rahim Hobbenaghi , Aram Mokarizadeh , Samad Zare , Nowruz Delirezh ,
Volume 72, Issue 7 (October 2014)
Abstract

Background: Recently, bone-marrow-derived cells have introduced new therapeutic approaches to the management of wound healing in severe skin injuries. Bone marrow-derived stromal cells are described as a heterogeneous population, including mesenchymal stem cells, hematopoietic stem cells, and fibro-blast cells. Results derived from several studies indicate that these cells may contribute to tissue regeneration whether through producing variety of bioactive growth factors and/or by differentiation into mesoderm lineage. The aim of the present study was to investigate the effect of subcutaneous administration of bone marrow-derived stromal cells in repairing or regeneration of skin wounds induced by third-degree burn in a mouse model. Methods: In an experimental study that was performed in Urmia University research center from December 2011 to June 2012, The third-degree skin burn was induced on the shaved backs of healthy 7-8 week old male mice (N=18) using a metal rods heated in boiling water. After 1 hour, based on the equal physical condition mice were randomly divided into two separate groups and then subcutaneously administered with phosphate buffered saline (PBS 400 µl) or bone marrow-derived stromal cells (106 cell in 400µl PBS) at the burn site. 7, 14 and 21 days after induction of burn injury, biopsies were taken from burn wounds and then the sections were prepared. Subsequently the prepared sections were stained with hematoxylin/eosin and Masson's trichrome to explore histopathological changes evoke by administration of bone marrow derived stromal cells in comparison with control subjects. Results: Considering investigated parameters including formation of granulation tissue (respectively on days 7, 14 and 21 P≤ 0/007, P≤ 0/0013 and P≤ 0/001), angiogenesis (on day 21 P≤ 0/002) and collagen deposition, in mice treated with bone marrow-derived stromal cells the rate of healing of third-degree thermal burns was significantly accelerated when compared to the PBS-treated mice. Conclusion: This experimental modulation of wound healing suggests that bone marrow-derived stromal cells can significantly enhance the rate of wound healing possibly through stimulation of granulation tissue, angiogenesis, fibroblast proliferation and collagen deposition.
Reza Habibian , Nowruz Delirezh , Amir Abbas Farshid ,
Volume 73, Issue 5 (August 2015)
Abstract

Background: Allergic Asthma is an inflammatory disease of the respiratory system that is well known by increased inflammatory cells in the airways and causes difficulty in respiration. The prevalence of allergic asthma is increasing worldwide, and it has become a significant cause of health challenge especially in developed countries. Inhaled β2-agonists and Inhaled or oral corticosteroids are common medications for treating the disease, but they cannot be used for long periods of time because of frequently occurring side effects and they can’t change the main pathogenesis of the problem. Deficiency in regulatory system against inflammation could be an important factor in allergic asthma. Mesenchymal stem cells (MSCs) have potential of cellular immunosuppressive therapy of inflammatory disorders. The aim of present study was to evaluate the effects of MSC therapy on mechanisms of allergic asthma in mice model. Methods: This experimental study was conducted from August 2014 to March 2015. The animals were housed and maintained in Biotechnology Center of Urmia University, Iran. Mice were sensitized by intra-peritoneal injection of ovalbumin (OVA) and aluminum hydroxide emulsion and then were challenged intra-nasally with OVA. Before allergen challenge on day 14, experimental mice received tail vein injection of MSCs in PBS. Regulatory T cells of spleen, cytokines and IgE analysis were carried out using lungs wash as well as serum samples. Results: Our results showed that MSCs significantly reduced total cells and eosinophilia, serum OVA-specific IgE concentration in OVA-sensitized and challenged mice. Also results showed that MSCs markedly inhibited expressions of Th2 cytokines and elevated levels of Treg cells and Treg cytokines. Conclusion: In the present study, we demonstrated the inhibitory effect of MSCs on airway inflammation using mice model of allergic asthma. The mice were sensitized with OVA and compared to the results of dexamethasone administration. Our results demonstrated that administration of MSCs could be used as a potential therapeutic approach for the allergic asthma.

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