The effect of Rat mesenchymal stem cells and its soluble factors on peripheral blood neutrophil function

Background & aim: Mesenchymal stem cells (MSCs) are a population of adult stem cells which is an appropriate source for therapeutic purposes. The aim of this study was to investigate the effects of rat mesenchymal cells and soluble factors on the function of peripheral blood neutrophils. Methods:This experimental study was conducted on rat mesenchymal stem cells. Mesenchymal cells obtained from bone marrow of the femur and Tybaof 6-8 week rats and were cultured in DMEM. After maturation, the mesenchymal cells and supernatant at ratios of 1:4, 1:2 and 3:4 were adjacent with peripheral blood neutrophil phagocytosis. Subsequently, the respiratory burst of neutrophils, the yeast phagocytosis and nitroblue tetrazolium test was evaluated for revival. The Data were analyzed by t-tests, ANOVA and Tukey's test (p ˂0.05). Results:The rates of phagocyted neutrophil treated with MSCs compared to controls were decreased.This reduction was not statistically significant (p >0.05).The phagocitic cell in the rats of the treated group with supernatant compared to the control group in all three ratios of 1:4, 1:2, 3:4 increased significantly (p>0.05). by the increase in the ratio was observed (P>0.05). Respiratory burst of neutrophils treated with mesenchymal stem cells compared to the control group significantly decreased. Respiratory burst was increased in the groups treated with cell supernatant at ratios of 1:2 only (P>0.05). Conclusion:Mesenchymal cell-cell interaction with neutrophils was remarkable for therapeutic strategies in diseases associated with neutrophil function in response to physiological and pathological cell therapy with MSCs

Changes in some pro-and anti-inflammatory cytokines produced by bovine peripheral blood mononuclear cells following foot and mouth disease vaccination

Interleukin (IL)-17 is exclusively produced by CD4 helper T-cells upon activation. It most often acts as a pro-inflammatory cytokine, which stimulates the release of pro-inflammatory cytokines IL-6, IL-8, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF). In this study, we studied the in-vitro IL-17 response to specific antigens and a variety of mitogens and compared the IL-17 response to IL-2, IL-4, IL-5, IL-6, IL-10, and IFN-γ responses. We used a foot and mouth disease (FMD) vaccine as specific antigens and mitogens (phytohemagglutinin [PHA], pokeweed mitogen [PWM], and concanavalin A [Con A]) to stimulate peripheral blood mononuclear cells (PBMCs) of vaccinated calves. Cell culture supernatant was harvested and analyzed for cytokines, using commercially available bovine ELISA kits. The mitogens induced a significant increase in IL-17 production. IL-17 was produced at high levels in response to the T cell-stimulated mitogens, PHA, and Con A, and at low levels in response to PWM mitogens. In contrast, level of the produced IL-17 cytokines in response to the FMDV antigens was lower as compared to those produced by mitogens. The FMDV antigens and mitogens significantly increased IL-17 production. There was not a correlation between IL-17 production and type-1 cytokine, IFN-γ, and IL-2, while there was a correlation between type-2 cytokine, IL-4, and IL-5 at either cytokine level produced by PBMCs stimulated by FMDV antigens. Moreover, there was an interaction between IL-17 and IL-6, that is, as IL-6 cytokine level elevated or diminished, IL-17 cytokine level increased or decreased, as well

The Immunotherapeutic Effects of Pentoxifylline in Type 1 Diabetic Mice and its Effects on Expressions of Peroxisome proliferator-activated receptor gamma (PPARγ) gene

Background & aim: Pentoxifylline is an immunomodulatory and anti-inflammatory agent which inhibits the production of proinflammatory cytokines. The purpose of this study was to evaluate the effect of pentoxifylline in the treatment of type 1 diabetes in mice and its effect on the expression of peroxisome proliferator- activated receptor gamma (PPARγ). Methods: After induction of diabetes in male C57BL/6 mice, they were treated with Pentoxifylline (100 mg/kg/day) for 21 days. Blood sugar levels were measured on days 0, 7, 14 and 21. Splenocytes were tested for cytokines production by ELISA. Further investigations on immune system changes in spleens were tested by semi-quantitative RT-PCR on PPARγ gene. Statistical data were analyzed using the Student t-test and ANOVA. Results: Treatment with pentoxifylline prevented the level of blood sugar in diabetic rats. Pentoxifylline treatment also significantly inhibited the production of proinflammatory cytokines IL-17 and IFN-γ, while cause increasing the anti-inflammatory cytokine IL- 10 and the expression of PPARγ gene in the spleen compared to the diabetic control group (p< 0.05). Conclusion: Due to STZ induction, Pentoxifylline may have therapeutic effects against autoimmune destruction of pancreatic beta cells on type 1 diabetes in mic

Therapeutic effects of all-trans retinoic acid on experimental autoimmune encephalomyelitis and its role in T-helper lymphocyte responses

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

In-vitro differentiation of rat peripheral blood monocytes into insulin-producing cells by rat pancreatic extract

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, &amp;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&amp;lt;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

Inhibitiory properties of cytoplasmic extract of Lactobacilli isolated from common carp intestine on human chronic myelocytic leukemia K562 cell line: an in vitro study

&quot;n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 st1&quot;:*{behavior:url(#ieooui) } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} 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.&quot;n&quot;nMethods: 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.&quot;n&quot;nResults: The cytoplasmic extraction of the isolated Lactobacilli had significant (p&amp;lt;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 &amp;mu;g/ml concentration, respectively. Nevertheless, the Lactobacillus cell wall could not inhibit the proliferations of K562 cells (p&amp;lt;0.05).&quot;n&quot;nConclusion: In this study, the cytoplasmic extractions of the isolated Lactobacilli from the intestine of common carp had anti-proliferative effects on K562 cell line

Inducing maturation of monocyte-derived dendritic cells on human epithelial cell feeder layer

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